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4,334,902,168,058,104,300 | Anti-GSK3B Rabbit Polyclonal Antibody [clone: 9HCLC]
Supplier: Pierce Biotechnology
710100
CAPI710100EA 1361.64 CAD
CAPI710100
Anti-GSK3B Rabbit Polyclonal Antibody [clone: 9HCLC]
This antibody is predicted to react with non-human primate, mouse and rat based on sequence homology. ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A. ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production. Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain. Glycogen synthase kinase-3 beta (GSK3 beta) belongs to the subfamily of glycogen synthase kinases, that play a central role in energy metabolism, neuronal cell development, and body pattern formation. GSK3 has alpha and beta isoforms (51 kDa and 47 kDa respectively), that are functionally dissimilar. GSK3 beta plays a role in insulin and Wnt signaling, and is inhibited following the extracellular signaling events.
Recommended Dilutions: Western Blot: 1:500-1:5000, Immunofluorescence: 1:100-1:1000, Immunocytochemistry: 1:100-1:1000, ChIP assay: 1 ?l
Type: Primary
Antigen: GSK3B
Clonality: Polyclonal
Clone: 9HCLC
Conjugation: Unconjugated
Epitope:
Host: Rabbit
Isotype: IgG
Reactivity: Human
Order Now
Learn more
About VWR
Avantor is a vertically integrated, global supplier of discovery-to-delivery solutions for...
Learn more About VWR | {
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-1,063,808,952,737,339,600 | Original source
Variants (including SNPs and indels) imported from dbSNP (release 138) | View in dbSNP
Alleles
T/C | Ancestral: T | Ambiguity code: Y | MAF: 0.39 (C)
Location
Chromosome 3:156798775 (forward strand) | View in location tab
Most severe consequence
Evidence status
Synonyms
Archive dbSNP rs17385692
HGVS name
3:g.156798775T>C
Genotyping chips
This variation has assays on: Illumina_ExomeChip | {
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-1,471,385,055,951,995,000 | About Help FAQ
Smad4 Gene Detail
Summary
• Symbol
Smad4
• Name
SMAD family member 4
• Synonyms
D18Wsu70e, Dpc4, DPC4, Madh4, Smad 4
• Feature Type
protein coding gene
• IDs
MGI:894293
NCBI Gene: 17128
Location & Maps
more
• Sequence Map
Chr18:73639009-73703780 bp, - strand
• From VEGA annotation of GRCm38
Mouse Genome Browser
• Download
Sequence
64772 bp ± kb flank
• Genome Browsers
• Genetic Map
Chromosome 18, 49.51 cM
• Mapping Data
10 experiments
Homology
more
• Human Ortholog
SMAD4, SMAD family member 4
• Vertebrate Orthologs
9
• Human Ortholog
SMAD4, SMAD family member 4
Orthology source: HomoloGene, HGNC
• Synonyms
DPC4, JIP, MADH4, MYHRS
• Links
NCBI Gene ID: 4089
neXtProt AC: NX_Q13485
UniProt: Q13485
• Chr Location
18q21.2; chr18:51030213-51085042 (+) GRCh38.p7
Human Diseases
more
• Diseases
1 with Smad4 mouse models; 2 with human SMAD4 associations
Human Disease Mouse Models
IDs
View 1 model
IDs
Click on a disease name to see all genes associated with that disease.
• Mutations/Alleles
2 with disease annotations
• References
4 with disease annotations
Mutations,
Alleles, and
Phenotypes
less
• Phenotype Summary
153 phenotypes from 19 alleles in 28 genetic backgrounds
54 phenotypes from multigenic genotypes
8 images
180 phenotype references
Phenotype Overview
adipose tissue
behavior/neurological
cardiovascular system
cellular
craniofacial
digestive/alimentary system
embryo
endocrine/exocrine glands
growth/size/body
hearing/vestibular/ear
hematopoietic system
homeostasis/metabolism
integument
immune system
limbs/digits/tail
liver/biliary system
mortality/aging
muscle
nervous system
pigmentation
renal/urinary system
reproductive system
respiratory system
skeleton
taste/olfaction
neoplasm
vision/eye
Click cells to view annotations.
Homozygotes for targeted null mutations exhibit impaired formation of extraembryonic membrane and endoderm and die prior to gastrulation. Heterozygotes develop polyposis of the glandular stomach and duodenum.
Gene Ontology
(GO)
Classifications
less
• All GO Annotations
• GO References
Molecular Function
carbohydrate derivative binding
cytoskeletal protein binding
DNA binding
enzyme regulator
hydrolase
ligase
lipid binding
oxidoreductase
receptor
receptor binding
RNA binding
transcription
transferase
transporter
Biological Process
nucleic acid-templated transcription
carbohydrate derivative metabolism
cell death
cell differentiation
cell proliferation
cellular component organization
establishment of localization
homeostatic process
immune system process
lipid metabolic process
protein metabolic process
response to stimulus
signaling
system development
Cellular Component
cell projection
cytoplasmic vesicle
cytoskeleton
cytosol
endoplasmic reticulum
endosome
extracellular region
Golgi apparatus
mitochondrion
non-membrane-bounded organelle
nucleus
organelle envelope
organelle lumen
plasma membrane
synapse
vacuole
Click cells to view annotations.
Expression
less
Expression Overview
early conceptus
embryo ectoderm
embryo endoderm
embryo mesoderm
embryo mesenchyme
extraembryonic component
alimentary system
auditory system
branchial arches
cardiovascular system
connective tissue
endocrine system
exocrine system
hemolymphoid system
integumental system
limbs
liver and biliary system
musculoskeletal system
nervous system
olfactory system
reproductive system
respiratory system
urinary system
visual system
Click cells to view annotations.
• Assay Results
• Tissues
• cDNA Data
• Literature Summary
Interactions
less
Sequences &
Gene Models
less
Representative SequencesLengthStrain/SpeciesFlank
genomic OTTMUSG00000021090 VEGA Gene Model | MGI Sequence Detail 64772 C57BL/6J ± kb
transcript OTTMUST00000049898 VEGA | MGI Sequence Detail 2233 Not Applicable
polypeptide OTTMUSP00000023202 VEGA | MGI Sequence Detail 551 Not Applicable
For the selected sequence
Polymorphisms
less
• SNPs within 2kb
358 from dbSNP Build 142
• RFLP
Protein
Information
less
Molecular
Reagents
less
• All nucleic 31
Genomic 5
cDNA 19
Primer pair 6
Other 1
Microarray probesets 6
Other
Accession IDs
less
MGD-MRK-33955, MGI:106263, MGI:2147379
References
more
• Summaries
All 316
Developmental Gene Expression 70
Diseases 4
Gene Ontology 49
Phenotypes 180
• Earliest
J:40660 Anna CH, et al., Sequence and chromosomal mapping of the mouse homolog (Madh4) of the human DPC4/MADH4 gene. Mamm Genome. 1997 Jun;8(6):443-4
• Latest
J:240719 Li Y, et al., SMAD3 Regulates Follicle-stimulating Hormone Synthesis by Pituitary Gonadotrope Cells in Vivo. J Biol Chem. 2017 Feb 10;292(6):2301-2314
Contributing Projects:
Mouse Genome Database (MGD), Gene Expression Database (GXD), Mouse Tumor Biology (MTB), Gene Ontology (GO), MouseCyc
Citing These Resources
Funding Information
Warranty Disclaimer & Copyright Notice
Send questions and comments to User Support.
last database update
05/23/2017
MGI 6.09
The Jackson Laboratory | {
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Biotech Science
Possible Monogamy Gene Found In People 440
Calopteryx sends in a New Scientist summary of research from Sweden pointing toward the existence of a gene that influences monogamy in men. (The article doesn't mention women, and the study subjects were all men at least 5 years into a heterosexual relationship.) "There has been speculation about the role of the hormone vasopressin in humans ever since we discovered that variations in where receptors for the hormone are expressed makes prairie voles strictly monogamous but meadow voles promiscuous; vasopressin is related to the 'cuddle chemical' oxytocin. Now it seems variations in a section of the gene coding for a vasopressin receptor in people help to determine whether men are serial commitment-phobes or devoted husbands."
This discussion has been archived. No new comments can be posted.
Possible Monogamy Gene Found In People
Comments Filter:
• Re:Hhhmm, (Score:4, Insightful)
by EnergyScholar ( 801915 ) on Tuesday September 02, 2008 @03:10PM (#24848027)
Just as a guess, which strategy works better (from a 'survival of the genes' perspective) probably varies in different circumstances. This would explain why neither gene sequence has dominated.
• by DoofusOfDeath ( 636671 ) on Tuesday September 02, 2008 @03:11PM (#24848049)
monogamy in general seems to be a mirage
Are you saying that it doesn't exist, or that it's just rarer than we pretend?
Consider that if ~ 50% of married people are adulterous, then there's a huge fraction (~ 50%) who are monogamous.
• Re:Hhhmm, (Score:5, Insightful)
by Millennium ( 2451 ) on Tuesday September 02, 2008 @03:15PM (#24848127)
Shouldn't evolution sided with either monogamy or polygamy? I mean even if there is only a one percent difference between the successor rates should that have not been reflected by now?
If monogamy or the lack thereof were genetic and there were an evolutionary advantage to either strategy, then you're right: that should have been reflected in the general population.
Since it doesn't seem to be, that would seem to indicate that perhaps there is no evolutionary advantage to either side. With no advantage, there is no pressure for humanity to tend in one direction or the other. That could yield a pattern closer to what we are seeing now.
• by Altus ( 1034 ) on Tuesday September 02, 2008 @03:19PM (#24848219) Homepage
Just because someone has the desire to be non-monogamous does not mean that they cheat on their significant others.
• by lena_10326 ( 1100441 ) on Tuesday September 02, 2008 @03:20PM (#24848257) Homepage
And like winning the lottery twice, the slashdot men that do marry are quite unlikely to find another. A predisposition for involuntary celibacy is a predictor for monogomy.
• by t0rkm3 ( 666910 ) on Tuesday September 02, 2008 @03:31PM (#24848441)
Hrmmmm...
As far as I can tell, from literature in polygamist cultures, the jealousy gene is 100% present in females and males of the species. Therefore, it would seem that a barter system would evolve. The higher your wealth is above the mean of the society that you live in, the more likely that you will be able to entice potential partners into a 'mutually beneficial' relationship. The wealth assuages the greater portion of the jealousy, while other services alleviate the remainder.
• by tinkerghost ( 944862 ) on Tuesday September 02, 2008 @03:40PM (#24848585) Homepage
One researcher found that the overwhelming contribution to the increased rate of divorce is the modern concept of marriage for love instead of position/wealth. The current divorce trend is simply the end result of a curve started in the years following the civil war.
So if these conservatives want to go back to an idyllic time with low divorce & happy families - I say bring back arranged marriages.
• Re:Disablites Act (Score:4, Insightful)
by The End Of Days ( 1243248 ) on Tuesday September 02, 2008 @03:45PM (#24848665)
The day nerds become a protected subspecies is the day I give up on humanity.
• by blueg3 ( 192743 ) on Tuesday September 02, 2008 @03:47PM (#24848693)
Why would a man not screw around as much as possible?
In short, because our young are vulnerable after birth, require a fairly large energy investment, and are few in number.
Monogamy actually appears in a number of different animal species.
• Re:Hhhmm, (Score:4, Insightful)
by Hatta ( 162192 ) on Tuesday September 02, 2008 @03:52PM (#24848779) Journal
And the women, they're looking for a powerful man to knock them up, and a nice dedicated man to stick with her and raise a family.
• by AP31R0N ( 723649 ) on Tuesday September 02, 2008 @03:53PM (#24848797)
If humans were meant to be monogamous, we wouldn't be having this conversation. It would be a given and a non-issue. Non-monogamy would be something lesser beasts do and would strike us as odd and curious behavior.
Asking humans to be monogamous is like asking a cat to NOT chase a mouse. "Did you SEE her tits? Of COURSE I hit that. I'd be gay if it didn't!"
Marriage is a system invented by men with power to make alliances and to manage inheritance of power. The whole love thing is very 20th century.
• by vux984 ( 928602 ) on Tuesday September 02, 2008 @03:55PM (#24848827)
Let's say that we go 10,000 years back. Why would a man not screw around as much as possible?
Lots of reasons...
Inability to find good mates... Ideal mothers for your children would reject you knowing that you wouldn't provide for them?
Low chance of offspring surviving... mothers would be unable to care for your children, and unable to find mates willing help them?
Societal acceptance... e.g. The other men would stone him? Stone the women he cheated with? Stone his offspring?
Monogamy exists in nature. There are reasons for why it works where it exists.
And if love existed, who's to say that it lasted for long periods?
Indeed. Monogamy isn't necessarily 'till death to we part' in modern society at least it simply means not cheating on your partner. It is entirely possible to marry, raise a child, separate, marry someone else, and even raise another child, all within the confines of monogamy.
Hell when I was a teen, most of us were pretty monogamous; its not that we all married our first crush, but rather that our teen years were a succession of monogamous relationships of varying lengths, some quite brief, and punctuated with periods of being 'single'.
And yes some people who were supposedly 'in a relationship' cheated, and when caught it carried a stigma, one that I would say definitely impacted their dating prospects in the circles where it was known that they cheated (applied to both males and females).
• by Anachragnome ( 1008495 ) on Tuesday September 02, 2008 @04:01PM (#24848921)
Hate to kick the barstool out from under y'all, but Jeebuz, you folks act like a gene sequence removes all thought from the equation.
I don't sleep around because I love my wife and extra-marital affairs have a tendency to remove MARRIAGES. Quite frankly, it is my head, and the thoughts within, that decide my actions, not the genes passed on to me. Genes may have some effect, but if the result is not acceptable to the thinking part of me, they are simply over-ridden.
• by KGIII ( 973947 ) <uninvolved@outlook.com> on Tuesday September 02, 2008 @04:03PM (#24848959) Journal
They lie to you. Oh man do they lie. They probably do it so that you will join them in their misery, misery loves company and all that.
Anyhow, now that there's a gene for it and I obviously don't have it, I have a scientific excuse. ;)
• Re:Hhhmm, (Score:5, Insightful)
by megaditto ( 982598 ) on Tuesday September 02, 2008 @04:03PM (#24848977)
I think you might be wrong there.
In a welfare/socialist society, polygamy and promiscuety make more (evolutionary) sense for men.
Which would you rather be: 1) the guy that sleeps around with lots of women and gets lots of kids, or 2) the guy that stays with a single woman and gets taxed to death to support all the single mothers, left over from the first guy.?
• Re:Hhhmm, (Score:2, Insightful)
by Anonymous Coward on Tuesday September 02, 2008 @04:23PM (#24849305)
Genetics is becoming the new astrology. Seriously. It's a good analogy too, sure stellar bodies affect our natural worlds, and sure genetics affects our bodies and minds, but I have not seen any proof that theses genetic changes are significant to human behavior on a day to day level.
Sure an animal living on instinct, slave to the chemicals sloshing around in it's brain pan, has it's mating style dictated by genes, but humans are just a little smarter then that.
I used to be a Beta male and used to pursue monogamy through out high school and early college for exactly the same reason you describe. Then I started working construction, got buff and learned how to quote "play the field" and have done things that belong on Tucker Max, not /., but I changed and I didn't need gene therapy to do it, why? because I think, and I can rationalize and weigh consequences against rewards, so while I may feel guilty for sleeping around, I know it's way funner than having to put up with some cow.
Humans are led by their frontal lobe and will ignore all those little subconsciouses chemicals in favor of what ever is easiest. I'm sure this gene might have an affect, but I'm sure on full blast the best difference it would make would be to cause a guy to buy his wife flowers after he cheats on her. Smoking a cig has a stronger kick.
The reason the gene is still around and hasn't gone one way or the other is because it's just genetic kruft that doesn't matter.
• by OeLeWaPpErKe ( 412765 ) on Tuesday September 02, 2008 @04:23PM (#24849319) Homepage
Or rather : in different circumstances monogamy may provide an evolutionary advantage. If you intend to wage unceasing war ("live off the land" (and the passersby)), for example, monogamy would be a bad idea, since lots of men will die, leaving behind women, even though some limit would be good (say you expect 50% of the men to be involved in war, then you should allow 2 women to one man, if you expect between 75 and 90% of your society to be dedicated to war, then 4 women to 1 man seems appropriate (and obviously only to men who can afford not to be on the frontlines, who should basically stay away from women, except the occasional rape of a succesful raid) (then again, in war, are limits like these really going to be respected ?). If you allow without limit (or allow polygamy + concubines) then clearly you expect to do nothing else than warfare, and marriage means nothing except for inheritance.
In peace, you'd need to prevent men remaining behind alone without partner (because for every extra woman one man has, another has to do without, 4 women to one man would become 75% of men without contact with women in extremis, realistically, say 50% of men, 4 women + unlimited (and exclusive) concubines would mean something like 999/1000 of men without partner, in some cultures that is normal, or was normal not too long ago), as that will certainly not be helpful in helping them build instead of destroying society, therefore in a peaceful setting, you'd want monogamy.
The fact that genes start expressing it is not very surprising. Polygamous cultures are known for being more than a little agressive, and genes are how humans adapt to their environment. If the environment or the culture changes to be less suitable for agriculture (or the culture doesn't know, or incorrectly conducts agriculture, e.g. predatory agriculture, or not doing anything about overpopulation, or ...) the genes will adapt to become less monogamous.
If raiding is basically impossible, for whatever reason, building things will become important, and monogamous relationships become an evolutionary advantage. Certainly after 10 generations the effects will be very noticeable.
Since this gene will very much influence how agressive people are against "other tribes", it is one of the prime parameters that will determine the layout of the resulting society, and may introduce all sorts of limits (e.g. agressive societies will never have any population density for obvious reasons, which can easily translate in a very low maximum population limit)
• Re:Hhhmm, (Score:3, Insightful)
by Mr. Slippery ( 47854 ) <{tms} {at} {infamous.net}> on Tuesday September 02, 2008 @04:42PM (#24849721) Homepage
Which would you rather be: 1) the guy that sleeps around with lots of women and gets lots of kids, or 2) the guy that stays with a single woman and gets taxed to death to support all the single mothers, left over from the first guy.?
How about 3) the guy that sleeps around with lots of women and has no kids?
Effective birth control exists. Use it.
• by Mr. Slippery ( 47854 ) <{tms} {at} {infamous.net}> on Tuesday September 02, 2008 @04:47PM (#24849835) Homepage
Genes may have some effect, but if the result is not acceptable to the thinking part of me, they are simply over-ridden.
The point is that the genes determine, or at least influence, what the "thinking part" of you find acceptable.
You are not consciousness inhabiting a body. You are consciousness generated by a body. The nature of that consciousness is determined by the nature of the body. The nature of the body is determined by genetics and by environment.
• by Anonymous Coward on Tuesday September 02, 2008 @05:57PM (#24851055)
The poster didn't say anything about marriage. He just said "monogamous relationships."
"Living together in sin" qualifies as a monogamous relationship, and it very well may include frequent sex and home-cooked meals. In fact, remaining unmarried in such a relationship could be quite fulfilling for both partners while avoiding the marriage tax penalty and other legal complications.
Furthermore, each partner will feel free to leave should things turn sour (rather than trapped by a contract and economic devastation from an expensive divorce). This could cultivate a much healthier state of mind (while avoiding the "I don't have to work so hard to please you now, because we're married!" mentality) which will keep the couple together even longer than if they got married.
I am curious to know, however, why you think marriage is misery. Worse than a car accident? In what way? Is it because your spouse pulled a bait-and-switch on you, becoming someone entirely different once the contract was signed? Sounds to me like you just picked the wrong person....
• by Crazy Taco ( 1083423 ) on Tuesday September 02, 2008 @06:40PM (#24851675)
They lie to you. Oh man do they lie. They probably do it so that you will join them in their misery, misery loves company and all that.
Actually it's not a lie. Like everything, there are of course exceptions, so I'm sure some people do lie, but I get all of the above. And my wife is wonderful. I love being married. So yeah, if you wanna delude yourself out of the fun, go ahead... but consider the fact that you haven't tried being married, so how would you know the truth? Married men are inherently credible when talking about this issue, and unmarried men are without credibility, for the following reason: all us married men have been both single and married, and I personally can say that after trying both, marriage is far superior.
Anyhow, now that there's a gene for it and I obviously don't have it, I have a scientific excuse. ;)
Sorry, but as a creature of reason and logic (which your appeal to science shows that you are), you are still without excuse. Anyone who claims reason has the tools necessary to rise above base animal instincts and live differently. Whether this alleged gene actually is proven to be true or not, the "serial non-commiters" still have no excuse to use the women around them.
As humans, we have to rise above this non-commitment, because regardless of a specific gene, societies in which commitments are made and upheld are inherently more stable and peaceful than those in which no one can trust anyone's word. As humans, our goal should be to form stable societies that are best for us, not to follow our genetic dispositions.
• Re:Disablites Act (Score:3, Insightful)
by budgenator ( 254554 ) on Tuesday September 02, 2008 @08:30PM (#24852945) Journal
Oh yeah nerds in PR are going to hurt the company a lot more than the narcissistic PHBs in management will.
• by zobier ( 585066 ) <zobier@@@zobier...net> on Wednesday September 03, 2008 @03:25AM (#24855847)
As humans, we have to rise above this non-commitment, because regardless of a specific gene, societies in which commitments are made and upheld are inherently more stable and peaceful than those in which no one can trust anyone's word. As humans, our goal should be to form stable societies that are best for us, not to follow our genetic dispositions.
Except that we're talking about monogamy, not commitment. It is possible to be in a committed relationship, have sexual relations with more than one person and not be dishonest about it. Since we're talking about logic, reason and whatnot. It is also quite possible to have a stable and trusting society where this behaviour is not taboo.
"The way of the world is to praise dead saints and prosecute live ones." -- Nathaniel Howe
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6,498,267,451,030,723,000 | 2019 O'Hara Fellowship Awarded to Bioinformatics Student
Bioinformatics doctoral student Alli Gombolay was selected for the 2019 O'Hara Fellowship. This fellowship is presented to top doctoral students in the College of Sciences following a nomination from each of the Schools in the College. The O'Hara Fellowship includes a financial award for each semester in 2019. Alli's research focuses on Ribose-Map, a bioinformatics toolkit to profile rNMPs embedded in DNA. Her research advisor is Professor Francesca Storici. | {
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-459,052,904,039,989,570 | Written by John Tyler Bonner
reproduction
Article Free Pass
Written by John Tyler Bonner
reproduction, process by which organisms replicate themselves.
In a general sense reproduction is one of the most important concepts in biology: it means making a copy, a likeness, and thereby providing for the continued existence of species. Although reproduction is often considered solely in terms of the production of offspring in animals and plants, the more general meaning has far greater significance to living organisms. To appreciate this fact, the origin of life and the evolution of organisms must be considered. One of the first characteristics of life that emerged in primeval times must have been the ability of some primitive chemical system to make copies of itself.
At its lowest level, therefore, reproduction is chemical replication. As evolution progressed, cells of successively higher levels of complexity must have arisen, and it was absolutely essential that they had the ability to make likenesses of themselves. In unicellular organisms, the ability of one cell to reproduce itself means the reproduction of a new individual; in multicellular organisms, however, it means growth and regeneration. Multicellular organisms also reproduce in the strict sense of the term—that is, they make copies of themselves in the form of offspring—but they do so in a variety of ways, many involving complex organs and elaborate hormonal mechanisms.
Levels of reproduction
Molecular replication
The characteristics that an organism inherits are largely stored in cells as genetic information in very long molecules of deoxyribonucleic acid (DNA). In 1953 it was established that DNA molecules consist of two complementary strands, each of which can make copies of the other. The strands are like two sides of a ladder that has been twisted along its length in the shape of a double helix (spring). The rungs, which join the two sides of the ladder, are made up of two terminal bases. There are four bases in DNA: thymine, cytosine, adenine, and guanine. In the middle of each rung a base from one strand of DNA is linked by a hydrogen bond to a base of the other strand. But they can pair only in certain ways: adenine always pairs with thymine, and guanine with cytosine. This is why one strand of DNA is considered complementary to the other.
The double helices duplicate themselves by separating at one place between the two strands and becoming progressively unattached. As one strand separates from the other, each acquires new complementary bases until eventually each strand becomes a new double helix with a new complementary strand to replace the original one. Because adenine always falls in place opposite thymine and guanine opposite cytosine, the process is called a template replication—one strand serves as the mold for the other. It should be added that the steps involving the duplication of DNA do not occur spontaneously; they require catalysts in the form of enzymes that promote the replication process.
Molecular reproduction
The sequence of bases in a DNA molecule serves as a code by which genetic information is stored. Using this code, the DNA synthesizes one strand of ribonucleic acid (RNA), a substance that is so similar structurally to DNA that it is also formed by template replication of DNA. RNA serves as a messenger for carrying the genetic code to those places in the cell where proteins are manufactured. The way in which the messenger RNA is translated into specific proteins is a remarkable and complex process. (For more detailed information concerning DNA, RNA, and the genetic code, see the articles nucleic acid and heredity: Chromosomes and genes). The ability to synthesize enzymes and other proteins enables the organism to make any substance that existed in a previous generation. Proteins are reproduced directly; however, such other substances as carbohydrates, fats, and other organic molecules found in cells are produced by a series of enzyme-controlled chemical reactions, each enzyme being derived originally from DNA through messenger RNA. It is because all of the organic constituents made by organisms are derived ultimately from DNA that molecules in organisms are reproduced exactly by each successive generation.
Cell reproduction
The chemical constituents of cytoplasm (that part of the cell outside the nucleus) are not resynthesized from DNA every time a cell divides. This is because each of the two daughter cells formed during cell division usually inherits about half of the cellular material from the mother cell (see cell: Cell division and growth), and is important because the presence of essential enzymes enables DNA to replicate even before it has made the enzymes necessary to do so.
Cells of higher organisms contain complex structures, and each time a cell divides the structures must be duplicated. The method of duplication varies for each structure, and in some cases the mechanism is still uncertain. One striking and important phenomenon is the formation of a new membrane. Cell membranes, although they are very thin and appear to have a simple form and structure, contain many enzymes and are sites of great metabolic activity. This applies not only to the membrane that surrounds the cell but to all the membranes within the cell. New membranes, which seem to form rapidly, are indistinguishable from old ones.
Thus, the formation of a new cell involves the further synthesis of many constituents that were present in the parent cell. This means that all of the information and materials necessary for a cell to reproduce itself must be supplied by the cellular constituents and the DNA inherited from the parent cell.
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8,713,176,549,789,221,000 | Document Detail
Mycosporine glycine protects biological systems against photodynamic damage by quenching singlet oxygen with a high efficiency.
MedLine Citation:
PMID: 12945577 Owner: NLM Status: MEDLINE
Abstract/OtherAbstract:
This report concerns physiological function of mycosporine-like amino acids (MAA) as an active defense against the photooxidative effects of sunlight in marine organisms. Mycosporine glycine (MG) is a representative member of MAA family and was found to effectively suppress various detrimental effects of the Type-II photosensitization in biological systems, such as inactivation of mitochondrial electron transport, lipid peroxidation of microsomes, hemolysis of erythrocytes and growth inhibition of Escherichia coli. The presence of MG in solutions of eosin Y or methylene blue resulted in a marked decrease in the level of singlet oxygen (1O2) produced by the sensitizers under illumination. The rate constant of 1O2 quenching by MG was determined to be 5.6 x 10(7) M(-1) s(-1) by the time-resolved 1O2 luminescence decay method, which is higher than, or at least comparable to, the values for 1O2 reaction of well-known quenchers such as 1,4-diazabicyclo[2,2,2]octane and furfuryl alcohol. The results suggest that MG probably together with some other active MAA may play an important role in protecting marine organisms against sunlight damage by eliminating 1O2 generated from certain endogenous photosensitizers.
Authors:
Hwa-Jin Suh; Hyun-Woo Lee; Jin Jung
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Publication Detail:
Type: Journal Article; Research Support, Non-U.S. Gov't
Journal Detail:
Title: Photochemistry and photobiology Volume: 78 ISSN: 0031-8655 ISO Abbreviation: Photochem. Photobiol. Publication Date: 2003 Aug
Date Detail:
Created Date: 2003-08-29 Completed Date: 2003-12-02 Revised Date: 2006-11-15
Medline Journal Info:
Nlm Unique ID: 0376425 Medline TA: Photochem Photobiol Country: United States
Other Details:
Languages: eng Pagination: 109-13 Citation Subset: IM
Affiliation:
School of Agricultural Biotechnology, Seoul National University, Suwon, South Korea.
Export Citation:
APA/MLA Format Download EndNote Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Aplysia / radiation effects
Escherichia coli / growth & development, radiation effects*
Glycine / pharmacology*
Hemolysis / radiation effects
Humans
Kinetics
Phytoplankton / radiation effects
Sea Cucumbers / radiation effects
Seawater
Singlet Oxygen*
Submitochondrial Particles / radiation effects
Sunscreening Agents / pharmacology*
Urochordata / radiation effects
Chemical
Reg. No./Substance:
0/Sunscreening Agents; 17778-80-2/Singlet Oxygen; 56-40-6/Glycine
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Intrinsic fluorescence polarization of amniotic fluid: evaluation of human fetal lung maturity.
Next Document: Inter- and intraspecific variation in excited-state triplet energy transfer rates in reaction center... | {
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-9,127,614,458,070,122,000 | lunes, 5 de febrero de 2018
Researchers shine light on ‘molecular butcher’ using neutron beams
Researchers shine light on ‘molecular butcher’ using neutron beams
News-Medical
Researchers shine light on ‘molecular butcher’ using neutron beams
Tucked away inside cell membranes, a molecular butcher does the bidding of healthy cells but also of disease agents. It has been operating out of clear view, but researchers just shined a mighty spotlight on it.
The butcher is a common enzyme called presenilin, which chops lengthy protein building blocks down to useable shorter lengths. It resides in membrane spaces that evade ready experimental detection, but in a new study, researchers at the Georgia Institute of Technology and Oak Ridge National Laboratory (ORNL) have illuminated presenilin using a neutron beam produced by the world's most powerful research nuclear reactor.
"One third of our genome goes to work to encode intramembrane proteins," said Raquel Lieberman, an associate professor in Georgia Tech's School of Chemistry and Biochemistry. "Some of them are huge and do super complex biochemistry."
The enzyme presenilin in particular is an intramembrane protease. There are four classes of these, and they are needed, among other things, for: Alerting to and defending against infectors, and cell differentiation and development.
If the latter two go wrong, that can lead to cancer.
Grainy neutron mugshot
Now, the researchers have gotten a figurative mugshot of one intramembrane protein, the presenilin. Technically speaking, the researchers worked with a presenilin cousin found in microbes -- M. marisnigri intramembrane aspartyl protease or MmIAP -- but here we will use presenilin and MmIAP interchangeably for simplicity's sake.
The measurement was low-resolution but revealed enough to establish that the protein structure is more simply put together than previously believed, and that surprised the scientists.
"Our sample shows that this is a monomer all by itself," Lieberman said. "We were expecting a dimer or a trimer." That means it was made up of one long strand, mostly coiled up like a spring, instead of doubled-up or tripled-up curly strands.
Presenilin (MmIAP) is armed with two chemical knives, aspartates, that reliably make cuts on peptides, the subunits that make up proteins. And a second new study by the same researchers illuminated how the cleaving works.
Anybody's peptide butcher
Presenilin can trim peptides into building blocks helpful to its own cells, or whittle bad peptide chunks that end up in amyloid-beta plaque, a suspect in Alzheimer's disease. Or presenilin can aid and abate hepatitis C viruses by carving components it needs to reproduce.
Understanding how presenilin works could one day prove useful to medical research. "If you could find a way to interfere with it selectively, you could stop the spread of hepatitis C in the body," Lieberman said.
The researchers, led by Lieberman and neutron scattering scientist Volker Urban from ORNL, published the revelations of the neutron scattering on February 2, 2018, in Biophysical Journal. The new insights into presenilin functioning is to officially publish in March in the Journal of Biological Chemistry, but is currently available online without embargo. First authors were Swe-Htet Naing of Georgia Tech and Ryan Oliver of Oak Ridge.
Research was funded by the National Science Foundation, the National Institutes of Health, and the U.S. Department of Energy.
Herding hydrophobic hiders
The scientists reached for the big gun when they went to the High Flux Isotope Reactor (HFIR) to make presenilin (MmIAP) come out of hiding.
HFIR's neutron beams were cooled to minus 253 degrees Celsius (minus 424 degrees Fahrenheit) to slow the neutrons down, so they could probe molecular features of the biological samples.
Presenilin and other intramembrane proteins warrant such proverbial desperate measures. They live in a lipid environment and hate water about the way cats do, and that's a problem for researchers studying them.
"When you have proteins that are not soluble in water, you're in trouble," Lieberman said. "The usual techniques to analyze them become very, very difficult, if not impossible. And when you chemically bootstrap these proteins to be able use these water-soluble methods, you have really poor chances of seeing the protein's actual structure that performs its function."
Form follows function
Images derived from water-based analytical methods in Lieberman's lab have not completely jibed with presenilin's function. For one, the enzyme's cutting surfaces have been too far apart. The neutron beam's revelations made more sense to the researchers.
"Our shape was tighter, and made more sense with presenilin's function in its natural setting in the membrane," Lieberman said.
The presenilin (MmIAP) samples examined at the HFIR were suspended in a solution friendly to the hydrophobic protein. Ironically, presenilin and other intramembrane proteases often hydrolyze peptides, in other words, they add water to them.
"These proteases are confined to the lipid cell membrane where there is no water. Since water is required for hydrolysis, it has to come from outside the membrane," Lieberman said. "How that happens is yet another mystery that needs uncovering."
Robust, reliable choppers
The precision and consistency, with which the presenilin homologue MmIAP cleaved peptides, impressed the researchers.
"When we used a model synthetic peptide, it cleaved only at very specific positions on the peptide," Lieberman said. "When we switched to a real biological peptide, it also cleaved very exactly."
The researchers put the presenilin through various mutations, which had little to no effect on its cleaving abilities. That could mean that its baseline functioning is nearly immune to genetic interference.
On a chilling note, when cutting amyloid-beta precursor peptides, the researchers observed the microbial presenilin cousin, MmIAP, always making the chop in a way notorious for amyloid's association with Alzheimer's disease.
"We never saw the cut that made what is typically viewed as the 'good' amyloid, A-beta-40," Lieberman said. "We only saw cuts that led to the 'bad' amyloid, A-beta-42."
More research would be needed to explain why that happened; if the same is true for presenilin in human cell membranes, and also if some regulator prevents the creation or accumulation of so much bad amyloid in healthy cells.
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6,618,616,839,048,270,000 | World Environment Day: Over £8m boost for international conservation
0
370
Over £8 million new funding to protect rare wildlife and vulnerable habitats across the globe.
Threatened species such as whales, marine turtles and sharks will be better protected thanks to a boost of over £8 million for projects in the UK Overseas Territories, the government has announced today (Saturday 5 June) under plans to tackle the global biodiversity crisis.
The funding will also help protect a number of rare species and vulnerable habitats across the globe from the threats from invasive species.
Over the next three years, 31 projects will receive £8.02 million through the Darwin Plus scheme for conservation of the unique and globally significant environments found in UK Overseas Territories.
Habitats and species set to benefit from funding include:
• Threatened albatross species in the southern Atlantic overseas territories, Tristan da Cunha, South Georgia and the South Sandwich Islands, through improved population monitoring
• A number of species found in Cayman’s Sister Islands will be safeguarded from invasive species such as feral cats and invasive green iguanas
• Anguilla’s shark populations, through increasing knowledge of their habitats and conservation needs, while developing local ownership of their conservation through greater community engagement
• The Ascension Islands’ marine turtles through making improvements to their monitoring programme using innovative modelling techniques and new labour-saving technologies
• Coral reefs in the Indian Ocean by helping small-scale fishers to sustainably manage these habitats on the island of Diego Garcia
International Environment Minister Lord Goldsmith said:
World Environment Day provides us all with a stark reminder of why we need to take urgent action to reverse global biodiversity loss.
The Darwin Plus funding announced today will support the magnificent biodiversity hotspots that make up our Overseas Territories, which are so threatened by climate change. It will restore precious ecosystems, prevent the extinction of some of the world’s most wonderful species, and at the same time transform the lives of the poorest communities.
Over the last decade the Darwin Plus programme has supported over 120 individual projects supporting conservation in marine, terrestrial and freshwater environments. The UK government, as president of the G7 and COP26, is leading the way globally in the fight to tackle climate change and repair the natural world.
Professor E.J. Milner Gulland, Oxford University and Chair of the Darwin Expert Committee and Darwin Plus Advisory Group, said:
The UK’s Overseas Territories are home to some of our most iconic and important threatened biodiversity, as well as rich and productive natural resources. The Darwin Plus projects span the range of biodiversity from wetlands to whales, and addresses issues from controlling invasive thorns to tracking threatened turtles. So I’m really happy that, on World Environment Day, the Darwin Plus fund is supporting the Overseas Territories to conserve their precious biodiversity while also building a sustainable future for people and nature.
Beccy Speight, Chief Executive of the RSPB said:
Our Government has an important role to play as we all work to revive our world, the UK’s Overseas Territories are home to 94% of the plants and animals that are only found on UK soil. But these amazing places are under threat from the nature and climate emergencies.
Failing to act in our Territories would raise the real risk of global extinctions, so this vital funding will help fulfil our responsibilities to protect our precious wildlife, from tropical rainforests in the Caribbean to wind-swept albatross islands in the Southern Ocean. Today’s announcement, plus the UK Government’s additional £1.5m contribution to support our major partnership project to restore Gough Island, a threatened UK World Heritage Site in the South Atlantic recognised as one of the most important seabird islands in the world, will be welcome news to the local community and many individuals who continue to support this vital work.
Last month, the Climate and Environment Ministers of the G7 committed to halting and reversing the loss of biodiversity by 2030. In efforts to tackle the twin challenges of climate change and biodiversity loss, all G7 members also signed up to the global ‘30×30’ initiative to conserve or protect at least 30 per cent of the world’s land and at least 30 per cent of the world’s ocean by 2030, as well as committing to ‘30×30’ nationally.
The funding being announced today builds on the £220 million for biodiversity conservation in developing nations, and the doubling of UK international climate finance, announced by the Prime Minister at the UN General Assembly in 2019.
Today’s announcement forms part of the Government’s commitments to drive international ambition on action to tackle the biodiversity crisis and work towards nature-based solutions ahead of the G7, the upcoming 15th UN Biodiversity Conference of the Parties (Convention of Biological Diversity COP15), and the 26th UN Climate Change Conference of the Parties (COP26) which will be hosted in Glasgow later this year. In the run up to the summit, the UK is focused on four goals to drive progress: securing global net zero, protecting communities and natural habitats from the impacts of climate change, mobilising finance and working together to accelerate action.
A full list of projects, including a number of small schemes, to be supported by the Darwin Plus programme is available on the Darwin Initiative website.
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938,048,742,893,902,300 | Renhao Li Lab:Gallery
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(Group photos)
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[[Image:RenhaoLiLab_BBQ2013_1.jpg|600px]]
[[Image:RenhaoLiLab_BBQ2013_1.jpg|600px]]
-
Good luck at Vanderbilt, Alex!
+
We missed you, Sungyun.
Revision as of 11:09, 16 August 2013
Home Research Publications Protocols Lab Members Gallery
Contents
Research Images
Xi's paper on the interaction between GPV and GPIb-IX made the cover of the September, 2012 issue of Journal of Thrombosis and Haemostasis.
Rong's paper showing the reconstitution of GPIb-IX complex into nanodiscs was featured on the Biochemistry journal web site.
Wenjun's paper on the organization of GPIb-IX complex was highlighted by a commentary in Blood entitled "Structural mix-n-match reveals molecular secrets of platelets". The above image was in the commentary.
Group photos
BBQ, 2013
We missed you, Sungyun.
Where is BBQ?
Bowling, 2012
You know we could've photoshopped it.
Going at it
Oops ... too close
The Move, 2011
Boxes Arrived
Boxes in the hallway
Lab into boxes
Office into boxes
Freezers loaded into the truck
Lab boxes loaded into the truck
Office boxes loaded into the truck
Going, going, gone!
Personal tools | {
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4,358,969,078,787,250,700 | brinklifescience Ms. Hall
Brink Junior High Life Science Teacher 7th Grade
http://www.telepath.com/mps/brink
WELCOME TO LIFE SCIENCE WEB PAGE FOR S. HALL
ANNOUNCEMENTS:
There will be a TEST Wed. April 5, 2000.
SUBJECT: Chapter 16 only over the eye and ear and all the vocabulary words. The voc will be a matching test. The eye and ear will be a labeling test.
Last updated 2008/09/28 11:12:13 PDTHits 191 | {
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-600,148,046,248,958,600 | The International Conference on Intelligent Biology and Medicine (ICIBM) 2016: Putting systems biology to work
W. Jim Zheng, Jianhua Ruan, Hua Xu, Zhongming Zhao, Zhangdong Liu
Research output: Contribution to journalEditorialpeer-review
Abstract
Between December 8-10, 2016, the International Conference on Intelligent Biology and Medicine (ICIBM 2016) was held in Houston, Texas, USA. The conference included eight scientific sessions, four tutorials, one poster session, four highlighted talks and four keynotes that covered topics in 3D genome structure analysis and visualization, next generation sequencing analysis, computational drug discovery, medical informatics, cancer genomics and systems biology. Systems biology has been a main theme in ICIBM 2016, with exciting advances were presented in many areas of systems biology. Here, we selected seven high quality papers to be published in BMC Systems Biology.
Original languageEnglish (US)
Article number88
JournalBMC Systems Biology
Volume11
DOIs
StatePublished - Oct 3 2017
ASJC Scopus subject areas
• Structural Biology
• Modeling and Simulation
• Molecular Biology
• Computer Science Applications
• Applied Mathematics
Fingerprint
Dive into the research topics of 'The International Conference on Intelligent Biology and Medicine (ICIBM) 2016: Putting systems biology to work'. Together they form a unique fingerprint.
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-3,294,706,627,876,457,000 | Jonathan Lewis
Multinational tropical microbiology
Medellín has an unfortunate historical reputation for violence, but it is now emerging as a vibrant and modern Andean destination for travellers, so it sounded like the perfect location to carry out a two week microbiological research project. As such a group of British and American students and professors joined up with counterparts from the University of Antiquia and EAFIT University in Medellín to broaden the horizons of everyone involved on both a scientific and cultural level.
Our first encounter with the American students saw us sharing a lively evening at a Colombian restaurant, with most of us eating various local dishes, and a handful eating very un-traditional ribs and chips, an introduction second to none. This proved to be an excellent icebreaker and allowed us to form very close bonds with them, forging many friendships, some, I’m sure, may prove to be lifelong. The very next day we corralled ourselves to meet with the Colombian students and found that upon conversing with them we found we shared many interests, and, as such broken Spanglish became more commonly heard from both British, American, and Colombian mouths.
During our short stay in the country we were split into small, multinational groups, before being let loose to sample as many soil bacteria with antifungal properties from the local populace as possible. Each group harvested five samples from two different locations around Medellín one of which is shown in the group picture of everyone who went below. Another set of five were taken from a rainforest further afield – although all three locations were nature reserves. While in the field we learnt a lot about the effect of the soil on the local environment. After which, we deemed our acquisition complete, so retired to a laboratory within the Universidad de Eaifit to perform our experiments.
groupPutting all personal feelings for the bacteria aside, we got to work; through broken plates, negative results, and mountains of agar we toiled. However – eventually we were able to procure enough valid data to provide the actual researchers an inroad to the antifungal properties of the bacteria lucky enough to be plated out. As a consequence of the multinational groups in which we were placed; we were lucky enough to learn different approaches to techniques we were already somewhat familiar with, such as plate streaking. Within the laboratory itself we were working alongside undergraduate, masters and PhD students and as such were able to learn from each other as well as from the professors who accompanied us.
The trip was an absolute blast, however for me, what topped it off was during the birthday celebration of a Colombian professor, a large bug wondered into the room and perched on a door frame. Everyone in the room gravitated towards it, from the celebrations, and at that point I finally comprehended that despite heralding from different parts of the world, we were all united in our love of nature.
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8,390,941,718,473,794,000 | MTFMT Antibody (1A10)
Images
Product Details
Summary
Product Discontinued
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Order Details
• Catalog Number
NBP2-01231
• Availability
Product Discontinued
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MTFMT Antibody (1A10) Summary
Immunogen
Human recombinant protein fragment corresponding to amino acids 175-389 of human MTFMT(NP_640335) produced in E.coli.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
MTFMT
Purity
Protein A or G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
Packaging, Storage & Formulations
Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Conjugate
Unconjugated
Buffer
PBS (PH 7.3) containing 1% BSA, 50% glycerol
Preservative
0.02% Sodium Azide
Concentration
Please see the vial label for concentration. If unlisted please contact technical services.
Purity
Protein A or G purified
Reactivity Notes
Human
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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Publications for MTFMT Antibody (NBP2-01231) (0)
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Ask a Scientist - We have a lab full of white coats just waiting for your scientific questions and concerns. | {
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1,139,049,899,174,684,700 | Life on Earth
The timeline of life evolving on Earth
Credit: LadyofHats
Life on Earth has taken billions of years to evolve. Life is characterised as anything which has the following traits:
• Move: all living things can move in some way. Plants may look stationary, but they are able to move some parts of their structure.
• Respire: a lot of people think respiration is breathing. Really, it is a chemical reaction which converts glucose, a type of carbohydrate, to energy. All living things respire. To do this most use oxygen, but others use elements like nitrogen.
• Sense: this is when a living thing responds to its environment. If it gets cold we shiver, or if it gets hot we sweat. Seeds will sprout when spring arrives and the soil warms.
• Grow: all living things grow with time. This generally means increasing in mass.
• Reproduce: this is how new living things are created. It can be as simple as one cell splitting into two. Or, it can be more complex, with parts of cells from male and female parents fusing.
• Excrete: this is how waste products are removed from living things, like urine. All living things produce some waste. If this waste stays inside it becomes toxic. All living things have a way of removing waste products.
• Take in nutrients: all living things need to take in food. The energy from food allows them to complete the functions we have mentioned. They need to complete all these functions to survive.
Planet Earth was formed around 4.5 billion years ago. The first signs of life began about 3.6 billion years ago, in the ocean. The water in the ocean protected organisms living there from the dangerous rays of the sun. These x-rays and gamma-rays are part of the electromagnetic spectrum, which damage living things. Later, Earth developed an atmosphere. This protected living things on the land, and life continued to evolve.
Humans, as the species we are now, homo sapiens, have only been around on this planet for around one million years. | {
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-2,121,540,759,459,048,200 | Higher Hyde Heath © James Hitchen
Higher Hyde Heath © James Hitchen
Silver-studded blue © James Hitchen
Silver-studded blue © James Hitchen
Grayling © James Hitchen
Grayling © James Hitchen
An internationally important area of dry & wet lowland heathland & woodland
Location
Bere Regis
Wareham
Dorset
BH20 7NY
OS Map Reference
SY8542889938
A static map of Higher Hyde Heath
Know before you go
Size
56 hectares
Entry fee
N/A
Parking information
Car park via unlocked field gate.
Bicycle parking
Yes
Grazing animals
Pony and cattle grazing all year.
Walking trails
Some paths from the car park. Otherwise Open Access.
Access
Wheelchair access to hide/ viewing area via level track. Access to the rest of the reserve along uneven paths/ pedestrian gates & stiles.
UNDER THE COUNTRYSIDE AND RIGHTS OF WAY ACT 2000 (CROW), DOGS SHOULD BE KEPT ON SHORT LEADS ON THIS SITE (OPEN ACCESS LAND) FROM 1ST MARCH TO 31ST JULY TO PROTECT GROUND NESTING BIRDS.
There is an information board at the car park entrance with a map of the site, and a surfaced path to a hide with viewing area overlooking a pond. This path continues around the pond from the hide and leads out over a stile onto the heath. A path also leads down into the wood before reaching the hide, this also follows out onto the open heath through a pedestrian gate. All unsurfaced paths may be wet and uneven in places. Although the site is Open Access please be aware that sensitive heathland species, including ground nesting birds, may occur throughout. The ground is generally rough and uneven away from the paths, and several ponds and wet areas are present on site.
Ponies are used to graze a large part of the site all year. Please keep your distance, and observe any signs. Adders and ticks are also present. Please see our Visitor Information page for more details.
Dogs
On a lead
Please remove all dog mess from site.
When to visit
Opening times
Open at all times
Best time to visit
Spring and summer
About the reserve
Higher Hyde Heath is a gem for reptile fans. It’s an internationally important heath which is home to all six of the UK’s native reptiles.
Nature fans should tread quietly and may be lucky enough to spot an adder, the only venomous snake in the UK. Don’t be alarmed however, these are very shy and non-aggressive, if undisturbed. They have beautiful and distinctive crisscross markings along their backs. Explorers might also find the very rare smooth snake (we are lucky in Dorset that they love heathland), grass snake (with its yellow and black collar), the common lizard, slow worm (which isn’t actually slow or a worm!) or the extremely rare sand lizard.
All the different types of heathland environment - dry, wet and humid are present, as well as a selection of peaty pools, a mire and wet woodland to explore.
Aside from the wonderful reptile life, there are also lots of rare and interesting heath dwellers at the site, including the ground-nesting nightjar, Dartford warbler, woodlark and tree pipit. Many interesting dragon and damselflies live in the wetter areas, whilst grayling and silver-studded blue butterflies can be found on the open heath. The heath is home to a variety of typical species including common heather, bell heather and gorse throughout, with bog asphodel, sphagnum mosses and pale butterwort in the wetter heath. One to watch out for is the fascinating sundew. These carnivorous plants have sticky sap which trap flies and small insects.
The sundew's tendrils detect the presence of its stuck prey and curl inwards to engulf it. Eventually, the whole leaf wraps around the insect which is digested. The acidic habitats the Round-leaved Sundew lives in on the heath don't provide enough nutrients, so it has evolved this carnivorous way of life to supplement its diet.
Useful Information
4km north of Wool, access from Puddletown Road, between Bovington and Wareham. From the Wareham direction, the small car park is on the right (north side of the road) 50m or so after the right turning signposted East Dorset Golf Club and Hyde.
Contact us
Environmental designation
Ramsar
Site of Special Scientific Interest (SSSI)
SPA
Special Areas of Conservation (SAC)
Higher Hyde Heath Nature Reserve Map
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-554,803,813,220,851,600 | Biological Sampling
Last updated:27 June 2014
The abundance of marine animals and their distribution is determined by a range of physical and biological factors such as:
• food quantity
• temperature
• salinity
• available space and
• broader-scale ocean currents.
To understand which benthic animals live in a variety of habitat conditions, ecologists at Geoscience Australia use a range of methods to sample the seafloor. These methods depend on both the size of the animals of interest and the habitat they occupy. Animals on the seafloor can be classified into two broad groups: epifauna and infauna.
Epifauna live on the surface of the seafloor and include mobile animals like bottom-dwelling fish, seastars, and molluscs as well as sessile animals like sponges, corals, and bryozoans. We use photography and video to observe these animals and collect them using sleds and dredges.
Much of the seafloor is composed of sand or mud that forms a three dimensional habitat that is home to huge numbers of small animals living in burrows and moving between the sediment grains. These infauna animals are crucial for the benthic ecosystem as they process dead plants and animals, circulate oxygen through the sediment, and act as food for bottom-feeding animals such as fish. Grabs and boxcores are used to sample both the seafloor sediments and their infauna. The animals are then carefully separated from the surrounding sediment by washing the sample through sieves with holes only 0.5 mm wide. Often thousands of small and microscopic animals are retained. Animals collected in the field are preserved for transport back to the laboratory where they are formally identified and counted under a microscope. Data from this work are then analysed to examine what types of animals live in different sediment types and how environmental conditions affect the distribution and abundance of marine life.
Zooplankton are animals that live suspended in the water. Benthic animals are linked to zooplankton through food webs, shared nutrients, and planktonic early life stages. Geoscience Australia is developing and applying new methods to sample zooplankton in order to investigate their links with benthic animals and how those links may be used in surrogacy research. Geoscience Australia has designed the Mounted Assembly for Planktobenthic Sampling (MAPS) and deployed it recently to achieve one of the first simultaneous collections of benthic animals and zooplankton. | {
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9,142,265,293,523,583,000 | Navigation Links
Largest study of human 'interactome' reveals a novel way
Discoveries made during the first large-scale analysis of interactions between proteins in our cells hold promise for identifying new genes involved in genetic diseases, according to researchers at Johns Hopkins and the Institute of Bioinformatics (IOB) in Bangalore.
The findings, reported in the March issue of Nature Genetics, were made using a database of more than 25,000 protein-protein interactions compiled by the Hopkins-IOB team. The result is believed to be the most detailed human "interactome" yet describing the interplay of proteins that occur in cells during health and disease.
"Genes are important because they are the blueprints for proteins, but proteins are where the action is in human life and health," says Akhilesh Pandey, M.D., Ph.D., an assistant professor at the Institute of Genetic Medicine and the departments of Biological Chemistry, Oncology and Pathology at The Johns Hopkins University School of Medicine. "This ability to find links between sets of proteins involved in different genetic disorders offers a novel approach for more rapidly identifying new candidate genes involved in human diseases," he says.
The analysis included interactions among 1,077 genes coding for proteins linked to 3,133 diseases, the researchers report. Significantly, it showed that proteins encoded by genes that are mutated in inherited disorders were likely to interact with proteins already known to cause similar disorders. In addition, the researchers disproved the long-held belief among scientists that the relative importance of a specific protein is always reflected by the number of other proteins it interacts with in the cell.
According to Pandey, the team's comparison of almost 25,000 human, 16,000 yeast, 5,500 worm, and 25,000 fly protein-protein interactions showed that, among these more than 70,000 links, only 16 were common to all four species.
Researchers say this low level of interactome overlap among species w as surprising. It showed that current rapid-testing methods for identifying protein interactions are likely to miss true interactions.
Much of the Hopkins-Bangalore work was based on information compiled in the Human Protein Reference Database (HPRD), a repository of information on protein-protein interactions collected from the published literature and stored in a format suitable for rapid study and comparison with other animal cells. HPRD was developed by the IOB and the Pandey laboratory.
"Using HPRD and several other databases, we have been able to develop a gold mine of new information for researchers seeking new ways of finding candidate genes involved in genetic diseases," Pandey says. "And our demonstration that a protein's importance is not based on the number of interactions it has with other proteins is an important conceptual breakthrough. It eliminates a blind alley that could mislead researchers investigating the roles of specific proteins in the cell."
Pandey is the chief scientific advisor to the IOB and senior author of the Nature Genetics article. The team's conceptual advance was made by comparing human data with 6,014 genes in yeast and 2,284 genes in mice whose effect on survival was known, according to Pandey. "Our much larger database on genes and proteins gave us the information to set the record straight on how to measure a protein's importance," he says.
Using this kind of comprehensive comparison of information about human and other organisms allowed Pandey's group to identify 36 previously unknown protein-protein interactions, nine of which were tested in the laboratory to verify what the analysis suggested. "We proved they were valid," Pandey says. "By linking computerized sleuthing to laboratory experiments to confirm those findings, we expect to be able to eventually fill in many blanks in human protein-protein interactions."
All the analyses were primarily carried out at the IOB, a nonprofit research institute founded by Pandey in May 2002. The Human Protein Reference Database was developed with funding from the National Institutes of Health and the Institute of Bioinformatics. Pandey serves as chief scientific advisor to the Institute of Bioinformatics. He is entitled to a share of licensing fees paid to The Johns Hopkins University by commercial entities for use of the database. The terms of these arrangements are being managed by The Johns Hopkins University in accordance with its conflict of interest policies.
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Source:Johns Hopkins Medical Institutions
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-1,596,918,605,745,503,500 | Precipitation over the rainforest -- sabah_aerial_0890
Image ID: sabah_aerial_0890 | Date photographed: 2012-Nov-07
Tags: malaysia, rainforests, n4 36.041 e116 57.251, borneo, borneo rainforest, asian rainforests, malaysian rainforests
Photographer: Rhett A. Butler | Camera:
Country: Malaysia Location: Sabah
Advanced search (by country, tag, country-tag combination, etc)
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5,218,022,066,953,785,000 | University of Lethbridge » uNews »
Hontela contributes to IET team
A University of Lethbridge ecotoxicology expert is part of a new province-wide research institute dedicated to examining the impact of harmful chemicals in our environment.
Dr. Alice Hontela, a U of L biological sciences professor and Canada Research Chair in ecotoxicology, is among the researchers who make up the Institute of Environmental Toxicology (IET). Housed at the University of Calgary, the IET is a multidisciplinary team that will include biologists and engineers from the universities of Lethbridge, Calgary and Alberta, as well as scientists in municipal and federal laboratories. Its mandate is to develop new and improved technologies for effective risk assessment and remediation of contaminated sites.
“Our research will help provide decision makers and the public with scientific information on the nature and magnitude of environmental contaminants, and offer some solutions to address this important global challenge,” says Hontela.
Among other research collaborations, Hontela is part of MITHE (Metals in The Human Environment) strategic network and the Alberta Institute for Water Research. She says climate change and increasing industrial activity (especially farming and mining) are not only affecting the volume of available water, but the concentration of pollutants that exist in the water.
Through lab and fieldwork, Hontela studies the endocrine systems of different fish to determine the effects of certain pollutants. She explains that some chemicals are safe at low doses (like selenium, which our bodies need), but toxic at higher concentrations.
Since pollutants, like pharmaceuticals, are difficult or impossible to remove with filtration, the health of both human and aquatic species requires keeping these chemicals at non-toxic levels.
“Alberta is a province that is changing very quickly, and we have to find new ways to deal with our water and our pollutants,” says Dr. Hamid Habibi, professor in the Department of Biological Sciences at the University of Calgary and the IET director.
An environment free of harmful chemicals is the group’s top priority. IET will study the nature, fate and persistence of chemicals in the environment as well as the remediation of terrestrial and aquatic contamination activities.
Habibi says that the causes of contamination are wide ranging and could include pollutants released from wastewater treatment plants and landfills, such as plasticizers, surfactants, flame-retardants, drugs, personal care and industrial products. In addition, there are heavy metals or other toxins resulting from mining activities and electronics consumerism, agricultural runoff, oil sands operations and tailings ponds.
For a full look at the January issue of the Legend, follow this link.
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-1,583,008,732,279,381,500 |
David M. Wright
There are 1 included publications by David M. Wright :
TitleDateViewsBrief Description
Rapid and improved assay of surfactins from Bacillus subtilis, 203R via UPLC-ESI-MS 2018 304 To better bridge research with commercialization, this project sought to develop an improved analytical method for the assay of biosurfactants known as surfactins from Bacillus subtilis. We sought to compare levels of production from various strains ... | {
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-8,817,351,557,289,025,000 | Introduction to Cell Biology, Cells, Cell Theory Lesson PowerPoint
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-445,404,064,287,737,500 | Advertisement
A Fluctuating Reality
A Fluctuating Reality Accused of fraud, Anders Pape Möller has traveled from superstar evolutionary biologist to pariah. By Brendan Borrell ARTICLE EXTRAS Origin of a Controversy Timeline: From Superstar to Pariah One day in the early spring of 1993, Richard Palmer received a paper by a Danish ornithologist, Anders Pape Möller. Palmer, an associate editor at Evolution, was impressed by the paper, but he was troubled by one of Möller's key st
By | January 1, 2007
A Fluctuating Reality
Accused of fraud, Anders Pape Möller has traveled from superstar evolutionary biologist to pariah.
By Brendan Borrell
One day in the early spring of 1993, Richard Palmer received a paper by a Danish ornithologist, Anders Pape Möller. Palmer, an associate editor at Evolution, was impressed by the paper, but he was troubled by one of Möller's key statistics.
Although he had met Möller only once, Palmer was familiar with his work. Both were fascinated by the promise of fluctuating asymmetry, the subject of the paper in question. "If you measure the right and left sides of the body very precisely, they're never exact mirror images," explains Palmer, an evolutionary biologist at the University of Alberta in Edmonton. "Those differences are random, and what they tell you is the inability of the right side of the body to produce an exact mirror of the left."
Möller's paper claimed that asymmetry in the tail feathers of the barn swallow was passed from fathers to their sons; in other words, it was heritable. But Palmer pointed out in a three-page letter to Möller that the statistical significance of his findings hinged upon a single outlying data point, and therefore "it would be more prudent to present the data, indicate the sensitivity of the statistical result to a single point, and conclude that it is not possible to say much about the heritability of asymmetry with the present data." Instead of addressing Palmer's concerns and those of the two reviewers, however, Palmer says he felt that Möller was just trying to make those concerns go away. Möller ultimately softened the language of the paper and Palmer accepted it for publication, although he says, "I was left with the sense that it was more important for him to get the paper published than to be correct."
Palmer wasn't alone. Evolutionary biologist Bob Montgomerie of Queens College says it's no secret that Möller bickers with editors and referees. As a frequent reviewer of Möller's papers, Montgomerie found himself endlessly pointing out mistakes, but "the stuff was getting published anyway."
PHOTO BY TIM MOUSSEAU Anders Pape Möller measures birds at a field site in the Ukraine in 2005. Möller and Tim Mousseau have been working on a project to investigate the effect of the Chernobyl disaster on biodiversity.
Meanwhile, a handful of Möller's colleagues had begun distancing themselves from him. His collegial relationship with evolutionary biologist Andrew Pomiankowski of University College London deteriorated after a dispute over one of their papers. Adrian Thomas, an ornithologist at the University of Oxford, stopped replying to Möller's E-mails regarding a proposed collaboration. Rumors began circulating about the ecologist, including one back-of-the-envelope calculation that retraced his putative bicycle route at his field research site using the sampling methodology described in concurrent studies. The velocities required an athlete of Olympic caliber.
These suspicions would move into the pages of journals, and eventually into a full-fledged investigation that cast serious doubt on one of Möller's papers. In 2005, Möller's bird-banding permit was revoked, effectively ending his 34-year study of barn swallows. "I've slept badly for five years, now," says Möller via the phone from his lab at the Pierre and Marie Curie University in Paris. "I don't think I have done anything wrong." He says his students have been harassed, his collaborators have been discouraged from working with him, and his family has suffered. His friend, Tim Mousseau, a biologist at the University of South Carolina, has seen firsthand how the investigations have affected him. "I think it was very hurtful for somebody who has dedicated their entire life to the pursuit of knowledge," he says, adding: "The only recognition he wants is for his science."
A FARMER'S BOY
Möller was born in the town of N?rresundby on the day after Christmas in 1953. N?rresundby lies in the peatlands of Denmark's sparsely populated Jutland peninsula and is the site of Lindholm H?je, a major Viking burial ground dating back more than a thousand years. While his ancestors took to the sea, Möller took to the land: "I was a farmer's boy."
During his youth, he tended to his father's cows, sheep, and chickens, and, when he had the chance, he watched birds. In the fall of 1969, 15-year-old Möller visited Thorkil Duch, an electrician and an amateur naturalist in the area, who advised him to keep a notebook of his observations. Duch also taught him to capture birds and wrap identifying bands around their legs so that Möller could keep track not just of species but also individuals. Möller returned home and started banding the barn swallow, a slight, nimble bird that would launch his scientific career.
Four years and untold notebooks later, Möller published his first scientific article on barn swallows in a Danish bird journal. He continued to publish throughout high school but was advised not to pursue a career in biology. "I was told there were so few positions that it would never pay off," he says. He went into biology anyway, and was accepted to a doctoral program at the University of Arhus. There, he quickly distinguished himself as a skilled ornithologist and a diligent worker. He wrote modest papers, focusing on mundane but telling details on the lives of common birds: when crows forage, how magpies die, and where blackbirds lay their eggs.
"I was left with the sense that it was more important for him to get the paper published than to be correct."
- Richard Palmer
Shortly after receiving his doctorate in 1985, he was publishing 20 to 30 papers a year in international journals: Behavioral Ecology and Sociobiology, Animal Behavior, Evolution, and Oikos, and in 2002 was selected as an ISI Highly Cited researcher in the field of ecology and the environment. He has now published nearly 600 papers. Dolph Schluter, another former editor of Evolution, says, "He's not just prolific. He's good. He's drawn comparisons [and] pointed to relationships that people will be digging through for years."
Part of Möller's success stemmed from his ability to forge productive collaborations. A list of his coauthors is a who's who in the field of behavioral ecology, and he was as likely to collaborate with a top scientist as with a provincial one.
When asked about his tremendous output, Möller laughs nervously and attributes it to his life on the farm: "You had to work hard to earn your dinner."
A FIELD TAKES OFF
Palmer, who published a review of asymmetry in Science in 2004, describes the early pioneers in the field with reverence: Lee Van Valen was "brilliant" and Kenneth Mather wrote "wonderful" papers. Articles on the topic had been trickling in since the 1940s, but the field really took off in the early 1990s thanks to Möller. "Without a doubt," Palmer says, "you can trace the spectacular popularity in this whole subject area to one paper Möller wrote on barn swallows."
Möller had previously shown that longer tails exhibited greater symmetry than shorter tails, a finding which led him to postulate that symmetry could be an indicator of "good genes." Möller's talent, Pomiankowski says, "is taking theoretical ideas and seeing ways they can be tested with data." So Möller promptly modified the length and asymmetry of the birds' tail feathers and found that females preferred the most symmetrical males (see sidebar). A paper, "Female Swallow Preference for Symmetrical Male Sexual Ornaments," was published in Nature in 1992 and was immediately touted by media outlets around the world: symmetry equals attractiveness.
Scientists were skeptical. "The results were too amazing to believe at face value, which was partly what made us look so closely at the paper," says evolutionary biologist Gerald Wilkinson at the University of Maryland, who criticized the study in a published note to Nature. He and ornithologist Gerald Borgia had noticed inconsistencies with error bars on graphs and doubted the paper's conclusions. Möller published a response to their criticisms, but as Wilkinson recalls, "The only way we could reconcile what he said is if his figures had been in error, if they had been crafted improperly."
HERITABLE ASYMMETRY?
In 1993, despite the doubts, evolutionary geneticist Therese Markow invited Palmer and Möller to a conference she organized at the Mission Palms Hotel in Tempe, Ariz. During the conference, Möller first suggested that asymmetry was heritable. This idea is a precondition for his "good genes" theory of sexual selection to apply to his barn swallows: If symmetric tails were not heritable, then they could not have evolved under sexual selection. "A rule of thumb is that everything is heritable," says Möller. "Some things have high heritability and some have a low heritability. This is one of the traits that has a low heritability, but it's very interesting."
Möller mentioned several important studies that demonstrated heritability, but the other attendants insisted that there were none. (Palmer agrees that some evidence exists for the heritability of asymmetry, but he says that it is one of the "squishier" connections.) Möller and Randy Thornhill, who was also at the meeting, set out to prove them wrong by performing a meta-analysis of the relationship between asymmetry and heritability. Thornhill, a professor at the University of New Mexico, Albuquerque, and coauthor of the controversial book Natural History of Rape, had been accused of sloppy science in the past. Palmer says he puts the two "in the same basket."
"I've slept badly for five years, now. I don't think I have done anything wrong."
- Anders Möller
Palmer rejected the manuscript at Evolution after receiving two "vitriolic" reviews that raised serious questions about its quality. Möller and Thornhill stood by their conclusions, and eventually the paper landed at a less prominent journal, Journal of Evolutionary Biology. The editor there sensed the brewing controversy and, in an unorthodox move, invited seven commentaries to be published alongside the original article in 1997.
The overall tone of these responses ranged from accusations of sloppiness to hyperbole to outright dishonesty. One set of authors suggested that Möller and Thornhill had a hidden agenda in analyzing their data: supporting their "good genes" model of sexual selection. Pomiankowski, who wrote a gentler response to the paper, says, "I was privy to earlier versions of his analysis, and the numbers kept on changing." In their reply, Möller and Thornhill deny a hidden agenda, adding that "there is a real danger to a scientific field when established workers in the field view their colleagues as competitors and use innuendos and direct claims of malpractice to try to get an edge." If Möller and Thornhill really thought they were fooling anyone, they were only fooling themselves.
THE FINAL STRAW
Then, in 1998, Möller published his 33rd paper in the Danish ecological journal Oikos, describing a relationship between asymmetry in oak leaves and damage caused by plant-eating insects. A year later, Oikos editor-in-chief Nils Malmer received an E-mail from Jorgen Rab?l, a former professor in Möller's lab at the University of Copenhagen, who suggested that the data had been fabricated. Möller was shocked. "I had saved all these bloody leaves from these trees," he recalls. "I thought perhaps there was something wrong with these measurements." He went back to his crackling leaf samples and remeasured them. He soon realized that the new data failed to support the conclusions in the Oikos paper. He felt humiliated and did what he and Malmer agreed was the only honorable response: He published a retraction.
That could have been the end of it. But to Möller's dismay, Rab?l brought the case before the Danish Committee on Scientific Dishonesty in 2001. Rab?l presented the committee with files he had obtained from Möller's technician, and the committee then requested Möller's own data files. Möller delayed for months, insisting that the raw data had been stolen along with his laptop in 1996. Instead, he sent the committee a transformed data set that served as the basis for the paper's three tables. The committee noted inconsistencies in even these files and ruled in 2003: "Neither the raw data kept at the University of Copenhagen nor the data forwarded by the defendant could have generated the results that emerged from the article."
Möller insists that the investigation did not prove his guilt but was instead a character assassination. Indeed, Rab?l had been fired after Möller complained of his lack of productivity, and Möller maintains that the accusations were part of Rab?l's revenge. Möller notes that a second investigation, conducted by his home institution, the National Center for Scientific Research (CNRS) in Paris, did not find him guilty of intentionally committing fraud. But even that verdict states that the committee was "lacking the material evidence necessary to establish innocence."
THE AFTERMATH
Möller still publishes at a healthy pace, although he says his manuscripts are rejected twice as frequently as before the investigation. "He's under the microscope," says former Evolution editor Schluter at the University of British Columbia. Yet a look at his recent papers shows that while he is keen on citing his own work, he rarely cites opposing views, perhaps hoping, as Palmer remarks, that they'll just "go away."
Perhaps in response, scientists remain critical and even unkind to Möller. In 2000, Palmer published an unusual essay in the newsletter for the International Society of Behavioral Ecology. It was a fable concerning the fictitious Traumweber brothers, Andy and Randy, expert tailors in the "remote kingdom of Gl?cklichtal, nestled high in the European Alps." Palmer wrote that the maestro of Gl?cklichtal's symphony noticed that audiences "seemed pleased with performances conducted in the Traumweber tuxedo, but dissatisfied when he performed in his imported tuxedo." After careful investigation, "Andy Traumweber discovered the imported jacket was less precisely made, most particularly in the tails: one was distinctly longer than the other." The title of the piece, "The Emperor's Codpiece," came from its final coup:
According to a palace informer, the Emperor was particularly anxious about his imperial private parts, which he felt were so asymmetrical that they deviated too far from the norm. Fortunately, the Traumweber brothers were able to allay his fears with a profound revelation: In certain very special cases, increased expression of a predictable asymmetry actually signals increased fitness, and one of those cases is testicles (Möller 1994), at least if men are like birds. That's why they subsequently fashioned the Emperor's codpiece to enhance his already conspicuous asymmetry.
The president of the society, Nick Davies, issued an ambivalent apology in the subsequent newsletter.
"Results were too amazing to believe at face value, which was partly what made us look so closely at the paper."
- Gerald Wilkinson
In a devastating book review of Asymmetry, DevelopmentalStability, and Evolution, evolutionary geneticist David Houle at Florida State University, wrote that Möller and his coauthor John Swaddle at the College of William and Mary "repeat the original conclusions of Möller and Thornhill's (1997) meta-analysis of the heritability of asymmetry down to the wildly inflated estimate of average heritability. Although they do address some of the criticisms of others, these are, in effect, dismissed as technical points that do not affect the overall conclusions."
In closing his review, Houle widens his scope to include the gullible souls who jumped aboard the fluctuating asymmetry bandwagon in the 1990s as well as all scientists who succumb too easily to the enthusiasm accompanying new ideas. "We have little choice," writes Houle, "but to seek inspiration from gurus of the newest ideas; sometimes they turn out to be partially right. However, we should never believe them without a struggle. If an idea seems too good to be true, it is probably not true."
These days, Möller's most vocal defender seems to be Mousseau. "I like Rich [Palmer] a lot," Mousseau says, "He's a friend of mine, but he's quite emotional and somewhat irrational in his stance: he just doesn't like Möller." Palmer privately wrote Mousseau and cautioned him not to be so cavalier in defending his colleague. Mousseau, in turn, wrote letters to both Nature and Science with more than 20 coauthors, defended Möller on discussion boards, and started a petition to give Möller back his bird-banding permit.
Pomiankowski says Möller is in the "limbo land" in which many scientists investigated for fraud find themselves. "I find it an unsatisfactory situation to be in, but that's where we are. I would much prefer that he was properly absolved for what happened or found properly guilty." He'd rather know the truth now. "It's very hard to understand what motivates another person," says Pomiankowski. "You can concoct an explanation about why things go wrong, but who knows?"
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Comments
Avatar of: David Herz
David Herz
Posts: 1
January 9, 2007
This well written piece is ample evidence, not that we need it, that even the highly educated, the dedicated, the above all suspicion can behave like vindictive children...except that they are adults, and I am not referring to Moller...In such a vindictive and emotionally wrought climate it is highly unlikely that the so called objective scientific spirit can wend its way to something approaching equity...the myth of the two tailors is evidence of great immaturity of spirit, misuse of power, and simple disrespect. \nthank you for allowing me to comment.
Avatar of: Charles Cortes
Charles Cortes
Posts: 1
January 9, 2007
I agree with Herz, the spirit of the article is just that "payback", unfortunately science is herd like. However, great minds excude ubiquitious pride which when fosterd in the minds of men are very often inflated along with egos.
Avatar of: Hugh Fletcher
Hugh Fletcher
Posts: 44
January 10, 2007
Scientific publications are full of results that literally do not add up, either through fraud or incompetence. In physical sciences where experiments are repeatable, this is usually discovered if it matters. Otherwise it just gets left, and the word goes around about being careful if you work with X. Can anyone honestly produce a worthwhile scientific paper every 18 days? I have been collecting papers containing meaningless rersults for decades. My favourite swallow tale suggested that cutting 20mm of the males? tails would increase their velocity from about 6 or 7m per second to around 15m/s. This is aerodynamically ridiculous, either the tip of the tale accounts for 75% of the drag, or the swallows were now expending 4 times the energy flying (the square of velocity). Do males normaly fly half as fast as females? The explanation was that they were not before and after tail cutting of each bird, they were different swallows flying in different places, and one or two of the fastest birds happened to be included in a particular group of 4. This, together with the surprising conclusion that the extra speed was bad for the birds, was published by Evans in Nature ( 394 p 233) and elicited a response from Anders Moller as to the exact selection causing these effects. Perhaps the enthusiasm of rank and file evolutionary biologists is not often matched by their numerical skills.
Avatar of: Larry Pinkerton
Larry Pinkerton
Posts: 1
January 10, 2007
As a layman I am comforted when scientists are held under the glaring light of peer review --it is why I love and support science --warts and all.
Avatar of: Michael Morris
Michael Morris
Posts: 3
January 18, 2007
Palmer, as associate editor of the respected journal Evolution, seems to have discharged his editorial duties appropriately with respect to Moller's papers. However, the writing of his mythical tale mocking Moller's work was immature and unprofessional. This is no way to deal with the serious issue of potential fraud - and Palmer should have known that.\n\nThis was an excellent article.
Avatar of: Michael Morris
Michael Morris
Posts: 3
January 18, 2007
I might add that Palmer's tale serves to deepen, rather than shed intelligent light, on the issue of scientific integrity in all its forms.\n\nSpecifically:\n(i) Can Palmer any longer be trusted as a peer reviewer of scientific articles if he resorts to scarcely veiled 'comedic' ridicule of other scientists in his field?\n(ii) What does this say about the Editor(s) of the International Society of Behavioral Ecology, who alowed Palmer's story to be published? A half-baked apology by the President, if that's what it was, seems insufficient.\n\nMore broadly, these incidents yet again bring into question the processes of peer review and editorship and cast another shadow (as highlighted, for example, by recent articles in The Scientist) over editorial integrity and professionalism.
January 18, 2007
With the many discussions and publications on the recent high-profile cases of scientific fraud, one starts to wonder if what we are witnessing is a new era of dishonesty or an accidental bout of fluctuating dishonesty.\nMy guess is that the incidences of scientific fraud will increase in the coming decade(s). While each case is founded in its own peculiar circumstances, the overall situation of raised stakes in science will contribute statistically to an overall increase in scientific fraud.\nThis ties in nicely with the debate on the number of scientists being trained:\n\nAre we training too many scientists? (The Scientist)\nAre There Too Many Postdocs? (Science)\nToo Many or Too Few? The Postdoc Production Policy Debate (Science)\n\nWith increased numbers of scientists and decreased funding, competition rises. Currently, every single scientist on this planet feels the pressure that he/she needs to become a science superstar in order to survive and obtain a position which will pay the bills. A superstar will only be born in a fashionable topic and thus these topics (largely controlled by a few science journals) are overrun, increasing competition further. Obviously, only very few will become science superstars. Consequently, the incentives of behaving fraudulently have never been larger than today. The number of fraud cases will inevitably follow this trend. Because of the huge incentives (getting a job and fame or landing on the streets in disgrace) I'm doubtful that any control measures can stop these parallel developments.\n\nHowever, reducing the incentives on the high end (less fame and prestige, less spin-off companies, patents and luxurious meetings sponsored by drug companies) and cushioning the low end (e.g. by capping grant size to increase overall grant number) will also decrease the number of fraud cases in science.\n\nIf you are a PostDoc with a family, your contract runs out in three months and every faculty position has 300 applicants, you really feel the temptation to fiddle a little with this one graph which will get you the publication you need to beat the other 299 in order to feed your family. Asking for honesty is probably rather ineffective in such a situation.\nThat's the much more common low-profile fraud which is probably not increasing but exploding as I'm typing this.\nThe reasons are clear. Are we going to do something about it?
Avatar of: John L. Morton
John L. Morton
Posts: 2
January 18, 2007
I feel that I must agree with Björn Brembs. In such a competitive environment these things are perhaps inevitable. The points made above about publication rate and reputation are also important. \n\nThere was a suggestion about a firing for a lack of productivity. In my cynical moments I've sometimes felt that a major part of the problem are some senior investigators who just want results, and as long as they are the right results never seem to question about where they came from, or how rapidly they had have been generated. As Dr Brembs pointed out, if feeding your family depends on this the temptation could be irresistable.\n\nA very good article, and very thought-provoking.\n\n
January 18, 2007
Perhaps not surprisingly, a news article just out in the journal Nature supports the "pressure cooker" hypothesis of Breeding Cheats (part of a feature on scientific misconduct).\nSo most likely, if the current funding situation continues we will see more and more cases of fraud.\n
Avatar of: Dave Peters
Dave Peters
Posts: 1
January 18, 2007
The article notes Houle's scathing review of the Moller and Swaddle book. However, no mention is made of the accusation of plagiarism made by Houle in that review.\n\nMoller apparently plagiarized from a manuscript that he was sent to review, without making attribution. A passage of 200 words was supposedly taken word for word from the then-unpublished work. An apology by Moller, who was sent the manuscript for peer review, was subsequently tendered on the book's web site. While this itself was not data fraud, it certainly amounts to misrepresentation of ideas and thus seems relevant to the discussion. \n\nI'm curious because I've read several accounts of the Moller fraud case and none of them refer to this incident. One also wonders what co-author Swaddle made of all this.
Avatar of: Jørgen Rabøl
Jørgen Rabøl
Posts: 2
January 24, 2007
I was the person accusing Anders Pape Møller for data fabrications in the Oikos paper from 1998 and also raised the case to the Danish Committees on Scientific Dishonesty.\n\nAfter about seven years of dispute with Møller I know for sure, that he will never admit that he did anything wrong. He seems unable to realize even himself, that he fabricates data whenever necessary and possible. If you are interested in the Danish decision and more details about the Møller-case, please refer to my web site: www.jorgenrabol.dk\n
Avatar of: Jette Andersen
Jette Andersen
Posts: 1
January 26, 2007
May I use this occasion to correct a few things? \nMøller got off scot free in the OIKOS case by blaiming the technician (me) and retracting the article on the grounds of "bad measurements". This collided with his thanking me profusely ("heroic task") - even twice, both in the acknowledgments and in the body of the article. Møller's act was the reason Rabøl brought the paper to the attention of the Danish Committees for Scientific Dishonesty. That was too low and cheap, not to mention untrue, for Rabøl to stomac. The animosity between the two arose from that fact, not from sour grapes, nor envy or vengefulness. Rabøl was simply discusted as is the author of this paper. Later Møller saw fit to publicly accuse me of alcoholism and substance abuse (again absolutely untrue), so the low level of arguments at least in our case was set very early and stemmed from Møller. Rabøl left Copenhagen University much later than Møller did. He was sixtyone at the time and simply retired. He is still affiliated to the department.\n
Avatar of: Robert Trivers
Robert Trivers
Posts: 1
February 5, 2007
Richard Palmer says he puts Randy Thornhill and Anders Moller ?in the same basket?, meaning equally unreliable. This over his statistical disagreements with a review paper they jointly co-authored. \n\nMoller and Thornhill are very different organisms but I would place them in the following basket. They are both brilliant biologists who have taught me a great deal of evolutionary biology. I can not say that about any of their detractors.\n\nOften overlooked in the so-called Moller scandal is the fact that Anders is?barring perhaps his work on oak leaves?always right. If he is inventing his data, he knows exactly how to invent it. Take his discovery that swallow tail asymmetry in males affects female choice in nature. Did he lead us astray?did a discipline gallop off in a bad direction. Not at all. His discovery has been confirmed in a wide range of birds, and experimentally (using leg bands) in two species. Did he lead us astray when he claimed that bumblebees prefer symmetrical flowers, which in turn are richer in nectar rewards? Not at all, there is a flourishing little discipline pursuing this subject now. Did he mislead us when he claimed that immune characters were related to asymmetry of tail feathers in swallows? Immune connections with symmetry are now routinely reported. Well, was Anders misleading us when he provided a superb review of these findings in 2006? It would be very hard to say ?yes? without doing the massive literature review that he did.\n\nIn short, you can take two polar views of Anders Moller. He is either a genius whose ?empirical? articles are best viewed as brilliant interpretations of what data would like if one bothered to collect it, or else a careful and thorough scientist, who brilliantly demonstrates the importance of key ideas, which later work confirms. Take your pick. In either case, he is a teacher worth paying close attention to.\n\nIn conversation and correspondence he has put me on to a steady stream of fascinating work that has nothing directly to do with his own, e.g. the importance of melanin as a factor protecting against infections (re skin color in humans) or a very novel test of self-deception he once suggested, following from the fact that hemispheric specialization in women is associated with their tendency to believe their left breasts are larger than their right. Has he ever misled me, sent me to work of doubtful quality, wasted my time with work that proved trivial etc? Not yet.\n\nThornhill has also failed to lead me astray. In fact, he has been one of the most reliable (and creative) guides to human sexual selection. Does anyone doubt his finding that women (not on the pill) prefer the smell of symmetrical men at the time of their ovulation? Then let them repeat the meticulous work that led to this finding. Do women off the pill who are paired with asymmetrical men fantasize at ovulation more often about extra-pair copulations? Well, let me put it this way. He and co-workers have now shown the same thing using MHC similarity between mates, and MHC similarity is unambiguous in its measurement (compared to fluctuating asymmetry).\n\nWhat about his critics? The first thing they do is throw the baby out with the bathwater. Statistical arguments over review papers are common, and it is inevitable that theoretical biases will affect the organization of data one reviews. But the point is that the first serious review was done by Thornhill and Moller. They did a tremendous service bringing together all the data (or most of it). If the presentation was skewed let those not busy actually doing original work, study the matter more carefully, with the aid of the references and analysis already provided to them. \n\nIt is worth bearing in mind, that Gregor Mendel?s great work in genetics was shown by R.A. Fisher in the mid-thirties to have a less than one in a million chance of occurring, without active fudging of the results by Mendel. Fisher?s statistical conclusion is beyond the dreams of Anders? critics, yet Mendel?s work is widely understood to have provided the foundation for the science of genetics. Did Mendel make up his results out of whole cloth? Of course not, he would not have come anywhere near the truth. Probably he did sufficient work to realize the underlying logic and then gathered data with strong expectations in mind. Either consciously or unconsciously data was organized to as to show the world what he thought he had discovered. \n\nRichard Palmer?s work I respect. He emphasizes methodological and statistical rigor and we all need that. However, his mind sometimes also has a negative cast to it?not only in obsessing over the possible transgressions of others?but also in tending to claim that associations are unlikely which later turn out to be commonplace. When my co-workers and I showed that human dance obeyed striking symmetry associations predicted from the work of Moller and Thornhill (using motion-capture cameras to create a relatively pure test of theory), Palmer thought he immediately spotted a hideous mistake. It looked like our subjects showed average levels of relative asymmetry of 10 to 30%. He slyly asked if perhaps our youngsters had an unusual degree of deformity. It soon became apparent that he had merely read the paper carelessly, that we added our nine asymmetries, instead of taking their average. Palmer?s kind of bias repeated day in and day out will drive a scientist further and further from the truth, as he struggles all the while to subject work he finds suspect to rigorous scrutiny.\n\nHoule is not in the same basket. His review of Moller and Swaddle?s book deserves to be published in the Annals of Psychiatry. It begins by thoroughly misrepresenting the history and logic of his own discipline, population genetics, and then goes downhill from there. In one wild moment, he claims that the only value of this important book, is in showing others how not to do science. Never mind that you could burn the entire book except for the bibliography and you would still have a treasure (or vice-versa, in which case you would have a wealth of useful and new ideas). Houle?s piste de resistance is the discovery that in online material accompanying the book, Moller and Swaddle fail to attribute 200 words in a row that they borrowed from someone else, a mistake to be sure, but not quite the end of the world.\n\nIn turn, corresponding with Houle is to enter into a strange world of accusation, denial and misrepresentation. When I sent him my dance paper, he asked me to resend it because my spam-detection machinery had blocked my outgoing attachment. Sounded unlikely on its face (indeed, the first such occurrence in my life) but a moment?s study of his message showed that Houle?s own university informed him that they blocked the attachment and he could retrieve it if he acted within 24 hours. Further correspondence produced displacement of his mistake on to others and additional bizzarities not worth describing.\n\nIn falling so easily into the roles of prosecutor and judge, both these organisms forget Jesus? lesson (among those of others) that we should ?judge not, lest we be judged, for with the judgment you pronounce shall you be judged?. A little bit less obsession with the possible failings of others and a little more with their own failings ought to produce positive results all the way around.\n\nFinally, I do think?as I have told Anders a long time ago?that he made a mistake not taking total personal blame for the oak leaf problem; that is always the better posture and would likely have put this matter to rest. But should he be crucified for choosing the route he did? Not in my book, basket or moral system. He is a brilliant biologist who has taught the world more than his fair share of the truth. He has his defects as do we all, but there is no way he can be right about a subject, time and time again, and be the charlatan his critics try to make him out to be.\n\n
Avatar of: Jorgen Rabol
Jorgen Rabol
Posts: 2
February 5, 2007
I read the comment by Robert Trivers and the hard core in his argumentation is that if you are a genius you are allowed - and perhaps even wellcomed - to be a cheater and data-fabricator.\n I am sorry to repeat myself but as far as I can figure out APM cheats whenever necessary and possible, i.e. the oak leave paper was not his one and only misconduct. On the my homepage www.jorgenrabol.dk I mention two further cases of suspected cheating/fabrications.
Avatar of: John Swaddle
John Swaddle
Posts: 1
February 16, 2008
Working with Moller once over 10 years ago still affects my career, as can be seen by the number of vitriolic comments posted here and elsewhere. It was a damaging experience where I ended-up feeling betrayed and caught in the crossfire of strong-minded individuals hell bent on destroying each other. To find out, after the book had been published, that Moller had copied a whole section of an unpublished manuscript by Geoff Clarke into the book (word-for-word) was heartbreaking. To then read an unrefereed book review accusing me of this plagiarism, when the author of the review knew this not to be true (he had talked to Geoff Clarke and got the whole scoop), was devastating. While I am unhappy about Moller?s actions, I also feel that unrefereed reviews and editorials are not the place to make formal accusations of misconduct. Surely accusations should go through at least as much peer review as the work that is being criticized. Journals should not resort to tabloid style reporting, no matter who is the target of criticism.
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6,883,423,802,687,137 | Purification of formaldehyde and formate dehydrogenases from pea seeds by affinity chromatography and S-formylglutathione as the intermediate of formaldehyde metabolism.
@article{Uotila1979PurificationOF,
title={Purification of formaldehyde and formate dehydrogenases from pea seeds by affinity chromatography and S-formylglutathione as the intermediate of formaldehyde metabolism.},
author={Lasse Uotila and Mirkka Koivusalo},
journal={Archives of biochemistry and biophysics},
year={1979},
volume={196 1},
pages={33-45}
}
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-7,035,553,812,691,332,000 | The Campus Chronicle
The Campus Chronicle,
an online newspaper
for University of West Georgia faculty and
staff, is the official
source for news and information to all members of the campus community.
I Am West Georgia
Barbara Ballentine
Barbara Ballentine is West Georgia.Job title: Assistant Professor, Biology Department.
What I really do: Try to balance an active research program with meaningful mentoring for all of my UWG students. My research involves understanding how ecology and evolution influence animal behavior – particularly songbirds. This means I get to play outside all spring and summer! I teach Ecology, Evolution, and Animal Behavior.
Years at UWG: Almost 2 (I will be starting my third year this fall).
Before UWG, I: Was a post-doc at the Smithsonian Institution where I worked in the Migratory Bird Center. Before moving to DC, I graduated from Duke University with a Ph.D. in biology. I got my start as a budding behavioral ecologist at the University of North Carolina at Chapel Hill.
One thing I would change about UWG: I would like an office with a window.
I was born in: The suburbs of Philadelphia.
Family members are: Kitty family: Stripey, Cookie, Rocco and Orange. People family: mom, sister and extended family live in Philadelphia suburbs; Jeremy Hyman, my significant other, lives in western NC (with Rocco and Orange). We like to think we have a country house (Sylva, NC) and town house (Carrollton, GA).
People describe me as: I guess it would depend on the perspective of the person. I’ve heard that students call me challenging but fair (I’m 100% ok with that). My colleagues will probably describe me as sociable and friendly. My close friends and family would probably describe me as funny and opinionated. Jeremy thinks I’m silly.
Most colleagues don’t know that: I am very silly.
Favorite things to do: I enjoy my work in the lab, field and classroom. I like music, comedy, eating delicious food and cooking food outside, spending time with friends and family, traveling to cool places to see cool birds with cool people.
Favorite quote: “Nothing in biology makes sense expect in the light of evolution.” - Theodosius Dobzhansky.
Proudest accomplishment so far: Taking the road less traveled to pursue my interests and passion in biology despite gnawing doubt and anxiety that, much to my surprise, lead to a very satisfying career that makes me very happy (most of the time).
Pet peeve: Willful ignorance.
Best advice ever received: “There is plenty of time to sleep when you are dead.” - Stephen Nowicki (my illustrious Ph.D. advisor)
Best advice for new faculty and staff: There is plenty of time to sleep when you are dead.
What I most want to contribute to students: Ultimately, I want to help students discover their passions and provide them with the tools and confidence to follow those passions. But at the very least I hope that I open students’ minds to a view of the world that they haven’t seen or considered before.
The book everyone should read: “On the Origin of Species,” Charles Darwin.
The movie everyone should watch: With the exception of silly comedies like “Shaun of the Dead,” “Napoleon Dynamite” or my all time favorite “Dazed and Confused,” I’m not a big Hollywood film watcher. I usually find them predictable, overly melodramatic and exploitative. But, I really like documentary films. So I think everyone should see “An Inconvenient Truth” by Davis Guggenheim.
The person, dead or alive, I’d most like to meet: Charles Darwin.
The place I’d love to travel to where I haven’t been yet: So many places! South America, Southeast Asia, Africa, the Grand Canyon….
Although you didn’t ask, I’d like to tell you anyway: Advice that I would offer to students – Be fearless; the only thing that is preventing you from doing what you want to do is fear.
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-5,454,190,153,312,161,000 | Share this post on:
This to nonproducing isolates Eupatilin cost expanding in ironlimited media was dependent on
This to nonproducing isolates developing in ironlimited media was dependent on the presence of receptor mutations. Although growth in both groups was stimulated, most likely because of the presence of smaller amounts of other iron chelators (like pyocyanin), isolates with no mutations PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25865820 showed a significantly greater enhance than these with mutations (t three.3, df 8.63, P 0.05) (Fig. 3A). The second assumption is that maintenance from the receptor is expensive and that retention, hence, indicates choice around the ability to uptake the ferripyoverdine complicated. Our data also support this, simply because the high mutation price of the receptor gene fpvA suggests that it can be a target of selection. The cost is unlikely toPNAS August 25, 205 vol. 2 no. 34 THE PYOVERDINE SYSTEMFeOuter membrane Inner membraneFeFpvA receptor FpvR antisigma aspect FpvI receptor sigma aspect PvdS pyoverdine sigma factor Pyoverdine Human transferrinPREDICTIONS Below LUNG ADAPTATIONPREDICTIONS Below SOCIAL ADAPTATIONPyoverdine presentPyoverdine absentPyoverdine presentPyoverdine absentFig. . The pyoverdine method. (Upper) The pyoverdine receptor FpvA spans the cell wall. In the absence of bound pyoverdine, the anti factor FpvR inhibits the expression of components FpvI and PvdS. Pyoverdine acquires iron from transferrin. When ferripyoverdine binds to the receptor, FpvR releases FpvI and PvdS. Release of FpvI initiates synthesis with the receptor FpvA, and PvdS initiates synthesis of pyoverdine (illustrated by arrows). (Decrease Left) If pyoverdine production is lost as an adaptation to the lung, receptor function also becomes redundant, irrespective of irrespective of whether pyoverdine produced by neighbors is out there. (Lower Appropriate) Nevertheless, if pyoverdine production is lost simply because of cheating, we expect to determine retention of receptor function within the presence of pyoverdine developed by other people and function only lost inside the absence of pyoverdine.Andersen et al.EVOLUTIONSEE COMMENTARY8 Mutations obs mutations exp(Fig. four). Added support for a price of even low receptor expression comes in the transmissible DK2 clone variety. In 973, DK2 acquired a mutation inside the factor fpvI gene, canceling upregulation of receptor expression in the presence of pyoverdine (Fig. ) as well as, lowering background receptor expression (eight, 27). This mutation is, nevertheless, followed by no fewer than 7 exceptional nonsynonymous fpvA mutations in nine independent lines across patients, suggesting that even restricted background expression of fpvA is expensive. Social Interactions Drive Selection on Pyoverdine Metabolism Offered that the receptor is pricey to sustain for isolates not generating pyoverdine, we then tested if the possibility of cheating selects for retention of receptor function. We found that the social environment, certainly, is key, because loss of function was dependent on loss of pyoverdine production by coinfecting isolates and not the focal isolate. Within the presence of pyoverdine producers, exactly where cheating is attainable, mutations within the receptor genes [fpvA, fpvR, and fpvI (named just after fecR and fecI in E. coli)] are very rare. On the other hand, within the absence of extrinsic pyoverdine, the number of mutations is drastically greater than expected for fpvA [P(X 33) pois(X; five.06) 0.00] plus the anti aspect fpvR [P(X six) pois (X; 2.) 0.05]. For fpvI, P(X 3) pois(X; 0.82) 0.05 (Fig. 3B). Longitudinal sampling of nonproducing isolates from 0 individuals further permitted us to measure latency to respond to adjustments in the social atmosphere (Mate.
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Satellite images show nearly all of Lake Okeechobee covered in algae
Lake Okeechobee on June 29, 2018 is mostly covered by a cyanobacteria bloom according to a satellite image from NOAA, derived from Copernicus Sentinel-3 data from EUMETSAT
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Satellite images show nearly all of Lake Okeechobee covered in algae | {
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-8,288,592,932,117,458,000 | Climate change and temperature dependent biogeography: oxygen limitation of thermal tolerance in animals
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hpoertner [ at ] awi-bremerhaven.de
Abstract
Recent years have shown a rise in mean global temperatures and a shift in the geographical distribution of ectothermic animals. For a cause and effect analysis the present paper discusses those physiological processes limiting thermal tolerance. The lower heat tolerance in metazoa compared to unicellular eukaryotes and bacteria suggests that a complex systemic rather than molecular process is limiting in metazoa.Whole animal aerobic scope appears as the first process limited at low and high temperatures linked to the progressively insufficient capacity of circulation and ventilation. Oxygen levels in body fluids may decrease reflecting excessive oxygen demand at high or insufficient aerobic capacity of mitochondria at low temperatures. Aerobic scope falls at temperatures beyond the thermal optimum and vanishes at low or high critical temperatures when transition to an anaerobic mitochondrial metabolism occurs. The adjustment of mitochondrial densities on top of parallel molecular or membrane adjustments appears crucial to maintain aerobic scope and to shift thermal tolerance.In conclusion, the capacity of oxygen delivery matches full aerobic scope only within the thermal optimum. At temperatures beyond only time limited survival is supported by residual aerobic scope, then anaerobic metabolism and finally molecular protection by heat shock proteins and antioxidative defence. In a cause and effect hierarchy the progressive increase in oxygen limitation at extreme temperatures may even enhance oxidative and denaturation stress. As a corollary, capacity limitations at a complex level of organisation, the oxygen delivery system, define thermal tolerance limits before molecular functions become disturbed.
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4148
DOI 10.1007/s001140100216
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Pörtner, H. O. (2001): Climate change and temperature dependent biogeography: oxygen limitation of thermal tolerance in animals , Naturwissenschaften, 88 , pp. 137-146 . doi: 10.1007/s001140100216
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-1,846,114,378,458,295,300 | PARD6G Products
Antibodies
PARD6G Antibody
PARD6G Antibody
NBP2-30757
Species: Hu
Applications: ICC/IF, IHC, IHC-P
Host: Rabbit Polyclonal
Proteins
PARD6G Recombinant Protein An ...
PARD6G Recombinant Protein Antigen
NBP2-30757PEP
Species: Hu
Applications: AC
Bioinformatics
Product By Gene ID 84552
Alternate Names
• PARD6G
• PAR-6 Gamma
• PAR6G
• Partitioning Defective 6 Homolog Gamma
• Par-6 Partitioning Defective 6 Homolog Gamma
• PAR-6 Gamma Protein
• PAR6gamma
• PAR-6G
• Par-6 Partitioning Defective 6 Homolog Gamma (C. Elegans)
• Par-6 (Partitioning Defective 6, C.Elegans) Homolog Gamma
• Par-6 Family Cell Polarity Regulator Gamma
Pathways for PARD6G
View related products by pathway and learn more about each of the pathways below.
Methylation
Dna Integration
Dna Repair
Dna Methylation
Localization
PTMs for PARD6G
Learn more about PTMs related to PARD6G.
Methylation
Bioinformatics Tool for PARD6G
Discover related pathways, diseases and genes to PARD6G. Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
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8,023,453,492,602,562,000 | The Mathematical Institute, University of Oxford, Eprints Archive
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Monro, H C and Gaffney, E. A. (2009) Modelling chemotherapy resistance in palliation and failed cure. Journal of Theoretical Biology, 257 (2). pp. 292-302.
Monro, H C and Gaffney, E. A. Modelling chemotherapy resistance in palliation and failed cure. Journal of Theoretical Biology, 257 (2). pp. 292-302.
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5,113,214,101,954,888,000 | English
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In Vitro Activity of Amphotericin B in Combination with Colistin against Fungi Responsible for Invasive Infections
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Bange, Gert
Max Planck Fellow Molecular Physiology of Microbes, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;
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Schwarz, P., Nikolskiy, I., Bidaud, A. L., Sommer, F., Bange, G., & Dannaoui, E. (2022). In Vitro Activity of Amphotericin B in Combination with Colistin against Fungi Responsible for Invasive Infections. J Fungi (Basel), 8(2). doi:10.3390/jof8020115.
Abstract
The in vitro interaction of amphotericin B in combination with colistin was evaluated against a total of 86 strains comprising of 47 Candida species (10 Candida albicans, 15 Candida auris, five Candida glabrata, three Candida kefyr, five Candida krusei, four Candida parapsilosis and five Candida tropicalis), 29 Aspergillus species (five Aspergillus flavus, 10 Aspergillus fumigatus, four Aspergillus nidulans, five Aspergillus niger, and five Aspergillus terreus), and 10 Rhizopus species (seven Rhizopus arrhizus, one Rhizopus delemar and two Rhizopus microsporus) strains. For the determination of the interaction, a microdilution checkerboard technique based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) reference method for antifungal susceptibility testing was used. Results of the checkerboard technique were evaluated by the fractional inhibitory concentration index (FICI) based on the Loewe additivity model for all isolates. Different inhibition endpoints were used to capture both the interaction at MIC and sub-MIC levels. Additionally, checkerboard technique results for Candida species were evaluated by response surface analysis based on the Bliss independence model. Against common Candida species, the combination was synergistic for 75% of the strains by FICI and for 66% of the strains by response surface analysis. For C. tropicalis, the interaction was antagonistic for three isolates by FICI, but antagonism was not confirmed by response surface analysis. Interestingly, synergistic and antagonistic FICIs were simultaneously present on checkboard microplates of all three strains. Against C. auris the combination was synergistic for 73% of the strains by response surface analysis and for 33% of the strains by FICI. This discrepancy could be related to the insensitivity of the FICI to detect weak interactions. Interaction for all other strains was indifferent. For Aspergillus and Rhizopus species combination exhibited only indifferent interactions against all tested strains. | {
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6,836,785,687,551,199,000 | Wetland and Aquatic Research Center
Connect
WARC scientists are located throughout the Southeast and Caribbean. Our two main centers are located in Gainesville, Florida, and Lafayette, Louisiana. We also have scientists based in offices in Baton Rouge, Louisiana; Davie, Florida; New Orleans, Louisiana; St. Petersburg, Florida; and St. John, U.S. Virgin Islands.
Contacts
Center Director (Supervisory Biologist)
Wetland and Aquatic Research Center
Phone: 352-264-3544
Deputy Director - Ecologist (Research)
USGS Wetland and Aquatic Research Center
Phone: 337-266-8647
Branch Chief -Supervisory Electronics Engineer
Wetland and Aquatic Research Center
Phone: 337-266-8644
Branch Chief — Imperiled and Invasive Aquatic Species
Wetland and Aquatic Research Center
Phone: 352-264-3515
Center Management Officer
Wetland and Aquatic Research Center
Phone: 337-266-8804 | {
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} | 57 | ff2a0dd8e83e1035faba2b053f3eae93 |
-3,375,460,052,878,276,600 | 학교소개
연구 성과
연구 성과 게시글의 상세 화면
제목 [논문] 이상호 교수님_ClpL is a functionally active tetradecameric AAA chaperonedistinct from hexameric/dodecameric ones
작성자 비임파성 장기 면역 연구센터 등록일 2021-01-26 조회수 184
ClpL is a functionally active tetradecameric AAA chaperonedistinct from hexameric/dodecameric ones
AAA+ (ATPases associated with diverse cellular activities) chaperones are involved in a plethora of cellular activities to ensure protein homeostasis. The function of AAA+ chaperones is mostly modulated by their hexameric/dodecameric quaternary structures. Here we report the structural and biochemical characterizations of a tetradecameric AAA+ chaperone, ClpL from Streptococcus pneumoniae. ClpL exists as a tetradecamer in solution in the presence of ATP. The cryo-EM structure of ClpL at 4.5 Å resolution reveals a striking tetradecameric arrangement. Solution structures of ClpL derived from small-angle X-ray scattering data suggest that the tetradecameric ClpL could assume a spiral conformation found in active hexameric/dodecameric AAA+ chaperone structures. Vertical positioning of the middle domain accounts for the head-to-head arrangement of two heptameric rings. Biochemical activity assays with site-directed mutagenesis confirmed the critical roles of residues both in the integrity of the tetradecameric arrangement and activities of ClpL. Non-conserved Q321 and R670 are crucial in the heptameric ring assembly of ClpL. These results establish that ClpL is a functionally active tetradecamer, clearly distinct from hexameric/ dodecameric AAA+ chaperones.
연구 성과 게시판의 이전글 다음글
이전글 [논문] 양시영 교수님_TRIM24-RIP3 axis perturbation accelerates osteoarthritis pathogenesis
다음글 [논문] 하상준 교수님_Tumor-infiltrating regulatory T cell accumulation in the tumor microenvironment is mediated by IL33/ST2...
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2,930,782,309,055,469,000 | 1. Florida Bay Dolphin Days
02:25
from Mac Stone / Added
Atlantic bottlenose dolphins dominate the shallow waters of Florida Bay. On calm days they ride the wake and bow of my boat jumping and spinning in the slipstream. This is a compilation of clips from outings within Everglades National Park and Dolphin Cove where I had the chance to get in the water and play with a pod of these incredible animals. I hope you enjoy. Video production and editing: Mac Stone Music: "The Ocean" by Sunny Day Real Estate Equipment: Canon 5dmkii with Canon 24-105mm lens and GoPro Hero2 for underwater sequences Special thanks to Adam Chasey for captaining the boat and Jessica Lundstrum, Emily Campbell, and Jessica Lilli for facilitating the swim. www.MacStonePhoto.com
+ More details
• Timelapse Experiments - Pine Island Campground, Everglades
00:27
from Kevin Huggins / Added
314 Plays / / 10 Comments
Visited Pine Island Campground in the Everglades with Gina, in my quest for more Milky Way footage, but the weather didn't cooperate. Oh well still liked some of the footage I got, so I incorporated it with some of the footage from the first visit. Thanks to Skywatcher (vimeo: user7686907) for the tip on creating a "darkframe" for the timelapse footage... was able to remove practically all of the hot/dead pixels in the footage... really great tip. Even though I'm really addicted to capturing Milky Way timelapse footage, I don't think I'll be visiting the Everglades until it gets alot cooler again (Nov/Dec) or I get a Hazmat suit... the mosquitoes are too brutal! I couldn't even compose the shots the way I wanted too cause the mosquitoes were attacking my face and hands incessantly! Thanks to Gina for being my grip assistant and mosquito killer :) Music: "One Million Miles" - ATB Location: Pine Island Campgrounds, Everglades National Park, Homestead, FL Main Gear: Canon 5DMKIIs and Rebel T3i Manfrotto Tripods and Fluid Heads Kessler Crane Stealth and Oracle Controller
+ More details
• Everglades Water
03:08
from UCONN Journalism Media / Added
73 Plays / / 0 Comments
Short video about the water resources and local pressures from individuals and corporations that threaten it Produced by Gwen Craig and Olivia Balsinger
+ More details
• Guardians of the Everglades Trailer
05:37
from Connie Bransilver / Added
34 Plays / / 0 Comments
5 minute Trailer for the in-production one hour documentary film to accompany the multi-media Guardians of the Everglades production. Nicholas Petrucci, Clyde Butcher and Connie Bransilver, artists, feature ten living Guardians of the Everglades. Director Sandy Cannon-Brown shines with this preview.
+ More details
• Human Destruction of the Florida Everglades
06:36
from Jessica Daly / Added
86 Plays / / 0 Comments
The range of ecosystems, wildlife, plant life and different endangered species that can be found in the Florida Everglades contribute to its importance and need for preservation. Along with these factors, the Everglades holds the number one source of existence for life in Southern Florida: water. The water that comes from the Everglades provides for more than 6 million people in South Florida. Despite this, humans continue to destroy these ecosystems and their wildlife, while polluting the water with Mercury and phosphates. Because of this, more than 50% of the original wetlands have been destroyed. My focus surrounds Tamiami Trail, a road built in the 1920s, which created an enormous barrier of water flow from the northern areas into Everglades National Park. Many people refuse to face the truth that if action is not immediately taken to save the Everglades, we will not only destroy many diverse and unique sets of species, we will in return destroy ourselves. Without water and life, we are nothing.
+ More details
• How to Dispatch a Python
00:55
from 3BL Media / Added
The Mohave Desert and the Everglades are not your typical destinations for spring break. Neither are the students who want to go there. For the fifth consecutive year, SCA and American Eagle Outfitters teamed up to provide eco-conscious spring breakers with the ultimate week-long educational and work experience. During the month of March, 120 college students from around the country descended upon two of our most environmentally challenged and imperiled habitats for an expense-paid week of hands-on conservation at the Everglades and Joshua Tree National Parks. Students helped rid the parks of destructive invasive plants, repair eroded hiking trails, assess wildlife health, and more. And while they aid threatened ecosystems, the students also gained new skills, environmental knowledge, and leadership capabilities. While at the Everglades, SCA members also came across an interesting "friend". Check out the Burmese Python they found as documented by videographer and ASB participant Patrick Canavan. You never know what you are going to find out there. Follow these future conservation leaders and get inspired! Read their blogs and view their photos at http://followme.thesca.org/
+ More details
• Timelapse Experiments - Milky Way Astro Timelapse Test
00:14
from Kevin Huggins / Added
483 Plays / / 3 Comments
My first attempt at timelapsing the Milky Way. Anthony and I decided to attempt it at Pine Island Campgrounds in Everglades National Park. The mosquitoes were out in force, but decently bearable when the breeze was blowing. We didn't stay long, hence the really short timelapses... next time we will do longer ones. I had never seen the milky way with my naked eye, so this was a lot of firsts for me... the Lyriad meteor shower was also this weekend and we did see quite a few. Only major issue we had with the cameras was the tonne of dead/hot pixels that showed up after all the long exposures :( As you can see if you look carefully... I still didn't catch all when I tried to remove most of them in Lightroom... there were over 100 of them in each of our 5DMKIIs. If anyone knows how to minimize their occurrences please let me know. Thanks. It was really alot of fun capturing the night sky like this... can't wait to do another... I just have to recover from all the mosquito bites!!! Music: "One Million Miles" - ATB Location: Pine Island Campgrounds, Everglades National Park, Homestead, FL Main Gear: Canon 5DMKIIs Manfrotto Tripods and Fluid Heads
+ More details
• 3D Literature Project
14:14
from Patty / Added
36 Plays / / 0 Comments
I finally have the file on my computer and I am uploading it to vimeo to keep it safe. :) This is my favorite memory from Mr. Horowitz's Ap Literature class! Senior year 2009-2010 Everglades high School.
+ More details
• Sunset on the Everglades
00:10
from Luis Rivas / Added
23 Plays / / 0 Comments
Sunset - Everglades Fl
+ More details
• The Italian job part 1
01:35
from Adam Silver / Added
60 Plays / / 0 Comments
Scouting for my good friends and clients from Italy for Maserati in Miami, Florida. Out in the Everglades & beyond!
+ More details
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-3,883,818,417,276,503,000 | Format
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Nat Chem Biol. 2014 Nov;10(11):963-8. doi: 10.1038/nchembio.1659. Epub 2014 Sep 28.
A roadmap for natural product discovery based on large-scale genomics and metabolomics.
Author information
1
Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
2
1] Department of Chemistry, Northwestern University, Evanston, Illinois, USA. [2] Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA. [3] Feinberg School of Medicine, Northwestern University, Evanston, Illinois, USA.
3
Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
4
Bacterial Foodborne Pathogens and Mycology Research, US Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, Illinois, USA.
5
1] Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [2] Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Abstract
Actinobacteria encode a wealth of natural product biosynthetic gene clusters, whose systematic study is complicated by numerous repetitive motifs. By combining several metrics, we developed a method for the global classification of these gene clusters into families (GCFs) and analyzed the biosynthetic capacity of Actinobacteria in 830 genome sequences, including 344 obtained for this project. The GCF network, comprising 11,422 gene clusters grouped into 4,122 GCFs, was validated in hundreds of strains by correlating confident mass spectrometric detection of known small molecules with the presence or absence of their established biosynthetic gene clusters. The method also linked previously unassigned GCFs to known natural products, an approach that will enable de novo, bioassay-free discovery of new natural products using large data sets. Extrapolation from the 830-genome data set reveals that Actinobacteria encode hundreds of thousands of future drug leads, and the strong correlation between phylogeny and GCFs frames a roadmap to efficiently access them.
PMID:
25262415
PMCID:
PMC4201863
DOI:
10.1038/nchembio.1659
[Indexed for MEDLINE]
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School Supplies > eBooks for Teachers > E-Lessons eBooks for Teachers > Specialized Cells Protect the Body from Harmful Substances (Resource Book Onl
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This unit about the human body encourages students to investigate how cells protect the body through two experiments: Peeking at Pathogens (observing dry/wet bread) & Skin Shield (observing broken/unbroken skin on apples). Includes information, visual (Microbes), directions, & response forms. (Find other units by searching 'Human Body 4-6')
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7,240,200,970,438,679,000 | School of Biomedical Sciences Publications - UQ eSpace http://espace.library.uq.edu.au/ The University of Queensland en Fez http://blogs.law.harvard.edu/tech/rss Architecture of the sperm whale forehead facilitates ramming combat http://espace.library.uq.edu.au/view/UQ:384192 2016-04-07T14:26:25Z Panagiotopoulou, Olga; Spyridis, Panagiotis; Mehari Abraha, Hyab; Carrier, David R.; Pataky, Todd C. Architecture of the sperm whale forehead facilitates ramming combat http://espace.library.uq.edu.au/view/UQ:384178 Finite element models, regional stress data and all probabilist data in relation the investigation of whether the architecture of the sperm whale forehead facilitates ramming combat. <br /> 2016-04-07T13:05:33Z Panagiotopoulou, Olga Architecture of the sperm whale forehead facilitates ramming combat http://espace.library.uq.edu.au/view/UQ:384151 2016-04-07T10:50:01Z Panagiotopoulou,Olga; Spyridis, Panagiotis; Mehari Abraha, Hyab; Carrier, David R.; Pataky, Todd C. Are avian eggshell colours effective intraspecific communication signals in the Muscicapoidea? A perceptual modelling approach http://espace.library.uq.edu.au/view/UQ:184442 Diversity in the colour and appearance of avian eggshells has been proposed to serve a variety of visual functions, including crypsis from predation, mimicry and discrimination in facultative and obligate brood parasitism, and sexually selected intraspecific signalling of the extent of maternal investment in the egg. Here, we apply a photoreceptor noise-limited colour opponent model of avian perception to assess a necessary corollary of any intraspecific signalling hypothesis, namely that individual birds are able to discriminate between colours of eggs in different conspecific clutches. Clutches from 46 species in the superfamily Muscicapoidea were measured at the Natural History Museum collection in Tring, UK. The results demonstrate that, for these particular species, most eggs are predicted not to be easily discriminable from those in other conspecific clutches in terms of the shells' background coloration. These findings are of fundamental concern to any signalling hypothesis that looks to explain the evolution of avian-visible egg colour polymorphism through selection at the intraspecific level. Importantly, future studies should combine both the proximate mechanisms and the ultimate functions of trait variability when testing hypotheses of the variability in eggshell appearance. 2009-10-02T11:12:42Z Cassey, P.; Ewen, J. G.; Marshall, N. J.; Vorobyev, M.; Blackburn, T. M.; Hauber, M. E. Are corals colorful? http://espace.library.uq.edu.au/view/UQ:80973 Using in situ spectrometry data and visual system modeling, we investigate whether the colors conferred to the reef-building corals by GFP-like proteins would look colorful not only to humans, but also to fish occupying different ecological niches on the reef. Some GFP-like proteins, most notably fluorescent greens and nonfluorescent chromoproteins, indeed generate intense color signals. An unexpected finding was that fluorescent proteins might also make corals appear less colorful to fish, counterbalancing the effect of absorption by the photosynthetic pigments of the endosymbiotic algae, which might be a form of protection against herbivores. We conclude that GFP-determined coloration of corals may be an important factor in visual ecology of the reef fishes. 2007-08-15T09:17:33Z Matz, Mikhail V.; Marshall, N. Justin; Vorobyev, Misha Are fruit colors adapted to consumer vision and birds equally efficient in detecting colorful signals? http://espace.library.uq.edu.au/view/UQ:127744 Reproduction in plants often requires animal vectors. Fruit and flower colors are traditionally viewed as an adaptation to facilitate detection for pollinators and seed dispersers. This long-standing hypothesis predicts that fruits are easier to detect against their own leaves compared with those of different species. We tested this hypothesis by analyzing the chromatic contrasts between 130 bird-dispersed fruits and their respective backgrounds according to avian vision. From a bird's view, fruits are not more contrasting to their own background than to those of other plant species. Fruit colors are therefore not adapted toward maximized conspicuousness for avian seed dispersers. However, secondary structures associated with fruit displays increase their contrasts. We used fruit colors to assess whether the ultraviolet and violet types of avian visual systems are equally efficient in detecting color signals. In bright light, the chromatic contrasts between fruit and background are stronger for ultraviolet vision. This advantage is due to the lesser overlap in spectral sensitivities of the blue and ultraviolet cones, which disappears in dim light conditions. We suggest that passerines with ultraviolet cones might primarily use epigamic signals that are less conspicuous to their avian predators (presumably with violet vision). Possible examples for such signals are carotenoid-based signals. 2008-02-18T16:03:17Z Schaefer, H. Martin; Schaefer, Veronika; Vorobyev, Misha A regenerative antioxidant protocol of vitamin E and α-lipoic acid ameliorates cardiovascular and metabolic changes in fructose-fed rats http://espace.library.uq.edu.au/view/UQ:244997 Type 2 diabetes is a major cause of cardiovascular disease.We have determined whether the metabolic and cardiovascular changes induced by a diet high in fructose in young adult male Wistar rats could be prevented or reversed by chronic intervention with natural antioxidants. We administered a regenerative antioxidant protocol using two natural compounds: α-lipoic acid together with vitamin E (α-tocopherol alone or a tocotrienol-rich fraction), given as either a prevention or reversal protocol in the food. These rats developed glucose intolerance, hypertension, and increased collagen deposition in the heart together with an increased ventricular stiffness. Treatment with a fixed combination of vitamin E (either α-tocopherol or tocotrienol-rich fraction, 0.84 g/kg food) and α-lipoic acid (1.6 g/kg food) normalized glucose tolerance, blood pressure, cardiac collagen deposition, and ventricular stiffness in both prevention and reversal protocols in these fructose-fed rats. These results suggest that adequate antioxidant therapy can both prevent and reverse the metabolic and cardiovascular damage in type 2 diabetes. 2011-08-05T12:07:29Z Patel, Jatin; Matnor, Nur Azim; Iyer, Abishek; Brown, Lindsay Are pioneer axons guided by regulatory gene expression domains in the zebrafish forebrain? High-resolution analysis of the patterning of the zebrafish brain during axon tract formation http://espace.library.uq.edu.au/view/UQ:59763 Although the principles of axon growth are well understood in vitro the mechanisms guiding axons in vivo are less clear. It has been postulated that growing axons in the vertebrate brain follow borders of neuroepithelial cells expressing specific regulatory genes. In the present study we reexamined this hypothesis by analysing the earliest growing axons in the forebrain of embryonic zebrafish. Confocal laser scanning microscopy was used to determine the spatiotemporal relationship between growing axons and the expression pattern of eight regulatory genes in zebrafish brain. Pioneer axons project either longitudinally or dorsoventrally to establish a scaffold of axon tracts during this developmental period. Each of the regulatory genes was expressed in stereotypical domains and the borders of some were oriented along dorsoventral and longitudinal planes. However, none of these borders clearly defined the trajectories of pioneer axons. In two cases axons coursed in proximity to the borders of shh and pax6, but only for a relatively short portion of their pathway. Only later growing axons were closely apposed to the borders of some gene expression domains. These results suggest that pioneer axons in the embryonic forebrain do not follow continuous pathways defined by the borders of regulatory gene expression domains, (C) 2000 Academic Press. 2007-08-14T16:02:31Z Hjorth, J. T.; Key, B. Are they using my feedback? The extent of students’ feedback use has a large impact on subsequent academic performance http://espace.library.uq.edu.au/view/UQ:387605 2016-05-27T13:26:01Z Zimbardi, Kirsten; Colthorpe, Kay; Dekker, Andrew; Engstrom, Craig; Bugarcic, Andrea; Worthy, Peter; Victor, Ruban; Chunduri, Prasad; Lluka, Lesley; Long, Phil A review of longnose skates Zearaja chilensis and Dipturus trachyderma (Rajiformes: Rajidae) http://espace.library.uq.edu.au/view/UQ:370426 2015-09-28T13:24:29Z Vargas-Caro, Carolina; Bustamante Diaz, Carlos; Lamilla, Julio; Bennett, Michael B. A review of the molecular basis of hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency http://espace.library.uq.edu.au/view/UQ:184150 Hypoxanthine-guanine phosphoribosyltransferase (HPRT, EC 2.4.2.8) is a purine salvage enzyme that catalyses the conversion of hypoxanthine and guanine to their respective mononucleotides. Partial deficiency of this enzyme can result in the overproduction of uric acid leading to a severe form of gout, whilst a virtual absence of HPRT activity causes the Lesch-Nyhan syndrome which is characterised by hyperuricaemia, mental retardation, choreoathetosis and compulsive self-mutilation. The HPRT-encoding gene is located on the X chromosome in the region q26–q27 and consists of nine exons and eight introns totalling 57 kb. This gene is transcribed to produce an mRNA of 1.6 kb, which contains a protein encoding region of 654 nucleotides. With the advent of increasingly refined techniques of molecular biology, it has been possible to study the HPRT gene of individuals with a deficiency in HPRT activity to determine the genetic basis of the enzyme deficiency. Many different mutations throughout the coding region have been described, but in the absence of precise information on the three-dimensional structure of the HPRT protein, it remains difficult to determine any consistent correlation between the structure and function of the enzyme. 2009-09-24T10:30:10Z Sculley, Donna G.; Dawson, Paul A.; Emmerson, Bryan T.; Gordon, Ross B. A review of the neural mechanisms of action and clinical efficiency of Riluzole in treating Amyotrophic Lateral Sclerosis: What have we learned in the last decade? http://espace.library.uq.edu.au/view/UQ:207199 2010-07-08T14:25:16Z Bellingham, Mark C. A review of vertebrate and invertebrate ocular filters http://espace.library.uq.edu.au/view/UQ:146828 2008-06-06T12:26:16Z Douglas, R. H.; Marshall, J. Are you calling me primitive? http://espace.library.uq.edu.au/view/UQ:77937 2007-08-15T07:24:21Z Schwab, I.R.; Collin, S. P. Argyrophilic grain disease http://espace.library.uq.edu.au/view/UQ:189284 2009-12-07T11:49:47Z Tolnay, M.; Ghebremedhin, Estifanos; Probst, Braak H. A rodent model of low- to moderate-dose ethanol consumption during pregnancy: Patterns of ethanol consumption and effects on fetal and offspring growth http://espace.library.uq.edu.au/view/UQ:268262 2012-02-24T12:10:21Z Probyn, Megan E.; Zanini, Simone; Ward, Leigh C.; Bertram, John F.; Moritz, Karen M. A role for cingulate pioneering axons in the development of the corpus callosum http://espace.library.uq.edu.au/view/UQ:109310 In many vertebrate and invertebrate systems, pioneering axons play a crucial role in establishing large axon tracts. Previous studies have addressed whether the first axons to cross the midline to from the corpus callosum arise from neurons in either the cingulate cortex (Koester and O'Leary [1994] J. Neurosci. 11:6608-6620) or the rostrolateral neocortex (Ozaki and Wahlsten [1998] J. Comp. Neurol. 400:197-206). However, these studies have not provided a consensus on which populations pioneer the corpus callosum. We have found that neurons within the cingulate cortex project axons that cross the midline and enter the contralateral hemisphere at E15.5. By using different carbocyanine dyes injected into either the cingulate cortex or the neocortex of the same brain, we found that cingulate axons crossed the midline before neocortical axons and projected into the contralateral cortex. Furthermore, the first neocortical axons to reach the midline crossed within the tract formed by these cingulate callosal axons, and appeared to fasciculate with them as they crossed the midline. These data indicate that axons from the cingulate cortex might pioneer a pathway for later arriving neocortical axons that form the corpus callosum. We also found that a small number of cingulate axons project to the septum as well as to the ipsilateral hippocampus via the fornix. In addition, we found that neurons in the cingulate cortex projected laterally to the rostrolateral neocortex at least 1 day before the neocortical axons reach the midline. Because the rostrolateral neocortex is the first neocortical region to develop, it sends the first neocortical axons to the midline to form the corpus callosum. We postulate that, together, both laterally and medially projecting cingulate axons may pioneer a path for the medially directed neocortical axons, thus helping to guide these axons toward and across the midline during the formation of the corpus callosum. (C) 2001 Wiley-Liss, Inc. 2007-09-19T16:24:40Z Rash, B. G.; Richards, L. J. A role for eosinophils in airway remodelling in asthma http://espace.library.uq.edu.au/view/UQ:233995 2011-03-09T14:29:54Z Kay, A. B.; Phipps, S.; Robinson, D. S. A role for Rho in smooth muscle phenotypic regulation http://espace.library.uq.edu.au/view/UQ:96591 The role of the small GTP-binding protein Rho in the process of smooth muscle cell (SMC) phenotypic modulation was investigated using cultured rabbit aortic SMCs. Both Rho transcription and Rho protein expression were high for the first 3 days of culture (contractile state cells), with expression decreasing after change to the synthetic state and peaking upon return to the contractile phenotype. Activation of Rho (indicated by translocation to the membrane) also peaked upon return to the contractile state and was low in synthetic state SMCs. Transient transfection of synthetic state rabbit SMCs with constitutively active Rho (val14rho) caused a dramatic decrease in cell size and reorganization of cytoskeletal proteins to resemble those of the contractile phenotype; alpha-actin and myosin adopted a tightly packed, highly organized arrangement, whereas vimentin localized to the immediate perinuclear region and focal adhesions were enlarged. Conversely, specific inhibition of endogenous Rho, by expression of C3 transferase, resulted in the complete loss of actin and myosin filaments without affecting the distribution of vimentin. Focal adhesions were reduced in number. Thus, Rho plays a key role in regulating SMC phenotypic expression. 2007-08-24T00:39:50Z Campbell, G. R.; Worth, N. F.; Rolfe, B. E. A role for Rho in smooth muscle phenotypic regulation http://espace.library.uq.edu.au/view/UQ:60742 The role of the small GTP-binding protein Rho in the process of smooth muscle cell (SMC) phenotypic modulation was investigated using cultured rabbit aortic SMCs. Both Rho transcription and Rho protein expression were high for the first 3 days of culture ("contractile" state cells), with expression decreasing after change to the "synthetic" state and peaking upon return to the contractile phenotype. Activation of Rho (indicated by translocation to the membrane) also peaked upon return to the contractile state and was low in synthetic state SMCs. Transient transfection of synthetic state rabbit SMCs with constitutively active Rho (vall4rho) caused a dramatic decrease in cell size and reorganization of cytoskeletal proteins to resemble those of the contractile phenotype; alpha-actin and myosin adopted a tightly packed, highly organized arrangement, whereas vimentin localized to the immediate perinuclear region and focal adhesions were enlarged. Conversely, specific inhibition of endogenous Rho, by expression of C3 transferase, resulted in the complete loss of actin and myosin filaments without affecting the distribution of vimentin. Focal adhesions were reduced in number. Thus, Rho plays a key role in regulating SMC phenotypic expression. 2007-08-14T16:45:39Z Worth, N. F.; Campbell, G. R.; Rolfe, B. E. A role for serotonin (5-HT) in hepatic stellate cell function and liver fibrosis http://espace.library.uq.edu.au/view/UQ:79378 2007-08-15T08:18:50Z Ruddell, Richard G.; Oakley, Fiona; Hussain, Ziafat; Yeung, Irene; Bryan-Lluka, Lesley J.; Ramm, Grant A.; Mann, Derek A. Arresting angiotensin type 1 receptors http://espace.library.uq.edu.au/view/UQ:160660 The type 1 angiotensin (AT1) receptor mediates the homeostatic and pathological actions of the peptide hormone, angiotensin II. With regard to the processes that activate and deactivate seven-transmembranespanning, G-protein-coupled receptors (GPCRs), AT1 receptors are among the most widely studied, serving as prototypes for GPCRs that bind and respond to peptide hormones. Arrestins are proteins that bind to activated and phosphorylated GPCRs, terminating initial signals emanating from these receptors, in addition to mediating receptor internalization. New aspects of arrestin function continue to emerge, such as their capacity to act as scaffolds to recruit regulatory and signaling molecules to increase the repertoire of receptor responses. Here, we examine the evidence that arrestins contribute to the signaling, deactivation and trafficking of AT1 receptors. 2009-01-14T10:49:39Z Thomas, Walter G.; Qian, Hongwei Arterially perfused eye model of uveitis http://espace.library.uq.edu.au/view/UQ:144301 2008-06-10T15:01:51Z Shiels, I. A.; Sanderson, S. D.; Taylor, S. M. Arterial wall renin-angiotensin-system and its role in hypertension http://espace.library.uq.edu.au/view/UQ:369658 2015-09-18T10:19:38Z Chen, Chen; Chen, Meng-Chin Arthropod Vaccines http://espace.library.uq.edu.au/view/UQ:143506 2008-06-10T14:23:01Z Lee, R.; Opdebeeck, J. P. Arylamine N-acetyltransferase 1: A novel drug target in cancer development http://espace.library.uq.edu.au/view/UQ:265638 2012-01-22T12:42:51Z Butcher, Neville J.; Minchin, Rodney F. Arylamine N-acetyltransferase 1 gene regulation by androgens requires a conserved heat shock element for heat shock factor-1 http://espace.library.uq.edu.au/view/UQ:205468 2010-05-23T00:05:35Z Butcher, Neville J.; Minchin, Rodney F. Arylamine N-acetyltransferase - from xenobiotic metabolising enzyme to regulation of transcription http://espace.library.uq.edu.au/view/UQ:41794 2007-08-13T14:38:16Z Minchin, Ron Arylamine N-Acetyltransferase Gene Polymorphism http://espace.library.uq.edu.au/view/UQ:72038 2007-08-14T12:29:30Z Butcher, N. J.; Minchin, R. F. Arylamine N-acetyltransferase I http://espace.library.uq.edu.au/view/UQ:130350 Arylamine N-acetyltransferase I (NAT1) is a phase II enzyme that acetylates a wide range of arylamine and hydrazine substrates. The NAT1 gene is located on chromosome 8 and shares homology to NAT genes found in most mammalian species. Gene expression occurs from at least two promoters and a number of tissue-specific transcripts have been identified. The gene is polymorphic with most mutations identified to date producing an unstable protein that is subject to polyubiquitination. The NAT1 protein contains a catalytic triad similar to a number of cysteine proteases and transglutaminases. NAT1 is widely distributed in the body, but the only endogenous substrate identified to date is the folate catabolite p-aminobenzoylglutamate. Recent links between NAT1 genotypes and susceptibility to spina bifida suggests that the enzyme has an important role in folate homeostasis. (C) 2007 Elsevier Ltd. All fights reserved. 2008-02-18T15:23:25Z Minchin, RF; Hanna, PE; Dupret, JM; Wagner, CR; Rodrigues-Lima, F; Butcher, NJ Arylamine N-Acetyltransferases http://espace.library.uq.edu.au/view/UQ:195908 2010-02-19T11:53:43Z Butcher, Neville J.; Minchin, Rodney F. A seasonal profile of plasma triglyceride levels in nesting flatback (Natator depressus) turtles on Curtis Island, Queensland, Australia http://espace.library.uq.edu.au/view/UQ:148545 2008-06-06T14:37:19Z Slater, L.; Limpus, C.; Whittier, J. M. A serial analysis of gene expression profile of the Alzheimer's disease Tg2576 mouse model http://espace.library.uq.edu.au/view/UQ:201449 Serial analysis of gene expression (SAGE), a technique that allows for the simultaneous detection of expression levels of the entire genome without a priori knowledge of gene sequences, was used to examine the transcriptional expression pattern of the Tg2576 mouse model of Alzheimer’s disease (AD). Pairwise comparison between the Tg2576 and nontransgenic SAGE libraries identified a number of differentially expressed genes in the Tg2576 SAGE library, some of which were not previously revealed by the microarray studies. Real-time PCR was used to validate a panel of genes selected from the SAGE analysis in the Tg2576 mouse brain, as well as the hippocampus and temporal cortex of sporadic AD and normal age-matched controls. NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 5 (NDUFA5) and FXYD domain-containing ion transport regulator 6 (FXYD6) were found to be significantly decreased in the Tg2576 mouse brain and AD hippocampus. PTEN-induced putative kinase 1 (PINK1), phosphatidylethanolamine binding protein (PEBP), crystalline μ (CRYM), and neurogranin (NRGN) were significantly decreased in AD tissues. The gene ontologies represented in the Tg2576 data were statistically analyzed and demonstrated a significant under-representation of genes involved with G-protein-coupled receptor signaling and odorant binding, while genes significantly over-represented were focused on cellular communication and cellular physiological processes. The novel approach of profiling the Tg2576 mouse brain using SAGE has identified different genes that could subsequently be examined for their potential as peripheral diagnostic and prognostic markers for Alzheimer’s disease. 2010-03-31T16:12:32Z George, Amee; Gordon, Lavinia; Beissbarth, Tim; Koukoulas, Irene; Holsinger, R.; Perreau, Victoria; Cappai, Roberto; Tan, Seong-Seng; Masters, Colin; Scott, Hamish; Li, Qiao-Xin A set of vertically integrated inquiry-based practical curricula that develop scientific thinking skills for large cohorts of undergraduate students http://espace.library.uq.edu.au/view/UQ:319741 Science graduates require critical thinking skills to deal with the complex problems they will face in their 21st century workplaces. Inquiry-based curricula can provide students with the opportunities to develop such critical thinking skills; however, evidence suggests that an inappropriate level of autonomy provided to underprepared students may not only be daunting to students but also detrimental to their learning. After a major review of the Bachelor of Science, we developed, implemented, and evaluated a series of three vertically integrated courses with inquiry-style laboratory practicals for early-stage undergraduate students in biomedical science. These practical curricula were designed so that students would work with increasing autonomy and ownership of their research projects to develop increasingly advanced scientific thinking and communication skills. Students undertaking the first iteration of these three vertically integrated courses reported learning gains in course content as well as skills in scientific writing, hypothesis construction, experimental design, data analysis, and interpreting results. Students also demonstrated increasing skills in both hypothesis formulation and communication of findings as a result of participating in the inquiry-based curricula and completing the associated practical assessment tasks. Here, we report the specific aspects of the curricula that students reported as having the greatest impact on their learning and the particular elements of hypothesis formulation and communication of findings that were more challenging for students to master. These findings provide important implications for science educators concerned with designing curricula to promote scientific thinking and communication skills alongside content acquisition. 2013-12-18T14:24:58Z Zimbardi, Kirsten; Bugarcic, Andrea; Colthorpe, Kay; Good, Jonathon P; Lluka, Lesley A short attention span in the memory consolidation mutant radish http://espace.library.uq.edu.au/view/UQ:216483 2010-09-16T15:07:03Z van Swinderen, B.; Brembs, B. A shuffled CYP1A library shows both structural integrity and functional diversity http://espace.library.uq.edu.au/view/UQ:130674 The cytochrome P450 enzymes (P450s) that mediate mammalian xenobiotic metabolism are highly versatile monooxygenases, which show wide and overlapping substrate ranges but generally poor catalytic rates. Re-engineering of these P450s may enable the development of useful biocatalysts for industrial applications. In the current study, restriction enzyme-mediated DNA family shuffling was used to create a library from human CYP1A1 and CYP1A2. Among sequenced clones (four randomly selected and eight functional clones), 5.9 +/- 2.3 crossovers and 1.5 +/- 1.5 spontaneous mutations (mean +/- S.D.) were detected per mutant. A high level of structural integrity as well as diverse functionality were found, with 53% of clones expressed at significant levels (> 50 nM P450 hemoprotein) and 23% of clones showing activity on one or more of the following compounds: luciferin 6'-chloroethyl ether (luciferin-CEE), luciferin 6'-methyl ether (luciferin-ME), 6'-deoxyluciferin (luciferin-H), the ethylene glycol ester of luciferin 6'-methyl ether, 7-ethoxyresorufin, and p-nitrophenol (PNP). Different activity profiles were seen with higher specific activity on individual compounds (e.g., clone 22; 9 times the CYP1A1 specific activity toward luciferin-CEE), novel activities (e.g., clone 35; activity toward luciferin-H and PNP), and broadening of substrate range observed in particular clones (e.g., clone 9; activity toward both selective substrates luciferin-ME and luciferin-CEE as well as toward luciferin-H and PNP). In summary, forms were found with distinct and novel activity profiles, despite the relatively small number of mutants examined. In addition, the whole-cell metabolic assays described here provide simple, high-throughput methods useful for screening larger libraries. 2008-02-18T15:42:33Z Johnston, Wayne A.; Huang, Weiliang; De Voss, James J.; Hayes, Martin A.; Gillam, Elizabeth M.J. A shuffled CYP2C library with a high degree of structural integrity and functional versatility http://espace.library.uq.edu.au/view/UQ:130669 Cytochrome P450 (CYP) enzymes involved in mammalian xenobiotic metabolism are attractive targets for the engineering of biocatalysts since they have broad and overlapping substrate and reaction substrate specificities. In this report, a library of chimeric mutants was prepared from CYP2C8, CYP2C9, CYP2C18 and CYP2C19 by DNA family shuffling. Twelve randomly selected clones were fully sequenced and showed 9 +/- 2 crossovers and 1.5 +/- 0.5 spontaneous mutations per similar to 1.5 kbp open reading frame. CYP hemoprotein expression was observed in 50% (microaerobic culture) to 54% (aerobic culture) of clones. The functional diversity of the library was assessed using three luminogenic substrates, diclofenac and indole as probe substrates. A random sample of 26 clones revealed two clones with activity towards luciferin ME, one towards luciferin H and five towards diclofenac 4'-hydroxylation. One mutant showed activity towards all three substrates. Of 96 clones screened on solid media, one showed elevated indigo production compared to the parental forms. Turnover rates for luciferin ME and H metabolism by CYP2C9 and mutants were at least one order of magnitude higher in experiments with membranes compared to whole cells, consistent with impaired product egress from cells. Apparent K-m values were increased in whole cell incubations with luciferin H suggesting impaired access of the substrate to the active site of the enzymes in whole cells. Finally screening with a panel of CYP2C ligands using CYP2C9 or active mutants revealed different patterns of inhibition and heteroactivation of metabolism of luciferin analogs. (C) 2007 Elsevier Inc. All rights reserved. 2008-02-18T15:42:12Z Huang, Weiliang; Johnston, Wayne A.; Hayes, Martin A.; De Voss, James J.; Gillam, Elizabeth M.J. A simple and reliable electroporation method for human bone marrow mesenchymal stem cells http://espace.library.uq.edu.au/view/UQ:177069 2009-04-20T04:18:13Z Helledie, T.; Nurcombe, V.; Cool, S. M. A simple approach to prepare monodisperse mesoporous silica nanospheres with adjustable sizes http://espace.library.uq.edu.au/view/UQ:274804 2012-05-28T23:19:22Z Yu, Meihua; Zhou, Liang; Zhang, Jun; Yuan, Pei; Thorn, Peter; Gu, Wenyi; Yu, Chengzhong A simple magnetic separation method for high-yield isolation of pure primary microglia http://espace.library.uq.edu.au/view/UQ:329033 2014-04-30T23:39:49Z Gordon, Richard; Hogan, Colleen E.; Neal, Matthew L.; Anantharam, Vellareddy; Kanthasamy, Anumantha G.; Kanthasamy, Arthi A simple way to cultivate referencing habits in first year biology students http://espace.library.uq.edu.au/view/UQ:345759 2014-11-25T05:06:03Z Chunduri, Prasad; Lluka, Lesley; Kinna, Genevieve; Good, Jonathan; Zimbardi, Kirsten; Colthorpe, Kay A single amino acid substitution in the hemagglutinin of H3N2 subtype influenza A viruses is associated with resistance to the long pentraxin PTX3 and enhanced virulence in mice http://espace.library.uq.edu.au/view/UQ:348163 2014-12-30T23:19:09Z Job, Emma R.; Bottazzi, Barbara; Short, Kirsty R.; Deng, Yi-Mo; Mantovani, Alberto; Brooks, Andrew G.; Reading, Patrick C. A single beta-amino acid substitution to angiotensin II confers AT(2) receptor selectivity and vascular function http://espace.library.uq.edu.au/view/UQ:256740 Novel AT2R ligands were designed by substituting individual β-amino acid in the sequence of the native ligand angiotensin II (Ang II). Relative ATR selectivity and functional vascular assays (in vitro AT2R-mediated vasorelaxation and in vivo vasodepressor action) were determined. In competition binding experiments using either AT1R- or AT2R- transfected HEK-293 cells, only β-Asp-Ang II and Ang II fully displaced [I]-Ang II from AT1R. In contrast, β-substitutions at each position of Ang II exhibited AT2R affinity, with β-Tyr-Ang II and β-Ile-Ang II exhibiting 1000-fold AT2R selectivity. In mouse aortic rings, β-Tyr-Ang II and β-Ile-Ang II evoked vasorelaxation that was sensitive to blockade by the AT2R antagonist PD123319 and the nitric oxide synthase inhibitor L-NAME. When tested with a low level of AT1R blockade, β-Ile-Ang II (15 pmol/kg per minute IV for 4 hours) reduced blood pressure (BP) in conscious spontaneously hypertensive rats (β-Ile-Ang II plus candesartan, -24±4 mm Hg) to a greater extent than candesartan alone (-11±3 mm Hg, n=7, P<0.05), an effect that was abolished by concomitant PD123319 infusion. However, in an identical experimental protocol, β-Tyr-Ang II had no influence on BP (n=10), and it was less stable than β-Ile-Ang II in plasma stability assays. Thus, this study demonstrated that a single β-amino acid substitution resulted in a compound that demonstrated both in vitro vasorelaxation and in vivo depressor activity via AT2R. This approach to the design and synthesis of novel AT2R-selective peptidomimetics shows great potential to provide insight into AT2R function. 2011-10-18T13:30:38Z Jones, Emma S.; Del Borgo, Mark P.; Kirsch, Julian F.; Clayton, Daniel; Bosnyak, Sanja; Welungoda, Iresha; Hausler, Nicholas; Unabia, Sharon; Perlmutter, Patrick; Thomas, Walter G.; Aguilar, Marie-Isabel; Widdop, Robert E. A single beta subunit M2 domain residue controls the picrotoxin sensitivity of alpha beta heteromeric glycine receptor chloride channels http://espace.library.uq.edu.au/view/UQ:60872 This study investigated the residues responsible for the reduced picrotoxin sensitivity of the alpha beta heteromeric glycine receptor relative to the alpha homomeric receptor. By analogy with structurally related receptors, the beta subunit M2 domain residues P278 and F282 were considered the most likely candidates for mediating this effect. These residues align with G254 and T258 of the alpha subunit. The T258A, T258C and T258F mutations dramatically reduced the picrotoxin sensitivity of the alpha homomeric receptor. Furthermore, the converse F282T mutation in the beta subunit increased the picrotoxin sensitivity of the alpha beta heteromeric receptor. The P278G mutation in the beta subunit did not affect the picrotoxin sensitivity of the alpha beta heteromer. Thus, a ring of five threonines at the M2 domain depth corresponding to alpha subunit T258 is specifically required for picrotoxin sensitivity. Mutations to alpha subunit T258 also profoundly influenced the apparent glycine affinity. A substituted cysteine accessibility analysis revealed that the T258C sidechain increases its pore exposure in the channel open state. This provides further evidence for an allosteric mechanism of picrotoxin inhibition, but renders it unlikely that picrotoxin las an allosterically acting 'competitive' antagonist) binds to this residue. 2007-08-14T16:50:12Z Shan, Q.; Haddrill, J. L.; Lynch, J. W. A single-chain derivative of the relaxin hormone is a functionally selective agonist of the G protein-coupled receptor, RXFP1 http://espace.library.uq.edu.au/view/UQ:393291 2016-07-03T00:19:17Z Hossain, Mohammed Akhter; Kocan, Martina; Yao, Song T.; Royce, Simon G.; Nair, Vinojini B.; Siwek, Christopher; Patil, Nitin A.; Harrison, Ian P.; Rosengren, K. Johan; Selemidis, Stavros; Summers, Roger J.; Wade, John D.; Bathgate, Ross A. D.; Samuel, Chrishan S. A small molecule angiotensin II type 2 receptor (AT2R) antagonist produces analgesia in a rat model of neuropathic pain by inhibition of p38 Mitogen-Activated Protein Kinase (MAPK) and p44/p42 MAPK activation in the dorsal root ganglia http://espace.library.uq.edu.au/view/UQ:311863 2013-10-09T12:06:41Z Smith, Maree T.; Woodruff, Trent M.; Wyse, Bruce D.; Muralidharan, Arjun; Walther, Thomas A small molecule C5a receptor antagonist protects kidneys from ischemia/reperfusion injury in rats http://espace.library.uq.edu.au/view/UQ:67419 2007-08-15T02:41:30Z Arumugam, T. V.; Shiels, I. A.; Strachan, A. J.; Abbenante, G.; Fairlie, D. P.; Taylor, S. M. A species of reef fish that uses ultraviolet patterns for covert face recognition http://espace.library.uq.edu.au/view/UQ:200943 The evolutionary and behavioral significance of an animal's color patterns remains poorly understood [1-4], not least, patterns that reflect ultraviolet (UV) light [5]. The current belief is that UV signals must be broad and bold to be detected because (1) they are prone to scattering in air and water, (2) when present, UV-sensitive cones are generally found in low numbers, and (3) long-wavelength-sensitive cones predominate in form vision in those species tested to date [6]. We report a study of two species of damselfish whose appearance differs only in the fine detail of UV-reflective facial patterns. We show that, contrary to expectations, the Ambon damselfish (Pomacentrus amboinensis) is able to use these patterns for species discrimination. We also reveal that the essential features of the patterns are contained in their shape rather than color. The results provide support for the hypothesis that UV is used by some fish as a high-fidelity "secret communication channel" hidden from predators [7, 8]. In more general terms, the findings help unravel the details of a language of color and pattern long since lost to our primate forebears, but which has been part of the world of many seeing organisms for millions of years. 2010-03-28T00:06:18Z Siebeck, Ulrike E.; Parker, Amira N.; Sprenger, Dennis; Mathger, Lydia M.; Wallis, Guy Aspects of reproduction and diet of the Australian endemic skate Dipturus polyommata (Ogilby) (Elasmobranchii : Rajidae), by-catch of a commercial prawn trawl fishery http://espace.library.uq.edu.au/view/UQ:152109 2008-07-29T16:19:37Z Kyne, P. M.; Courtney, A. J.; Bennett, M. B. Aspects of the life history of epqulette sharks, Hemiscyllium ocellatum, on Heron Island Reef, Great Barrier Reef, Australia http://espace.library.uq.edu.au/view/UQ:150489 2008-06-06T16:33:56Z Heupel, M.; Bennett, M. B. | {
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-5,852,583,869,018,107,000 | CALL CENTRE: +27(0)861111866
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Wolwespruit Nature Reserve
Click to download the brochureContact Information:
Tel: +27(0)18 581 9705
Email: [email protected] / [email protected]
Postal Address:
PO Box 237, Leeudoringstad, 2640
Address:
Leeudoringstad turn-off, Klipspruit road.
Click to download the reserve brochure
This small but scenic reserve conserves attractive riverine bush and is a popular angling and hiking destination, along the tranquil river frontage environment.
Wolwespruit is one of the least known of the North West parks and has retained much of the appeal of the natural riverine environment. It also conserves a variety of plains game.
Wolwespruit Nature Reserve is well known for yellowfish angling in the rapids with artificial lures and flies.
.
Steenbuck or Phudufudu at Wolwespruit Nature Reserve
Wolwespruit Nature Reserve
Location
Situated on the Vaal River upstream 130km from the Bloemhof Dam and 22 km south from the town of Leeudoringstad, on the Klipspruit road.
Climate
Generally mild to hot but can be cold in winter especially at night.
The birding opportunities are numerous and diverse in the various localised eco-systems. The park also conserves a variety of plains game and smaller mammals.
Facilities
There are rustic camping sites all along the river bank on the reserve and a self catering stone cottage sleeping 6 people. Booking essential.
Mafikeng
19°
clear sky
humidity: 28%
wind: 2m/s N
H 32 • L 21
23°
Wed
20°
Thu
22°
Fri
21°
Sat
Weather from OpenWeatherMap
...powerD by 3rdEFX
Translate » | {
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-5,588,104,010,201,519,000 | The Facts on File Dictionary of Biology
The Facts on File Dictionary of Biology
Book - 1999
Rate this:
Baker & Taylor
A reference tool for professional and student biologists spans everything from abaxial to zymogen granules, explaining such biological phenomena as photosynthesis, regeneration, and cytoplasmic inheritance
Book News
The third edition of the dictionary now contains 3300 headwords covering the terminology of modern biology, and features an appendix with charts of the animal kingdom, the plant kingdom, and amino-acid structures. Annotation c. Book News, Inc., Portland, OR (booknews.com)
Publisher: New York : Facts On File, 1999
Edition: 3rd ed
ISBN: 9780816039081
0816039089
9780816039074
0816039070
Branch Call Number: R 570.3 FAC
Characteristics: 361 p. : ill. ; 24 cm
Additional Contributors: Hine, Robert
Facts on File, Inc
Alternative Title: Biology
Dictionary of biology
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7,265,488,639,130,329,000 | To view, register or book classes, please use a desktop or laptop device for best results
MS/HS Marine Biology
Teacher: Ms. Kay
Join us on a virtual tour of Earth’s ocean ecosystems! During this 13 week course, we’ll learn the basic physics, biology, and chemistry of our oceans. We’ll explore habitats that include the high energy, extreme intertidal zone, deep sea hydrothermal vents, polar seas, kelp forests and coral reefs. Life evolved in the sea, and our oceans are home to many amazing groups of animals that are never seen on land. This class will survey the incredible range of phyla found there – from single-celled dinoflagellates (that can be both photosynthetic and predatory – and make their own light) to the largest mammal on earth, the blue whale. We’ll also learn the integral role earth’s oceans play in regulating climate, and how climate change is affecting these ecosystems.
Join us for some challenging and fun hands-on science! This class is designed as a survey course, to give the students a broad overview of the subject, so we’ll talk about physics concepts, but we won’t get into the specific mathematical equations involved. If your student is not a physics/math fan, they’ll be perfectly happy with the level we’re approaching things at.
Class is live online and labs will be held outside with social distancing and masks once a month in Springfield, VA. The option to do the in-person labs virtually is available.
Material fee: $10 supply fee
Instructor: The Science Place, Kaira Harris
Minimum enrollment requirement: 4
Course Length: 50 mins per week /16 weeks / semester
Cost: $208+ $10 supply fee = $218 per semester
SKU: N/A Category:
$218.00
Course Booking
Clear
Select Students for Booking
To purchase a class you need to have an account with a student added. Add a student or register. | {
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941,594,963,313,203,000 | Anti-Hu CD56 Pacific Blue™
Anti-Hu CD56 Pacific Blue™
Regulatory status
RUO
Antigen
CD56
Clone
LT56
Format
Pacific Blue™
Reactivity
Human
Application
Excitation laser
violet (405 nm)
Variant
100 tests
PB-789-T100
In stock
220.00 USD
25 tests
PB-789-T025
Delivery 1 week
110.00 USD
Variant
100 tests
PB-789-T100
In stock
220.00 USD
25 tests
PB-789-T025
Delivery 1 week
110.00 USD
Product details
Description
Images
References
SDS download
Isotype
Mouse IgG2a
Specificity
The mouse monoclonal antibody LT56 recognizes an extracellular epitope of CD56 (NCAM), a transmembrane glycoprotein expressed ubiquitously in the nervous system and found also on T cells and NK cells.
Workshop
HLDA X
Application
Application details
Flow cytometry: The reagent is designed for analysis of human blood cells using 4 μl reagent / 100 μl of whole blood or 106 cells in a suspension. The content of a vial (0.4 ml) is sufficient for 100 tests.
Reactivity
Human
Immunogen
Cell line KG1a
Preparation
Purified antibody is conjugated with Pacific Blue™ NHS ester under optimum conditions and unconjugated antibody and free fluorochrome are removed by size-exclusion chromatography.
Formulation
Stabilizing phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Storage and handling
Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.
Exbio licence note
The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.
Licence note
Alexa Fluor®, Pacific Blue™ and Pacific Orange™ are registered trademarks of Life Technologies Corporation.
Licence label
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.
Other names
NCAM1, MSK39
Antigen description
CD56 (NCAM, neural cell adhesion molecule) is a transmembrane glycoprotein of immunoglobulin family serving as adhesive molecule which is ubiquitously expressed in nervous system, usually as 120 kDa, 140 kDa or 180 kDa isoform, and it is also found on T cells and NK cells. Polysialic modification results in reduction of CD56-mediated cell adhesion and is involved in cell migration, axonal growth, pathfinding and synaptic plasticity. CD56 is a widely used neuroendocrine marker with a high sensitivity for neuroendocrine tumours and ovarian granulosa cell tumours.
Entrez Gene ID 4684
UniProt ID P13591
PB-789 FC
Flow cytometry analysis (surface staining) of human peripheral blood with anti-human CD56 (LT56) Pacific Blue™.
General references:
McCluggage WG, McKenna M, McBride HA: CD56 is a sensitive and diagnostically useful immunohistochemical marker of ovarian sex cord-stromal tumors. Int J Gynecol Pathol. 2007 Jul;26(3):322-7.
PubMed
Ohishi Y, Kaku T, Oya M, Kobayashi H, Wake N, Tsuneyoshi M: CD56 expression in ovarian granulosa cell tumors, and its diagnostic utility and pitfalls. Gynecol Oncol. 2007 Oct;107(1):30-8.
PubMed
Jakovcevski I, Mo Z, Zecevic N: Down-regulation of the axonal polysialic acid-neural cell adhesion molecule expression coincides with the onset of myelination in the human fetal forebrain. Neuroscience. 2007 Oct 26;149(2):328-37.
PubMed
Lanier LL, Chang C, Azuma M, Ruitenberg JJ, Hemperly JJ, Phillips JH: Molecular and functional analysis of human natural killer cell-associated neural cell adhesion molecule (N-CAM/CD56). J Immunol. 1991 Jun 15;146(12):4421-6.
PubMed
Product specific references:
Munteanu AN, Surcel M, Huică RI, Isvoranu G, Constantin C, Pîrvu IR, Chifiriuc C, Ulmeanu C, Ursaciuc C, Neagu M: Peripheral immune cell markers in children with recurrent respiratory infections in the absence of primary immunodeficiency. Exp Ther Med 2019, June 26, 1693-1700
PubMed
Variant
100 tests
PB-789-T100
In stock
220.00 USD
25 tests
PB-789-T025
Delivery 1 week
110.00 USD
Variant
100 tests
PB-789-T100
In stock
220.00 USD
25 tests
PB-789-T025
Delivery 1 week
110.00 USD
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190.00 USD | {
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-6,858,558,568,201,226,000 | Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You and Your Team.
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FEMS Microbiology Reviews Volume 24 Issue 3
Subject Areas: Microbiology; Medicine; Infectious Diseases
Publisher: Blackwell - Oxford University Press
ISSN: 0168-6445
1 - 5 of 5 articles
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4,027,626,612,822,714,000 | The Scientist
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-3,223,165,443,934,651,400 | Search form
Fall colors on Mirror Lake
How Much Oxygen Is in Our Lakes and Streams?
If you spend time following the work of the Ausable River Association, you know that dissolved oxygen (DO) is an important component of water quality. Dissolved oxygen is the measure of how much oxygen is dissolved in water, or the amount of oxygen that is available to aquatic organisms. Oxygen is vital to almost all forms of life including aquatic insects, zooplankton, and fish. In general, dissolved oxygen levels are high in rapidly moving water and low in still or stagnant waters. Precision methods exist for measuring DO in water and can be compared to tested standards for supporting specific species in ideal and in stressed environments.
Dissolved oxygen is measured with a field or lab meter and expressed as a concentration, specifically in mg/L or as a percent of saturation. DO concentrations typically range from 0 mg/L to 25 mg/L. Aquatic organisms such as lake trout prefer DO concentrations greater than 6 mg/L. Less than 2 mg/L of DO are considered anoxic or low oxygen and can be lethal to organisms.
Dissolved oxygen levels in rivers such as the Ausable are typically high due to the turbulent water flow. However, increases in soluble elements, excessive bacterial growth due to elevated nutrient inputs, or higher water temperatures, can cause decreases in DO. The solubility of oxygen in water is directly related to temperature and salinity and decreases as the others increase.
Graph of temperature and dissolved oxygen from April 2015- August 2021 in Styles Brook.
The graph above shows DO concentrations (orange line) and water temperature (blue line) in Styles Brook on the East Branch Ausable River in the towns of Keene and Jay from April 2015 to August 2021. The direct relationship between dissolved DO and temperature can be seen as the lines alternate in direction. Additionally, DO and temperature vary by season and year. Higher DO concentrations correspond to lower temperatures in the fall and winter. One of the warmest years on record was in 2018, which correlates with lower DO concentrations. Preliminary data for late 2020 and early 2021 show higher DO levels, we are working to understand this rise. But, it is likely due to snow melt and turbulent water from the high gradient of Styles Brook. Dissolved oxygen levels are important to track as lower DO leads to stressed aquatic species which can slow metabolic rates and reproduction.
Support our water quality work for clean water and healthy streams. Give with confidence today!
As in streams, primary sources of DO in lakes occur through diffusion from the atmosphere and primary production. These processes vary throughout the year. During periods of lake turnover in the spring and fall, oxygen is redistributed throughout the water column. During the summer stratified period, warm water at the surface of the lake prevents the cold bottom waters from mixing and limits the source of atmospheric oxygen. Similarly, during the winter months, while the lake is covered with ice, the supply of atmospheric oxygen is reduced and contributions from primary production are very low.
Temperature (°C) (A) and dissolved oxygen (mg/L) (B) from vertical profiles collected at 1m intervals at the deepest part of Mirror lake for 2020. Lighter colors denotes higher temperatures or dissolved oxygen.Courtesy of Brendan Wiltse.
In Mirror Lake, the subject of the above graph, reductions in bottom water DO raise concerns for aquatic wildlife, especially native salmonid species. During the summer months, these fish species seek out an optimal zone in the lake, an area which is both cool (<15 °C) and well oxygenated (>6 mg/L). This zone can be very small, or non-existent in lakes. There are parallels for native fish and other aquatic species in streams. Our work at the Ausable River Association uses rigorous science to understand human and climate induced changes in waterways and, working with our communities, identifies innovative solutions that can lead to healthier habitats for native aquatic species.
Story, photos and first graph by Leanna Thalmann, Water Quality Associate. Top photo: Fall colors on Mirror Lake.
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} | 57 | ff2a0dd8e83e1035faba2b053f3eae93 |
4,379,646,983,460,181,500 | scientific teaching models
Friday, 18 December 2015 Wednesday, 08 December 2021
Visual scientific models inspired your time at school mainly in biology classes. Can you remember the feeling unpicking a model of an ear piece per piece? You and your friends could play ´being a doctor` on another level. These beautiful scientific models made of wood and plaster with their realistic looks supported teachers for decades to highlight their learning material. Rediscover the schoolkid in yourself and pause for our lovely picture gallery....
scientific teaching models
Anatomical models are three-dimensional, homolog models which depict the anatomical structure life-sized or enlarged. These models are not merely mainly visual aids but vital teaching tools for medical skills at universities and hospitals. Antique and or vintage scientific models however can be part of an individual furnishing and artfour luckily possesses some really exceptional pieces. Just take a look.
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-2,072,626,325,009,697,500 | Share this post on:
Cytes in response to interleukin-2 stimulation50 delivers however a different instance. four.2 Chemistry of DNA demethylation In contrast to the well-studied biology of DNA methylation in mammals, the enzymatic mechanism of active demethylation had extended remained elusive and controversial (reviewed in 44, 51). The fundamental chemical challenge for direct removal on the 5-methyl group from the pyrimidine ring is usually a higher stability in the C5 H3 bond in water beneath physiological conditions. To obtain around the unfavorable nature of your direct cleavage of the bond, a cascade of coupled reactions might be utilised. For example, certain DNA order BIP-V5 repair enzymes can reverse N-alkylation harm to DNA via a two-step mechanism, which entails an enzymatic oxidation of N-alkylated nucleobases (N3-alkylcytosine, N1-alkyladenine) to corresponding N-(1-hydroxyalkyl) derivatives (Fig. 4D). These intermediates then undergo spontaneous hydrolytic release of an aldehyde from the ring nitrogen to straight create the original unmodified base. Demethylation of biological methyl marks in histones occurs by way of a similar route (Fig. 4E) (reviewed in 52). This illustrates that oxygenation of theChem Soc Rev. Author manuscript; out there in PMC 2013 November 07.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptKriukien et al.Pagemethylated goods leads to a substantial weakening in the C-N bonds. However, it turns out that hydroxymethyl groups attached towards the 5-position of pyrimidine bases are however chemically stable and long-lived below physiological situations. From biological standpoint, the generated hmC presents a kind of cytosine in which the proper 5-methyl group is no longer present, but the exocyclic 5-substitutent just isn’t removed either. How is this chemically stable epigenetic state of cytosine resolved? Notably, hmC will not be recognized by methyl-CpG binding domain proteins (MBD), including the transcriptional repressor MeCP2, MBD1 and MBD221, 53 suggesting the possibility that conversion of 5mC to hmC is adequate for the reversal of the gene silencing effect of 5mC. Even within the presence of upkeep methylases for example Dnmt1, hmC would not be maintained after replication (passively removed) (Fig. 8)53, 54 and would be treated as “unmodified” cytosine (with a distinction that it cannot be directly re-methylated without having prior removal of the 5hydroxymethyl group). It is actually reasonable to assume that, although being produced from a major epigenetic mark (5mC), hmC may play its personal regulatory function as a secondary epigenetic mark in DNA (see examples below). Although this scenario is operational in certain cases, substantial evidence indicates that hmC could possibly be further processed in vivo to ultimately yield unmodified cytosine (active demethylation). It has been shown not too long ago that Tet proteins possess the capacity to further oxidize hmC forming fC and caC in vivo (Fig. 4B),13, 14 and modest quantities of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/21215484 these solutions are detectable in genomic DNA of mouse ES cells, embyoid bodies and zygotes.13, 14, 28, 45 Similarly, enzymatic removal in the 5-methyl group inside the so-called thymidine salvage pathway of fungi (Fig. 4C) is achieved by thymine-7-hydroxylase (T7H), which carries out 3 consecutive oxidation reactions to hydroxymethyl, and then formyl and carboxyl groups yielding 5-carboxyuracil (or iso-orotate). Iso-orotate is lastly processed by a decarboxylase to give uracil (reviewed in).44, 52 To date, no orthologous decarboxylase or deformylase activity has been.
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Author: DGAT inhibitor | {
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-1,555,140,457,652,972,800 | Team:Mingdao/Parts
In iGEM 2016, we designed and created a novel BLOOD ALCOHOL METER (iMeter) inspired by blood glucose meter (BGM). The glucose oxidase (GOX) was used as an oxidoreductase to catalyze the oxidation of glucose in the electrochemical analyzer. In a similar way, the alcohol oxidase (AOX) is an oxidoreductase to catalyze the oxidation of alcohol and could be applied in the alcohol measurements. This year, we proved the concept by cloning genes, protein analysis, enzyme activity assay and electrochemical modeling.
For the BioBrick parts, we’ve created and submitted 11 new parts to the Registry including AOX genes [BBa_K1991000, BBa_K1991001], Lpp-OmpA (LO) genes [BBa_K1991004] and LO-AOX [BBa_K1991005, BBa_K1991006] fusion protein genes as Basic Parts, as well as a constitutive promoter [BBa_J23101]and RBS [BBa_B0034]-driven LO fusion protein genes [BBa_K1991002, BBa_K1991003, BBa_K1991007, BBa_K1991008, BBa_K1991009, BBa_K1991010] as Composite Parts. We have demonstrated and collected all of these parts for the application of alcohol oxidase in the blood alcohol meter.
AOX1/pSB1C3
Registry: BBa_K1991000
Source: Pichia pastoris
Codon Optimization: E. coli codon preference
DNA synthesis: gBlocks® Gene Fragments (IDT)
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX1-XbaI-F: 5’- GGCGAGTCTAGATGGCTATTCCGGAAGAGTT -3’
2. AOX1-PstI-R: 5’- AAAATTCTGCAGCGGCCGCTACTAGTATCAAAAGCGCGCAA - 3’
Cloning Procedure:
The synthesized AOX1 gene was amplified by PCR and digested by XbaI and PstI, followed by cloning onto pSB1C3 which was cut by XbaI and PstI. The part has been confirmed by sequencing.
AOX2/pSB1C3
Registry: BBa_K1991001
Source: Pichia pastoris
Codon Optimization: E. coli codon preference
DNA synthesis: gBlocks® Gene Fragments (IDT)
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX2-XbaI-F: 5’- GGAGAGTCTAGATGGCCATCCCTGAGGAATTTG -3’
2. AOX2-PstI-R: 5’- AAAAAACTGCAGCGGCCGCTACTAGTATCAGAAGCGCGCTAA -3’
Cloning Procedure:
The synthesized AOX2 gene was amplified by PCR and digested by XbaI and PstI, followed by cloning onto pSB1C3 which was cut by XbaI and PstI. The part has been confirmed by sequencing.
Lpp-OmpA-BamHI/pSB1C3
Registry: BBa_K1991004
Source: Escherichia coli
Original Part: BBa_K1694035
Notebook: PDF; Sequencing: VF2
Primers:
1. Lpp-XbaI-EcoRI-F: 5’- AGTCAAGAATTCGCGGCCGCTTCTAGATGAAAGCAACCAAGCTG -3’
2. OmpA-BamHI-SpeI-R: 5’- AAAGAGACTAGTAGGATCCACGTGTGGCAATTTCCGGGGTGA -3’
Cloning Procedure:
The DNA fragment of Lpp-OmpA (LO) was amplified by PCR using BBa_K1694035 as a template with a primer containing BamHI site and digested by EcoRI and SpeI, followed by cloning onto pSB1C3 which was cut by EcoRI and SpeI. The part has been confirmed by sequencing.
Existing part from NCTU-Formosa in 2015:
Improved part by Mingdao in 2016:
Lpp and OmpA are outer membrane proteins of E. coli. Lpp-OmpA (LO) hybrid can direct heterologous proteins to bacterial cell surface. In 2015, NCTU-Formosa used it to display scFv (single chain fragment variable) antibodies on the surface of E. coli. They found that a fusion protein cannot be possible to be created under the standard BioBrick assembly rule, that is EcoRI(E)-XbaI(X)-GENE-SpeI(S)-PstI(P). The A part of EX-LO-SP and the B part of EX-scFV-SP, for example, are connected by cutting and ligation of SpeI plus PstI for the A part and XbaI plus PstI for the B part. The SCAR generated by XbaI/SpeI (ACTAGA) will form a stop codon just in front of the ATG start codon of the scFV protein of the B part. This situation has been officially mentioned by the BioBrick standard assembly.
Figure 5: Limitation of cloning a fusion protein by standard biobrick assembly
Therefore, in 2015, NCTU-Formosa created a novel part (BBa_K1694002) putting NcoI site between the LO part and SpeI site. However, when considering cloning, we found that an extra NcoI site is present on Cm resistance gene making it difficult be a vector for gene cloning.
In 2016, Mingdao improved the part by replacing NcoI site with BamHI site (BBa_K1991004). Also, we’ve confirmed and prove the function of LO directing a fusion protein to the cell surface with enzyme activity in our project.
Figure 6: Alternative methods of cloning a fusion protein on Biobrick parts designed and created by NCTU-FORMOSA in 2015 and MINGDAO in 2016
BBa_K1991004: Lpp-OmpA-BamHI/pSB1C3 (PDF; VF2) The DNA fragment of Lpp-OmpA was amplified by PCR using BBa_K1694035 as a template with a primer containing BamHI site and digested by EcoRI and SpeI, followed by cloning onto pSB1C3 which was cut by EcoRI and SpeI. The part has been confirmed by sequencing.
BBa_K1991007: Pcons-RBS-LO-BamHI/pSB1C3 (PDF; VF2) The DNA fragment of Pcons-RBS-Lpp-OmpA was amplified by PCR using BBa_K1694035 as a template with a primer containing BamHI site and digested by EcoRI and SpeI, followed by cloning onto pSB1C3 which was cut by EcoRI and SpeI. The part has been confirmed by sequencing.
* Contributions: we’ve added the info to the original part's main page (BBa_K1694002). “An alternative version with BamHI site: If you'd like to directly use this part as a vector/backbone for cloning a fusion protein, you should be noticed that an extra NcoI site is present on Cm resistance gene. BBa_K1991004 replaced NcoI site behind LO with BamHI site and provides an alternative choice for cloning a LO fusion protein”
LO-AOX1-His/pSB1C3
Registry: BBa_K1991005
Source: Escherichia coli & Pichia pastoris
Original Part: BBa_K1991000, BBa_K1991004
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX1-BamHI-F: 5’- GAACGAGGATCCATGGCTATTCCGGAAGAGTT -3’
2. T7T-SpeI-PstI-R: 5’- AAGAGACTGCAGCGGCCGCTACTAGTATATAGTTCCTCCTTTCAG -3’
Cloning Procedure:
The DNA fragment of AOX1-His was amplified by PCR and digested by BamHI and PstI, followed by cloning onto LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing.
LO-AOX2-His/pSB1C3
Registry:BBa_K1991006
Source: Escherichia coli & Pichia pastoris
Original Part: BBa_K1991001, BBa_K1991004
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX2-BamHI-F: 5’- AAACGAGGATCCATGGCCATCCCTGAGGAATTTG -3’
2. T7T-SpeI-PstI-R: 5’- AAGAGACTGCAGCGGCCGCTACTAGTATATAGTTCCTCCTTTCAG -3’
Cloning Procedure:
The DNA fragment of AOX2-His was amplified by PCR and digested by BamHI and PstI, followed by cloning onto LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing.
Pcons-RBS-AOX1/pSB1C3
Registry: BBa_K1991002
Source: Pichia pastoris
Original Part: BBa_J23101, BBa_B0034, BBa_K1991000
Notebook: PDF; Sequencing: VF2
Primers:
1. Pcons-XbaI-EcoRI-F: 5’- AGTACAGAATTCGCGGCCGCTTCTAGAGTTTACAGCTAGCTCAG - 3’
2. RBS-SpeI-R: 5’- GGCGGCACTAGTATTTCTCCTCTTTCTCTAGTA -3’
Cloning Procedure:
The DNA fragment containing a constitutive promoter (BBa_J23101) and RBS (BBa_B0034) was amplified by PCR from BBa_K1694035 and digested by EcoRI and SpeI, followed by cloning onto AOX1/pSB1C3 which was cut by EcoRI and XbaI. The part has been confirmed by sequencing.
Pcons-RBS-AOX2/pSB1C3
Registry: BBa_K1991003
Source: Pichia pastoris
Original Part: BBa_J23101, BBa_B0034, BBa_K1991001
Notebook: PDF; Sequencing: VF2
Primers:
1. Pcons-XbaI-EcoRI-F: 5’- AGTACAGAATTCGCGGCCGCTTCTAGAGTTTACAGCTAGCTCAG - 3’
2. RBS-SpeI-R: 5’- GGCGGCACTAGTATTTCTCCTCTTTCTCTAGTA -3’
Cloning Procedure:
The DNA fragment containing a constitutive promoter (BBa_J23101) and RBS (BBa_B0034) was amplified by PCR from BBa_K1694035 and digested by EcoRI and SpeI, followed by cloning onto AOX2/pSB1C3 which was cut by EcoRI and XbaI. The part has been confirmed by sequencing.
Pcons-RBS-LO-BamHI/pSB1C3
Registry: BBa_K1991007
Source: Escherichia coli
Original Part: BBa_J23101, BBa_B0034, BBa_K1694035
Notebook: PDF; Sequencing: VF2
Primers:
1. Pcons-XbaI-EcoRI-F: 5’- AGTACAGAATTCGCGGCCGCTTCTAGAGTTTACAGCTAGCTCAG - 3’
2. OmpA-BamHI-SpeI-R: 5’- AAAGAGACTAGTAGGATCCACGTGTGGCAATTTCCGGGGTGA -3’
Cloning Procedure:
The DNA fragment of Pcons-RBS-Lpp-OmpA was amplified by PCR using BBa_K1694035 as a template with a primer containing BamHI site and digested by EcoRI and SpeI, followed by cloning onto pSB1C3 which was cut by EcoRI and SpeI. The part has been confirmed by sequencing.
Pcons-RBS-LO-AOX1-His/pSB1C3
Registry: BBa_K1991008
Source: Escherichia coli & Pichia pastoris
Original Part: BBa_J23101, BBa_B0034, BBa_K1991005
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX1-BamHI-F: 5’- GAACGAGGATCCATGGCTATTCCGGAAGAGTT -3’
2. T7T-SpeI-PstI-R: 5’- AAGAGACTGCAGCGGCCGCTACTAGTATATAGTTCCTCCTTTCAG -3’
Cloning Procedure:
The DNA fragment of AOX1-His was amplified by PCR and digested by BamHI and PstI, followed by cloning onto Pcons-RBS-LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing.
Pcons-RBS-LO-AOX2-His/pSB1C3
Registry: BBa_K1991009
Source: Escherichia coli & Pichia pastoris
Original Part: BBa_J23101, BBa_B0034, BBa_K1991006
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. AOX2-BamHI-F: 5’- AAACGAGGATCCATGGCCATCCCTGAGGAATTTG -3’
2. T7T-SpeI-PstI-R: 5’- AAGAGACTGCAGCGGCCGCTACTAGTATATAGTTCCTCCTTTCAG -3’
Cloning Procedure:
The DNA fragment of AOX2-His was amplified by PCR and digested by BamHI and PstI, followed by cloning onto Pcons-RBS-LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing.
To analyze the AOX gene expression, we run on a SDS-PAGE gel and observed by Coomassie Blue staining. The overnight-cultured E. coli were centrifuged and lysed with Lysis Buffer (12.5 mM Tris pH 6.8, 4% SDS). The resulting lysates were subjected to SDS-PAGE with a 10% polyacrylamide gel. The gel was stained with 0.25% Coomassie Brilliant Blue R250 for 2 hours and destained until the protein bands were clear. As the data showed in Figure 2, LO protein was expressed at around the estimated molecular weight of 17 kDa, LO-AOX fusion protein at 91 kDa and AOX protein at 79 kDa. The protein expression level was consistent with the data of GFP in Figure 1, demonstrating the low gene expression level of a LO fusion protein. However, so far we cannot confirm whether LO fusion protein is able to direct AOX or GFP proteins displayed on the cell surface of E. coli. We’re planning to do a subcellular fractionation to separate the outer membrane proteins for analysis in the future.
Figure 2: AOX protein analysis. SDS-PAG and Coomassie Blue staining were used to observe protein expression level. Lane 1: wild-type E. coli as a mock control; Lane 2: LO outer membrane protein [BBa_K1991007] (17 kDa) expression in E. coli; Lane 3: LO-AOX fusion protein [BBa_K1991009] (91 kDa) expression in E. coli.; Lane 4: AOX protein [BBa_K1991003] (79 kDa) expression in E. coli.
After AOX gene expression was confirmed, we want to know its enzyme activity. AOX catalyzes the oxidation of alcohol in the following chemical reaction C2H5OH + O2 CH3CHO + H2O2 and generates hydrogen peroxide. We examined the activity of AOX enzyme by measuring H2O2 production from the oxidation of ethanol. Fluorimetric Hydrogen Peroxide Assay Kit of Sigma-Aldrich was used for the study. In the kit, the red peroxidase substrate is designed to react with hydrogen peroxide to generate red fluorescence signal that can be detected at Ex/Em = 540/590 nm in the microplate reader. We prepared 1ml of overnight cultured E. coli displaying LO-AOX enzyme [BBa_K1991009]. The bacteria were centrifuged and resolved in Assay Buffer provided by the Sigma-Aldrich kit. Then, the resulting lysates were mixed with the increasing concentrations of ethanol from 0%, 0.025%, 0.05%, 0.075%, 0.10%, 0.15% to 0.2% for 3 minutes at room temperature. The tested alcohol concentrations were at the range from 0.03% (the alcohol law limit for safety driving) to 0.2% (people may lose consciousness at this level). The following procedure was according to the manufacture’s instruction. The data were read out at Ex/Em = 540/590 nm in the microplate reader (BioTek Microplate Spectrophotometer). As Figure 2 showed, the intensities of AOX activity were significantly and linearly correlated to the concentration of ethanol in a dose-dependent manner. The results indicated that LO-AOX enzyme we produced has a functional enzyme activity and reacted with alcohol concentration dose-dependently, implying we can use this assay to measure the unknown sample of alcohol.
Figure 2: AOX protein analysis. SDS-PAG and Coomassie Blue staining were used to observe protein expression level. Lane 1: wild-type E. coli as a mock control; Lane 2: LO outer membrane protein [BBa_K1991007] (17 kDa) expression in E. coli; Lane 3: LO-AOX fusion protein [BBa_K1991009] (91 kDa) expression in E. coli.; Lane 4: AOX protein [BBa_K1991003] (79 kDa) expression in E. coli.
The following is the message from iGEM team NCTU-FORMOSA:
“To test the enzyme activity of AOX protein, which catalyzes the oxidation of alcohol and generates H2O2 products, produced by iGEM Team Mingdao, we performed HRP-TMB assay. TMB (3,3',5,5'-Tetramethylbenzidine) is oxidized and changed to a deep blue color during the enzymatic degradation of H2O2 by horse radish peroxidase (HRP). The concentration of H2O2 can be determined by checking the color change, indicating TMB assay can be used to examine the enzyme activity of AOX with the substrate of alcohol.
We cultivated 3ml of the E.coli DH5α carrying the Mingdao iGEM team’s biobrick (i.e., Pcons-RBS-LO-AOX/pSB1C3, BBa_K1991009) in the LB media supplemented with Chloramphenicol. And we prepared another one as the control which didn’t have any plasmid. The next day, we centrifuged 1mL of bacteria expressing LO-AOX enzyme followed by being suspended in PBS. The control group was adjusted to the OD values at the same level of the LO-AOX group. After mixing bacterial samples with ethanol, TMB was added as a substrate of HRP and incubated in the dark for 3 min. The color of solution was significantly changed to blue at an OD650 of 1.51 compared to 1.21 in the control group, clearly demonstrating the ethanol was oxidized by AOX. As a result, we proved the function of their BioBrick by testing the AOX enzyme activity.”
Pcons-RBS-LO-GFP-His/pSB1C3
Registry: BBa_K1991010
Source: Escherichia coli & Aequorea victoria
Original Part: BBa_J23101, BBa_B0034, BBa_K1694035
Notebook: PDF; Sequencing: VF2, VR
Primers:
1. GFP-BamHI-F: 5’- GGAGGTGGATCCCGTAAAGGAGAAGAACTTTTCA -3’
2. T7T-SpeI-PstI-R: 5’- AAGAGACTGCAGCGGCCGCTACTAGTATATAGTTCCTCCTTTCAG -3’
Cloning Procedure:
The DNA fragment of GFP-His was amplified by PCR and digested by BamHI and PstI, followed by cloning onto Pcons-RBS-LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing. BamHI and PstI, followed by cloning onto Pcons-RBS-LO-BamHI/pSB1C3 which was cut by BamHI and PstI. The part has been confirmed by sequencing.
The AOX gene were cloned onto pSB1C3 driven by a constitutive promoter (BBa_J23101) and RBS (BBa_B0034) and fused with bacterial outer membrane proteins, Lpp-OmpA (LO) to display protein on the cell surface of E. coli. To test whether the gene expression system works, we cloned a reporter gene (GFP) in the same context (i.e., Pcons-RBS-LO-GFP/pSB1C3). And the gene expression level was compared to Pcons-RBS-GFP/pSB1C3 (BBa_K1694035) got from NCTU-Formosa, which has the same promoter and RBS but without LO. The clones of E. coli DH5α were cultured in LB media supplemented with 34 μg/ml chloramphenicol at 37°C overnight. Because overnight cultured LB medium has a background level of fluorescence, the bacterial GFP was measured in the PBS buffer (Enzyme Microb Technol. 2001). As Figure 1 showed, the GFP expression was extremely high in E. coli. LO-GFP fusion protein expression was observed at low but significant level compared to wild-type E. coli or E. coli expressing LO outer membrane proteins.
Figure 1: Gene expression analysis. GFP gene expression was read at Ex/Em = 488/528 nm in BioTek Microplate Spectrophotometer. Lane 1: wild-type E. coli as a mock control; Lane 2: GFP expression in E. coli [BBa_K1694035]; Lane 3: LO outer membrane protein expression [BBa_K1991007] in E. coli; Lane 4: LO-GFP fusion protein expression [BBa_K1991010] in E. coli.
Alcohol oxidase (AOX) was applied in biosensors which catalyzes the oxidation of alcohol followed by hydrogen peroxide production. There are two AOX genes present in a methylotrophic yeast, Pichia pastoris. The gene codon was optimized for expression in E. coli. The DNA sequences were synthesized by IDT and cloned onto pSB1C3 [BBa_K1991000, BBa_K1991001]. In addition, Lpp-OmpA outer membrane proteins, which can direct a fusion protein to display on the bacterial cell surface [BBa_K1991004, BBa_K1991007], were fused with AOX [BBa_K1991005, BBa_K1991006]. In our experimental results, the data showed the enzyme displayed on E. coli can react dose-dependently with various concentration of alcohol in the H2O2 production assay and modeling at the electrochemical analyzer [BBa_K1991002, BBa_K1991003, BBa_K1991008, BBa_K1991009, BBa_K1991010]
Part Name Description Type Length Designer
BBa_K1991000 BBa_K1991000 AOX1/pSB1C3 Basic 1992 bp Chen, Pei-En
BBa_K1991001 BBa_K1991001 AOX2/pSB1C3 Basic 1992 bp Chen, Pei-En
BBa_K1991002 BBa_K1991002 Pcons-RBS-AOX1/pSB1C3 Composite 2053 bp Chen, Pei-En
BBa_K1991003 BBa_K1991003 Pcons-RBS-AOX2/pSB1C3 Composite 2053 bp Chen, Pei-En
BBa_K1991004 BBa_K1991004 Lpp-OmpA-BamHI/pSB1C3 Basic 441 bp Chen, Pei-En
BBa_K1991005 BBa_K1991005 LO-AOX1-His/pSB1C3 Basic 2601 bp Chen, Pei-En
BBa_K1991006 BBa_K1991006 LO-AOX2-His/pSB1C3 Basic 2601 bp Chen, Pei-En
BBa_K1991007 BBa_K1991007 Pcons-RBS-LO-BamHI/pSB1C3 Composite 502 bp Chen, Pei-En
BBa_K1991008 BBa_K1991008 Pcons-RBS-LO-AOX1-His/pSB1C3 Composite 2662 bp Chen, Pei-En
BBa_K1991009 BBa_K1991009 Pcons-RBS-LO-AOX2-His/pSB1C3 Composite 2662 bp Chen, Pei-En
BBa_K1991010 BBa_K1991010 Pcons-RBS-LO-GFP-His/pSB1C3 Composite 1384 bp Chen, Pei-En | {
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-7,296,091,614,355,100,000 | Advertisement
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Tissue and Stage-Specific Distribution of Wolbachia in Brugia malayi
• Kerstin Fischer,
Affiliation: Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America
• Wandy L. Beatty,
Affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America
• Daojun Jiang,
Affiliation: Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America
• Gary J. Weil,
Affiliation: Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America
• Peter U. Fischer
Pufische@DOM.wustl.edu
Affiliation: Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America
Tissue and Stage-Specific Distribution of Wolbachia in Brugia malayi
• Kerstin Fischer,
• Wandy L. Beatty,
• Daojun Jiang,
• Gary J. Weil,
• Peter U. Fischer
PLOS
x
Abstract
Background
Most filarial parasite species contain Wolbachia, obligatory bacterial endosymbionts that are crucial for filarial development and reproduction. They are targets for alternative chemotherapy, but their role in the biology of filarial nematodes is not well understood. Light microscopy provides important information on morphology, localization and potential function of these bacteria. Surprisingly, immunohistology and in situ hybridization techniques have not been widely used to monitor Wolbachia distribution during the filarial life cycle.
Methods/Principal Findings
A monoclonal antibody directed against Wolbachia surface protein and in situ hybridization targeting Wolbachia 16S rRNA were used to monitor Wolbachia during the life cycle of B. malayi. In microfilariae and vector stage larvae only a few cells contain Wolbachia. In contrast, large numbers of Wolbachia were detected in the lateral chords of L4 larvae, but no endobacteria were detected in the genital primordium. In young adult worms (5 weeks p.i.), a massive expansion of Wolbachia was observed in the lateral chords adjacent to ovaries or testis, but no endobacteria were detected in the growth zone of the ovaries, uterus, the growth zone of the testis or the vas deferens. Confocal laser scanning and transmission electron microscopy showed that numerous Wolbachia are aligned towards the developing ovaries and single endobacteria were detected in the germline. In inseminated females (8 weeks p.i.) Wolbachia were observed in the ovaries, embryos and in decreasing numbers in the lateral chords. In young males Wolbachia were found in distinct zones of the testis and in large numbers in the lateral chords in the vicinity of testicular tissue but never in mature spermatids or spermatozoa.
Conclusions
Immunohistology and in situ hybridization show distinct tissue and stage specific distribution patterns for Wolbachia in B. malayi. Extensive multiplication of Wolbachia occurs in the lateral chords of L4 and young adults adjacent to germline cells.
Author Summary
Most filarial nematodes contain Wolbachia endobacteria that are essential for development and reproduction. An antibody against a Wolbachia surface protein was used to monitor the distribution of endobacteria during the B. malayi life cycle. In situ hybridization with probes binding to Wolbachia 16S rRNA were used to confirm results. Only a few cells contain Wolbachia in microfilariae and vector stage larvae; this suggests that the bacteria need to be maintained, but may have limited importance for these stages. Large numbers of Wolbachia were detected in the lateral chords of L4 larvae and of young adult worms, but not in the developing reproductive tissue. Confocal laser scanning and transmission electron microscopy showed that Wolbachia are aligned towards the developing germline. It can be hypothesized that Wolbachia invade developing ovaries from the lateral chords. In inseminated females, Wolbachia were detected in the ovaries and embryos. In young males, Wolbachia were found in parts of the testis and in the lateral chords in the vicinity of testicular tissue but never in mature spermatids or spermatozoa. The process of overcoming tissue boundaries to ensure transovarial transmission of Wolbachia could be an Achilles heel in the life cycle of B. malayi.
Introduction
Filarial parasites infect more than 150 million people in tropical and subtropical countries and are responsible for important tropical diseases such as lymphatic filariasis (elephantiasis) and onchocerciasis (river blindness). Other filarial species are important veterinary pathogens (e.g. Dirofilaria immitis, the dog heartworm). Treatment of filarial infections in humans and animals is suboptimal, because available drugs do not efficiently kill adult worms. Most filarial species live in obligatory symbiosis with intracellular Wolbachia α-proteobacteria. Wolbachia are also present in many insect species, and they are among the most widely distributed bacteria that infect invertebrates. Wolbachia endosymbionts are necessary for development and reproduction of filarial nematodes, and they have been validated as a target for chemotherapy [1]. Tetracycline class antibiotics are active against Wolbachia, and depletion of endobacteria blocks reproduction and eventually kills adult worms in some filarial species [2], [3].
While Wolbachia DNA can be detected and quantified by PCR, microscopy provides important information on morphology and localization of bacteria in parasite tissues. Immunohistochemistry has been used for years to visualize Wolbachia in filarial worms, particularly in Onchocerca volvulus [2]. Brugia malayi is the only human filarial parasite that can be maintained in laboratory animals and for which all life cycle stages are relatively easily accessible. The population dynamics of Wolbachia during the development of B. malayi has been studied by quantitative PCR; for example, the number of Wolbachia exponentially increases soon after infection of the vertebrate host [4]. Recent studies have shown that Wolbachia are unevenly distributed in intrauterine embryos and that the bacteria are not always detected in germline precursor cells [5]. However, data on the histological distribution of Wolbachia during later development of B. malayi are scarce. While it is known that Wolbachia are present in developing embryos, the mechanism of this vertical transmission is poorly understood.
In situ hybridization has been used to study gene expression in filarial parasites such as B. malayi [6] and to detect Wolbachia in insects [7], [8], but it has not been used before to detect Wolbachia in filarial worms. In this paper, we have used optimized immunohistology, in situ hybridization, and transmission electron microscopy to systematically describe the distribution, the relative number and morphology of Wolbachia in different life stages and tissues of B. malayi. This work led to an interesting new hypothesis on the localization and migration of Wolbachia during development of filarial worms.
Materials and Methods
Parasite material
B. malayi worms were recovered from intraperitonial ( i.p.) infected jirds, 2, 5, 8 and 12 wks post infection (p.i.) as previously described [9]. Aedes aegypti mosquitoes containing different larval stages of B. malayi were available from a previous study. Parasite material was fixed either in 80% ethanol for immunohistology or in 4% buffered formalin for immunohistology or in situ hybridization. At least five blocks with four or more B. malayi worms each were examined for each time point. An extensive overview about the studied material and the methods performed is provided in a supplementary table (Table S1). Up to twenty serial sections of the same block were used for comparative studies of different staining procedures. For some blocks (especially those containing young adult worms) more than 60 sections (5 µm) were cut, but only a selection of sections was examined. For the ultrastructural analysis, 18 worms (39 and 56 days p.i., Table S1) were fixed in 2% paraformaldehyde/2.5% glutaraldehyde (Polysciences Inc., Warrington, PA, USA) in 100 mM phosphate buffer, pH 7.2 for 1 hr at room temperature.
Antibodies
A monoclonal antibody directed against the B. malayi Wolbachia surface protein (mab Bm WSP) was purified from culture supernatants kindly provided by Dr. Patrick J. Lammie, Atlanta [10]. Briefly, hybridoma supernatant was incubated overnight at 4°C with ammonium sulfate, pelleted, resuspended in water and dialyzed extensively against phosphate buffered saline. The antibody solution was concentrated to 5% of the original volume using Centricon Plus-20 columns (Millipore, Billerica, MA, USA) and the protein content was determined. A stock mab solution of 10 mg protein per ml was used to test dilution series of 1∶10 up to 1∶500. The best signal to background relationship was observed at a dilution of 1∶100, and this dilution was used for all further experiments.
Immunohistology
The alkaline phosphatase anti-alkaline phosphatase (APAAP) technique was applied for immunostaining according to the recommendations of the manufacturer (Dako, Carpinteria, CA, USA) and as described earlier [11]. TBS with 1% albumin was used as negative control. Rabbit-anti mouse IgG (1∶25; Dako) was applied as secondary antibody and was bound to the APAAP complex. As substrate for alkaline phosphatase the chromogen Fast red TR salt (Sigma) was used and hematoxylin (Merck, Darmstadt, Germany) served as the counter-stain. Sections were examined using an Olympus-BX40 microscope (Olympus, Tokyo, Japan) and photographed with an Olympus DP70 microscope digital camera. For some fluorescent analysis wheat germ agglutinin (WGA 633, Invitrogen, Carlsbad, CA, USA) was used as membrane stain at 200 µg/ml for 10 minutes prior to mounting.
FITC conjugated anti-mouse IgG (1∶300; Sigma) was used as a secondary antibody for confocal laser scanning microscopy (LSM). Sections were examined with a Zeiss LSM 510 META (Zeiss, Jena, Germany) confocal laser scanning microcope equipped with a plan-apochromat 63× oil objective with an argon or helium/neon laser for excitation at 488 nm or 633 nm, respectively. Confocal Z slices of 0.8 µm were obtained using Zeiss LSM software. The Velocity program version 5.4.2 (Improvision, Lexington, MA, USA) was used for high resolution interactive 3D rendering. Sections were also examined using a wide field fluorescence microscope (WFFM, Zeiss Axioskop 2 MOT Plus) with plan-apochromat 100× oil, 63× or 40× objectives. Wide field fluorescence microscopy and LSM were performed at the Washington University Molecular Microbiology Imaging Facility (http://micro.imaging.wustl.edu/).
rRNA probe in situ hybridization
A 424 bp fragment of the 16S rRNA gene of Wolbachia of B. malayi was amplified (forward primer 5′CAGCTCGTGTCGTGAGATGT, reverse primer 5′ CCCAGTCATGATCCCACTT) and cloned into a dual promoter PCRII plasmid (Invitrogen). After linearization of the plasmid, probes (anti-sense) and negative controls (sense) were prepared with Megascript T7 and Sp6 high yield transcription kits according to the manufacturer's suggested protocol (Ambion, Invitrogen). For labeling of the probe a biotin-16 dUTP mix (Roche, Indianapolis, IN, USA) was used during in vitro transcription. The plasmid template was then removed by DNase digestion (Roche). The probes were concentrated by ethanol precipitation, re-suspended in DEPC-treated water, and stored at −20°C until use.
For staining, 5 µm thin paraffin sections were deparaffinized and partially digested with pepsin HCl for approximately 7 minutes. Sections were hybridized at 60°C overnight in a humid chamber with 1 µg of rRNA probe in hybridization buffer (50% formamide, 5XSSC, 0.3 mg/ml yeast tRNA, 100 µg/ml heparin, 1× Denhart's Solution, 0.1% CHAPS and 5 mM EDTA). A stringency wash was performed at 60°C for 30 min, and detection was performed using the ‘In situ Hybridization Detection System’ (K0601, Dako) which uses alkaline phosphatase conjugated streptavidin to localize biotinylated rRNA probes. Sections were incubated for 20 min with streptavidin-AP conjugate at room temperature. BCIP/NBT substrate solution was added for 10 to 30 min to localize binding of the probes.
DNA oligonucleotide probe fluorescence-based in situ hybridization (FISH)
Sections were deparaffinized and partially digested as described above and hybridized at 37°C overnight in a dark humid chamber using 200 ng of a custom made, labeled 30-mer antisense probe targeting the 16S rRNA of Wolbachia (wBm16S as, 5′Alexa 488-CAGTTTATCACTAGCAGT TTCCTTAAAGTC, Invitrogen). The complementary sense sequence was used as a negative control probe. One stringency wash was performed at 37°C for 30 minutes. Hybridization and stringency buffers were the same as described above. Finally sections were rinsed briefly in PBS and covered with a cover slip with ProLong Gold antifade reagent that contains DAPI (Invitrogen). This embedding reagent enables simultaneous fluorescence-based detection of condensed DNA in eukaryotic and prokaryotic organisms. Sections were examined using an Olympus-BX40 microscope equipped with the Olympus fluorescence filter 41001 (excitation 460–500 nm, emission 510–550 nm) for Alexa fluor or UN31000V2 (excitation 325–375 nm, emission 435–485 nm) for DAPI.
Transmission electron microscopy
For ultrastructural analysis fixed samples were washed in phosphate buffer, embedded in agarose, and postfixed in 1% osmium tetroxide (Polysciences Inc.) for 1 hr as described previously [12]. Samples were then rinsed extensively in dH20 prior to en bloc staining with 1% aqueous uranyl acetate (Ted Pella Inc., Redding, CA, USA) for 1 hr. Following several rinses in dH20, samples were dehydrated in a graded series of ethanol and embedded in Eponate 12 resin (Ted Pella Inc.). Sections of 95 nm were cut with a Leica Ultracut UCT ultramicrotome (Leica Microsystems Inc., Bannockburn, IL,USA), stained with uranyl acetate and lead citrate, and viewed on a JEOL 1200 EX transmission electron microscope (JEOL USA Inc., Peabody, MA, USA).
Results
Localization of Wolbachia in larval B. malayi
Different developmental stages were stained with mab Bm WSP (Fig. 1A, C–K). Results were confirmed by in situ hybridization or DAPI chromatin staining. Clusters of Wolbachia were detected in relatively few cells in microfilariae (Fig. 1A). The same staining pattern was observed by rRNA in situ hybridization with a probe for Wolbachia 16S rRNA (Fig. 1B). Wolbachia were sometimes detected in single cells of microfilariae within the midgut of mosquito vectors (Fig. 1C) or in sausage stage larvae and 2nd stage larvae in the mosquito thorax (Fig. 1D, E), but most of the cells in these larval stages were free of the endobacteria. Even in infective 3rd stage larvae the vast majority of cells were devoid of Wolbachia (Fig. 1F); Wolbachia in L3 were mainly present in the cells of the lateral chord, but not in internal organs (Fig. 1G).
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Figure 1. Detection of Wolbachia in larval B. malayi by immunohistology or by in situ hybridization.
A Clusters of Wolbachia (arrow) in single cells of a stretched intrauterine microfilaria labeled by immunohistology using mab Bm WSP. B Consecutive section to A, but Wolbachia (arrow) were detected by in situ hybridization. C Wolbachia (arrow) in single cells of a microfilaria within the midgut of A. aegypti 2 h after the blood meal. D Clustered Wolbachia in a cell of a fragment of an L2 larva in the thorax of A. aegypti 3 d.p.i. E Cross-section of L2 larvae in the thorax of A. aegypti showing a cluster of Wolbachia (arrow) in the hypodermis (7 d.p.i.) F Cross section of isolated infective L3 larvae showing only a single Wolbachia cluster (arrow) in the cells of lateral chord (14 d.p.i). G Several clusters of Wolbachia (arrows) in the lateral chord in a longitudinal section of an infective L3 larva migrating through the abdomen of A. aegypti 14 d.p.i. H Cross-section of the anterior end of a 4th stage larva (14 d.p.i. of a jird). No Wolbachia were detected. I Cross-section of the midbody region of a 4th stage larva showing many Wolbachia (arrow) in one lateral chord. J Cross-section of the midbody region of another 4th stage larva (wide field fluorescence microscopy) showing highly condensed DNA (blue DAPI stain) in the genital primordium and numerous Wolbachia (arrow) in the lateral chord similar to I. K Longitudinal section of the midbody region of a 4th stage larva with many Wolbachia in the lateral chords (arrows). Cells in the pseudocoelomic cavity were free of Wolbachia. Ph, pharynx; gp, genital primordium; I, intestine; m, muscle. Scale bar 10 µm.
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The Wolbachia density at the anterior end of 4th stage larvae was low at 2 weeks p.i. in the vertebrate host (3–5 days after the molt), and no endobacteria were detected in the tissue around the pharynx (Fig. 1H). In contrast, large numbers of Wolbachia were detected in the developing lateral chords of the L4 midbody region (Fig. 1I–K).
Detection of Wolbachia during the development of adult female B. malayi
In order to understand the distribution of Wolbachia in adult worms it is crucial to recall the anatomy and development of reproductive organs of filarial worms [13], [14]. The genital opening (vulva) lies close to the anterior end of the female worm, approximately at the level of the esophagus (Fig. 2A, B). The vagina leads into the bifurcated uterus which ends in the seminal receptacles. Theses organs are linked by oviducts with two ovaries that have an anterior growth zone, a maturation zone in the middle, and a posterior germinative zone. At 5 weeks p.i. in the vertebrate host, young adult female B. malayi worms are approximately 1.8 cm long and still growing. At that time point a massive accumulation of Wolbachia was observed, mainly in the lateral chords. Increased numbers of Wolbachia were observed in the lateral chords in the posterior end of the female which was still free of ovaries (Fig. 3A). Sections of the posterior part of the ovaries showed large numbers of Wolbachia in the adjacent lateral chords, but the ovaries themselves were free of Wolbachia (Fig. 3B, C, 4A). In the oocyte maturation and growth zones of the ovaries, Wolbachia were oriented within the lateral chords towards the pseudocoelomic cavity (Fig. 3D–G), and some sections showed Wolbachia in the periphery of the ovary (Fig. 3H, 4B, D, F). Two distribution patterns of Wolbachia were found in the lateral chords. Scattered Wolbachia were present in the apical part of the chords, and numerous clusters of Wolbachia were present basal border of the hypodermal chords adjacent to the ovaries (Fig. 3I). A similar staining pattern for Wolbachia was observed in the lateral chords in the midbody region of 5 week old females, but their empty uterus branches were always free of Wolbachia (Fig. 3J). In 8 week old female worms, less Wolbachia were detected in the lateral chords, but the mature ovaries in the posterior part of the worms were heavily infected with Wolbachia (Fig. 5A). These worms contained developing microfilariae, and the ovaries showed strong staining of nuclear chromatin (as determined by DAPI). Morula stage embryos were observed in the uterus with many Wolbachia, while in this region the number of endobacteria in the lateral chords was lower than in the distal parts of the lateral chords.
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Figure 2. Schematic drawing the anatomy of adult stage B. malayi and distribution of Wolbachia.
Hypodermis, muscles, median chords as well as nerve and secretory-excretory system are not shown. Proportions are estimates. Tissues and organ systems are simplified and the midbody region is interrupted for clarity. The lateral chords are shown dorsally and ventrally instead of laterally. A Adult microfilaria producing female (12 weeks p.i.). The body length is about 4 cm. Wolbachia (red dots) are localized mainly in the lateral chords, ovary and developing embryos. The lateral chords in the head (up to the vulva) of the worm rarely contain Wolbachia. B Adult stage, immature female (5 weeks p.i.) a few days after the 4th molt. The body length is approximately 1.8 cm. Wolbachia are mainly localized in the lateral chords. Occasionally Wolbachia are attached to the ovaries or single endobacteria can be found within the ovary. C Adult spermatozoa producing male worm (12 weeks p.i.) with a total body length of about 2.5 cm. Spiculae are not shown. Wolbachia are localized in the lateral chords; Wolbachia remnants can be detected in parts of the vas deferens but not within spermatids or spermatozoa.
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Figure 3. Detection of Wolbachia in immature female B. malayi at 5 weeks p.i.
AD, HJ immunohistology using mab Bm WSP. A Cross-section in the posterior part of the female showing massive accumulation of Wolbachia (arrow) in the lateral chord. B Longitudinal section of the distal tip showing Wolbachia-free growing ovaries and numerous clusters of Wolbachia (arrow) in the lateral chord. C Cross-section showing asymmetric distribution of large amounts of Wolbachia (arrow) in the lateral chord and Wolbachia-free ovaries. D Large numbers of Wolbachia (arrow) in close proximity to one ovary branch. Ovaries are still not infected. EG Consecutive sections to D. E 16S rRNA in situ hybridization. F 16S oligonucleotide FISH. G DAPI stain showing highly condensed chromatin in the ovary and in Wolbachia in the lateral chord (arrow). H Cross-section showing not only Wolbachia clusters in the lateral chords (arrow) but also at the edge of one ovary (arrow head). I Sagital section through lateral chord and ovary showing large numbers of Wolbachia clusters (arrows) in the lateral chords aligned at the ovary and some Wolbachia clusters in the middle of the chord (arrow head). J Cross-section of the midbody region showing large numbers of Wolbachia in the lateral chord (arrow) and empty uterus branches. i, intestine; u, uterus; ov, ovary. Scale bar 25 µm.
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Figure 4. Detection of Wolbachia in female B. malayi at 5 weeks p.i. by advanced microscopy techniques.
Confocal laser scanning microcopy (LSM) and wide field fluorescence microscopy (WFFM) were used together with mab Bm WSP (green) to label Wolbachia and a WGA 633 stain (red) to visualize membranes. WFFM contained also a filter for DAPI to detected concentrated DNA. A Cross-section showing large amounts of Wolbachia in the lateral chords close to the ovaries (arrow). Some Wolbachia were also detected in the hypodermis. For full scans of LSM images and 360° rotation see suppl. material (LSM). B Another cross-section showing a more advanced stage of infection with endobacteria attached to the ovary membrane (arrow head) and already in the ovaries (arrow) (LSM). C Longitudinal section showing Wolbachia lining up in the lateral chords in the vicinity of the ovary (arrow) or within one ovary branch (arrow head)(LSM). D WFFM showing Wolbachia approaching ovaries with highly condensed chromatin (blue DAPI stain). E Another cross-section showing Wolbachia (arrows) attached to the ovaries (blue DAPI stain). One ovary branch is already Wolbachia (arrow head) infected while the other is still free (WFFM). F Close-up of a longitunial section showing Wolbachia (arrow) directly at the ovary membrane (red) (WFFM). G Another close-up showing Wolbachia in the lateral chords close to the ovary (arrow) or already in the somatic cells of the ovary (arrow head) (WFFM). Staining for WSP provides a characteristic donut shaped pattern of a Wolbachia surface protein. Ov, ovary; i, intestine. Scale bar 30 µm.
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Figure 5. Detection of Wolbachia in adult female B. malayi at 8 and 12 weeks p.i.
In A, B, E, F and H Wolbachia were labeled by mab Bm WSP. A Cross-section of a distal end of a worm 8 weeks p.i. showing Wolbachia infected ovaries and some Wolbachia clusters in the lateral chord. BH B. malayi 12 weeks p.i. B Several cross-sections of the ovaries showing massive Wolbachia staining (arrow), but only weak labeling in the adjacent lateral chord (arrow head). C Consecutive section to B, but in situ 16S rRNA detection confirmed the large number of Wolbachia in the ovaries with much stronger labeling in the adjacent lateral chord (arrow head). D Cross-section of the midbody region shows both uterus branches full of morula stage embryos and Wolbachia (arrow, 16S rRNA). E Longitudinal section in the region of the transition of the ovaries to the tubular oviduct. Large numbers of Wolbachia are visible in the growth zone of the ovaries (arrow heads) and the adjacent lateral chords (arrow) but not in the oviduct. F Cross-section showing one uterus branch with Wolbachia positive (arrow) morula stage embryos and numerous spermatozoa in the seminal receptacle surrounding a few egg cells. Thin lateral chords show only scattered Wolbachia. G Consecutive section to F stained with polyclonal Brugia pahangi WSP antisera [39] shows Wolbachia in the egg cells (arrowhead). H Cross-section of the region close to the female genital opening. Some Wolbachia are visible in the lateral chord (arrow) and in the stretched microfilariae (arrowhead) in the vagina uterine, but the muscular vagina vera is free of microfilariae and Wolbachia. Ov, ovary; u, uterus; od, oviduct; sp, spermatozoa; vv, vagina vera; vu, vagina uterina. Scale bar 25 µm.
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In 12 week old females numerous Wolbachia were observed in the lateral chords and the posterior parts of the ovaries, but bacteria densities in these areas were lower than in the lateral chords adjacent to the anterior ovary and oviduct (Fig. 5B,C,E, H). Numerous Wolbachia were detected in morula stage embryos in the uterus, but only a few were detected in the lateral chords of females at that level (Fig. 5D, F, G). Intrauterine spermatozoa surrounding degenerated oocytes in the seminal receptacle were free of Wolbachia, but serial sections showed some Wolbachia in the oocytes in this area (Fig. 5F,G). Stretched microfilariae in the vagina uterina contained Wolbachia in some cells, but the numbers were low compared to those in morula stage embryos. This suggests that Wolbachia may be necessary for rapid cell division which occurs in developing embryos but not in stretched microfilariae. The distribution of Wolbachia in the lateral chords was often asymmetrical, and this depended on the proximity to the reproductive system, body region and on the age of the worm.
Detection of Wolbachia during the development of adult male B. malayi
The genital opening of the male worm lies at the posterior end and forms with the anus a cloaca (Fig. 2C). This is in stark contrast to the anatomy of females. A single vas deferens leads into a seminal vesicle that is connected to the testis; this can be subdivided into a growth zone, a maturation zone, and a germinative zone. In parallel to the distribution of Wolbachia in females, large numbers of endobacteria were observed in the lateral chords of 5 week old males, while the growing sections of the testes in the midbody region were free of Wolbachia (Fig. 6A). However, Wolbachia were present in 5 week males near the testes (Fig. 6B–E) and in the middle part of the testis itself (Fig. 6 F–J). No Wolbachia were detected within the vas deferens by immunohistology (Fig. 7A, D). In contrast, Wolbachia 16S rRNA was detected by in situ hybridization in the testis tissue surrounding the spermatocytes and in the periphery of the vas deferens that contained spermatids (Fig. 7B, C, E). Wolbachia were never observed in the spermatids or the spermatozoa.
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Figure 6. Detection of Wolbachia in immature male B. malayi at 5 weeks p.i.
In A and C Wolbachia were labeled by mab Bm WSP). A This cross-section of the midbody region shows primary spermatogonia in the testis and numerous Wolbachia (arrow) in the lateral chords. B LSM of a consecutive section of A. Individual clusters of Wolbachia (arrow) can be identified. C Two cross-sections more distal compared to A with Wolbachia in the lateral chords (arrow) and in tissue attached to the testis (arrow head). D Consecutive section to C examined by LSM. E Consecutive section to C stained by in situ rRNA hybridization confirming the staining pattern. FJ Serial cross-sections showing many Wolbachia in the lateral chords (arrowheads) and in the spermatogonia (arrows). F mab Bm WSP. G Also mab Bm WSP but LSM H Same staining as G but WFFM with DAPI stain of testis. I 16S in situ rRNA hybridization. J 16S oligonucleotide FISH. te, testis. Scale bar 25 µm.
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Figure 7. Detection of Wolbachia in male B. malayi at 12 weeks p.i.
In A and D Wolbachia were labeled by mab Bm WSP). A Two cross-sections demonstrating Wolbachia in the lateral chords (arrows). One section is in the midbody region showing developed spermatogonia and numerous Wolbachia in the lateral chord, while the other section is in the more muscular distal end of the worm showing spherical spermatids in the vas deferens and fewer Wolbachia in the lateral chord. B Consecutive section to A, but stained with 16S rRNA in situ hybridization. In contrast to A some staining for Wolbachia rRNA is also detected in the testis (arrowheads) and at the border of the vas deferens (arrow heads). C Cross-section of the distal part of another male worm showing Wolbachia 16S rRNA labeling at the epithelium of the vas deferens (arrow heads) and in the lateral chord (arrow). Spermatozoa were never labeled. DG Consecutive cross-section through the terminal end showing the vas deferens containing fully developed spermatids in transition to spermatozoa. Wolbachia are detected in the lateral chord (arrow), but not in the spermatids. D mab Bm WSP. E 16S rRNA in situ hybridization. Similarly to B, staining is also observed at the border of the vas deferens (arrowheads). F 16S oligonucleotide FISH. Although Wolbachia can be easily identified in the lateral chord, granular staining at the membrane of the vas deferens is difficult to recognize. G DAPI stain, again Wolbachia can be easily identified in the lateral chord, but granular staining at the membrane of the vas deferens is hard to differentiate. Sp, spermatozoa; te, testis; vd, vas deferens. Scale bar 25 µm.
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Comparison of morphological detection methods
Comparison of four different methods on consecutive sections (Figs. 3 D–G; 6 C–E; 6 F–J; 7A,B; 7D–G) revealed almost identical staining patterns for Wolbachia by immunohistology with mab Bm WSP, by in situ hybridization (using FISH and RNA in situ to detect Bm Wolbachia 16S rRNA), and by DAPI staining. Differences between immunohistology and 16S rRNA in situ detection were occasionally observed (Figs. 5B, C; 7A, B; 7D, E). In these cases in situ hybridization detected a strong Wolbachia 16S rRNA signal, while no or very little Wolbachia surface protein was detectable by immunohistology. This may indicate a small difference of gene expression pattern or of gene product stability of both markers, but was not noticed as confounding factor. In addition, the intestine of B. malayi was sometimes nonspecifically labeled by immunohistology because of endogenous alkaline phosphatase (e.g. Fig. 5A, F–H). This did not occur with in situ staining.
The mab Bm WSP immunohistology assay detects a protein on the surface of Wolbachia, and it is possible that this protein is not present on all Wolbachia cells. In contrast, the in situ hybridization assay detects expression of 16S rRNA in the cytoplasm of Wolbachia. Small subunit rRNA is known to be highly expressed during the exponential growth phase of bacteria, and that has been used as marker for viability [15]. Therefore the in situ assay is an excellent marker for Wolbachia growth, and it may be suitable for assessing both the presence and viability of Wolbachia. DAPI staining, which detects A-T rich regions in DNA, is an easy and quick method to detect Wolbachia in the lateral chords, since this syncytial tissue usually does not contain condensed filarial chromosomes (Figs. 3G; 7G). However, it is difficult to identify Wolbachia by DAPI staining in areas with condensed filarial chromosomes such as ovaries or in spermatids within the vas deferens (Figs. 3G; 7G). This problem can be solved by combining the DAPI stain for condensed DNA with immunohistology (Figs. 2J, 4D–G; 6H). This permits visualization of Wolbachia in the vicinity of filarial nuclei.
Confocal laser scanning microscopy
Confocal laser scanning microscopy was used to study the three dimensional distribution of Wolbachia in larvae and in developing reproductive tissue of young adult worms. Although Wolbachia numbers were increasing in the lateral chords in 4th stage larvae, no Wolbachia were observed in developing reproductive organs in L4. The higher resolution of LSM confirmed heavy Wolbachia loads in the lateral chords of young female worms (5 weeks) and relatively few endobacteria in the hypodermis (Fig. 4A). Entire oocytes could be examined for Wolbachia, because the size of oocytes is less than 5 µm and the scanned slices were 0.8 µm thick which is about the size of an endobacteria. The confocal examination of the distal end of the ovaries in 5 week old females confirmed the absence of Wolbachia from primary oocytes (Fig. 4A, D). A full LSM scan and rotation of the section show that Wolbachia were present also in the hypodermal pouches that form longitudinal lines in 5 week old female worms (video S1). A membrane stain helped to demonstrate that some Wolbachia were attached to the external membrane around the proximal ovary while other bacteria were actually in the ovary (Fig. 4B, C, videos S2, S3). The latter Wolbachia were always in the vicinity of large clusters of Wolbachia in the lateral chords adjacent to the ovaries in developing adult female worms (Fig. 4B, C). Wide field fluorescence microscopy using FITC labeled mab Bm WSP with a membrane stain and an overlay of the DAPI nuclear stain showed that Wolbachia are attached to the ovary membranes (Fig. 4D, E, F, G). It is possible that these endobacteria invade the ovaries of young females from the lateral chords. Wolbachia distribution in the developing ovaries was not uniform; in some cases, one branch was infected while the other branch was Wolbachia free (Fig. 4E).
Ultrastructural studies of Wolbachia in developing reproductive tissue
Studies of the midbody region of 5 week old worms by transmission electron microscopy confirmed the presence of Wolbachia in the vicinity of developing reproductive tissues. Numerous rod-shaped and spherical Wolbachia were detected in the lateral chords in females, especially in adult worm tissues that are adjacent to developing ovaries. In some areas the hypodermal chord tissue was loose and vacuolized (Fig. 8A). The epithelial cells surrounding the basal lamina of the ovaries were occasionally also strongly vacuolized indicating tissue degeneration, and small, electron dense Wolbachia were detected in these vacuoles (Fig. 8B). Occasionally extracellular Wolbachia were seen in the pseudocoelomic cavity docking to the edge of the ovaries (Fig. 8C, D) or attached to the outer ovarian tissue (Fig. 8E, F). While most of the Wolbachia in the lateral chords were rod-shaped or spherical and up to 1 µm in length and 0.5 µm in diameter, the endobacteria in the pseudocoelomic cavity were condensed, bacillary in shape and only 0.15 to 0.5 µm in length (Fig. 8G–I). Within the ovaries, these small Wolbachia forms were observed in large vacuoles or in loose ovarian tissue (Fig. 8G, I) either as single bacteria or in groups (Fig. 8H).
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Figure 8. Transmission electron microscopy of young adult female B. malayi 5 weeks p.i.
A Numerous Wolbachia (arrow) in the lateral chord in the vicinity of developing ovaries. A number of small, electron dense Wolbachia can be detected in more loosened tissue (arrow heads). B Vacuolized epithelium surrounding the ovary with electron dense Wolbachia (arrow heads) on both sides of the basal lamina. C Numerous Wolbachia in the lateral chord (arrow) and a bacilli-shaped, extracellular Wolbachia in the pseudocoelomic cavity (arrow head). D Magnification of C, showing the Wolbachia cell membrane (arrow head). E Wolbachia (arrow head) docking to the outer ovary epithelium. F Magnification of the endobacterium in E showing loosened cell membrane at the apical end (arrow head) and a dense central inclusion (arrow). G Small electron dense Wolbachia (arrow heads) of the oogonia in the proximal part of the ovary. H Single and clusters of electron dense, extracellular Wolbachia (arrow heads) in the ovary. I Electron dense Wolbachia (arrow heads) in vacuolized ovary tissue. Bl, basal lamina; lc, lateral chord nu, nucleus; ov, ovary; ps, pseudocoelomic cavity. Scale bar 0.5 µm.
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In 5 week old male worms large clusters of large, rod-shaped or spherical Wolbachia were observed in the lateral chords in the vicinity of the testis (Fig. 9A). Small, bacillary Wolbachia forms were sometimes observed in the testis tissue. At the caudal end of the testis, close to the transition to the vas deferens, Wolbachia were observed in the inner tissue, sometimes in the vicinity of peripheral spermatids (Fig. 9B,C, D). These spermatids can be easily identified and differentiated from mature spermatozoa by their compact membranous organelles and the absence of major sperm protein complexes. Large amounts of membranous material were observed in the lumen between the spermatids and the inner testis epithelium. This material resembles degenerating Wolbachia (Fig. 9B, E–G) as they have been described previously [16]. Wolbachia were unambiguously identified in the reproductive tissue of young male worms, but not in the spermatids or spermatozoa.
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Figure 9. Transmission electron microscopy of young adult male B. malayi 5 weeks p.i.
A Numerous Wolbachia (arrows) are observed in the lateral chords close to the testis. Single dark Wolbachia (arrow head) are found in the testis close to the membrane. Mitochondria (asterisk) are found in the periphery of the lateral chord. B Wolbachia (arrow) in the inner testis epithelium in the vicinity of a spermatid. Large amounts of membranous material (arrow heads) can be observed in the testis lumen in the vicinity of the testis epithelium. C Magnification of B showing intracellular Wolbachia (arrow). D Another sample showing Wolbachia (arrows) in the inner testis epithelium. EG Pleomorphic Wolbachia (arrows) in vacuolized testis tissue. Membranous material (arrow head) can be seen in extracellular spaces. Ps, pseudocoelomic cavity; lc, lateral chord; sp, spermatide; mo, membranous organelle; nu, nucleus; bl, basal, lamina; te, testis. Scale bar 0.5 µm.
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Discussion
Immunohistology has been extensively used to study Wolbachia and their clearance following chemotherapy in O. volvulus. Compared to O. volvulus, mature B. malayi have a thinner hypodermis and less pronounced lateral chords, and this can make the detection of Wolbachia more difficult. Our results demonstrate that the distribution and density of Wolbachia vary in different tissues and developmental stages. Our results are consistent with those from a PCR study that reported low amounts of Wolbachia DNA in vector stages and larger amounts in mammalian stages [4]. McGarry and co-workers reported an exponential increase in Wolbachia DNA in transmitted B. malayi L3 larvae as early as 7 days p.i. We detected large amounts of endobacteria by histology in the lateral chords of the midbody region in L4 larvae (14 d.p.i.). More Wolbachia were present in young adult worms at 35 d.p.i. in most parts of the lateral chords and also in an uneven distribution in the hypodermis.
Observations on Wolbachia density and tissue localization may lead to hypotheses regarding their potential function in filarial worms. Antibiotic treatment experiments have suggested that Wolbachia may play a crucial role in the molting process of filarial parasites [17], [18], [19], [20]. It appears clear that if Wolbachia have a direct function during molting, this function does not require localization in the vicinity of the filarial cuticle, since our localization results show that Wolbachia are not located near the cuticle during or immediately after molting. The distinct age and tissue specific distribution patterns of Wolbachia suggest also that the bacteria are not likely to be needed for housekeeping functions in all cell types of filarial nematodes. The absence of Wolbachia in the filarial nervous system, muscles, or the digestive systems suggests that Wolbachia are not needed for these functions. In adult worms the majority of mitochondria can be found in the periphery of the lateral chords, while the majority of Wolbachia are localized in or near the reproductive system. The differential distribution of Wolbachia and mitochondria within the lateral chord of filarial parasites has been reported previously [21]. Especially to the female worms the localization of Wolbachia in the lateral chords in vicinity of the reproductive system implies an important role of endobacteria for embryogenesis and intrauterine development. In agreement with this hypothesis tetracycline treatment to deplete Wolbachia in developing filarial worms has been shown to affect mainly females and causes a male-biased sex-ratio [20], [22].
The Wolbachia genome in B. malayi encodes complete pathways for the biosynthesis of nucleotides, riboflavin, flavin adenine dinucleotide and heme, which are missing or incomplete in the filarial genome [23]. A high demand for gene products (which may not be taken up from the mammalian host) from these pathways might be especially necessary during the development of the reproductive system in young adult worms. Furthermore, the phylogenetically old and tight association of filarial nematodes with Wolbachia during reproduction may have led to additional interdependencies that account for their mutualistic relationship. As hypothesized for Wolbachia in insects, it is possible that Wolbachia in filarial nematodes are especially important for pre-meiotic mitosis, meiosis, and meiosis associated processes [24], [25], [26], [27].
A recent study examined the dynamics of Wolbachia during intrauterine embryogenesis of B. malayi using Caenorhabditis elegans embryogenesis as a framework for the analysis [5]. Asymmetric Wolbachia segregation was observed that could explain the concentration of Wolbachia in the hypodermal chords. The early differential distribution of Wolbachia within embryonic cells corresponds well with the strong tissue specific distribution in later development described in our study. However, the authors also hypothesized that the asymmetric segregation pattern may be responsible for the presence of Wolbachia in the female germline [5]. This is in contrast to our results which clearly demonstrate the absence of Wolbachia in male and female reproductive tissue from the third stage larvae to the young adult worms. Since it is difficult or impossible to identify the germline cells or gender of microfilariae, vector stage first stage larvae, and second stage larvae of B. malayi, we cannot be sure when during development Wolbachia are lost in these cells.
The terminal ends of Brugia ovaries form the germinative zones which contain the mitotic growing oogonia [28]. Our study showed that these areas were free of Wolbachia in growing, young adult worms. Our results suggest that Wolbachia from adjacent lateral chords may cross tissue zones to infect cells in maturation zone 1 (which mainly contains primary oocytes in the pachytene stage of meiotic prophase I) and in maturation zone 2 (which contains oocytes in the remaining phases of meiosis I). The germinative zones of the ovaries seem to be populated by Wolbachia over a period of approximately three weeks following the L4–L5 molt. Large numbers of Wolbachia were present in the maturation zones of eight week or older female worms, while the attached growth zones which contain the secondary oocytes and the oviducts contained lower numbers of Wolbachia (see Fig. 5E). Fertilization precedes meiosis II in filarial nematodes [28]. Wolbachia were detected in secondary oocytes surrounded by spermatozoa and unfertilized oocytes within the seminal receptacle in mature females (see Fig. 5F, G).
The picture was similar in male worms. Wolbachia were not observed in the germinative zone of the testis. It is possible that Wolbachia from the lateral chords infect the primary spermatocytes in maturation zone 1, which are mostly in the pachytene stage of prophase of meiosis I [29]. This report is the first detection of Wolbachia in primary spermatocytes of developing male filarial nematodes. Although mature male worms have been previously examined for Wolbachia, prior studies did not report infection of the testis [30]. This is not contradictory to our findings, since Wolbachia appear to only infect the testis of immature adult stage B. malayi males and such worms were not studied previously. The spermatocytes of the adjacent growth zone and maturation zone 2 are difficult to differentiate morphologically, but larger secondary spermatocytes that have completed meiosis and the spherical spermatids which enter the vas deferens can be distinguished. Wolbachia were never seen in the spermatids or the mature spermatozoa. However, our in situ hybridization results clearly indicated the presence of Wolbachia 16S rRNA in the periphery of the seminal vesicle. This was confirmed by electron microscopy that showed Wolbachia in the inner epithelium of the testis or vas deferens, but not in the spermatids. These data may suggest that high Wolbachia densities are correlated with condensed chromatin and Wolbachia may be involved in chromosome segregation of filarial nematodes.
Our ultrastructural studies of young adult B. malayi confirm that Wolbachia are highly pleomorphic. This pleomorphism was recognized shortly after the discovery of endobacteria in filarial nematodes, and it has been suggested that Wolbachia may have a Chlamydia-like life cycle with small dense bodies as potential infectious forms [30], [31]. Chlamydia and filarial Wolbachia both have an obligatory intracellular life style and a small genome size due to the loss of a number of essential biosynthetic pathways. Both bacterial groups lack cell walls but retained a functional lipid II biosynthesis pathway [32]. It is also possible that Wolbachia share the requirement of Chlamydia for host cell sphingolipids supplied by the host cell Golgi apparatus and multivesicular bodies for activation [33]. Clearly, further studies are needed to assign functions to different morphological forms of Wolbachia during the filarial life cycle.
Based on our results we hypothesize that the genital primordium in larval B. malayi is devoid of Wolbachia and that reproductive tissues in young adult worms become infected with Wolbachia from adjacent lateral chords which have many Wolbachia. Prior studies have shown that newly introduced Wolbachia can cross several tissue planes and infect the germline in Drosophila [34]. This could be also the case in filarial Wolbachia, and it is possible that similar host signals trigger the germline tropism of Wolbachia in filarial worms and Drosophila. Previous studies have shown that a Wolbachia htrA serine protease can be found outside bacterial cells in filarial parasites. This protease and other secreted bacterial proteins may be involved in tissue invasion [35]. In addition to tissue lysis, motility of Wolbachia may be necessary for the bacteria to cross tissue boundaries. Actin-based motility occurs in Rickettsia and many other intracellular bacteria [36]. Orthologs of genes essential for actin-based motility have been found in the Wolbachia genome. Additional work will be needed to study the localization and timing of expression for these genes [23], [37].
Our ultrastructural results confirmed the presence of large clusters of Wolbachia in the lateral chords in the vicinity of the ovaries and in the outer ovary epithelium as previously described [30], [38]. The new finding reported here, is the detection of extracellular Wolbachia in the pseudocoelomic cavity in young females and the presence of Wolbachia in testis of developing male worms. In summary, this study shows the value of histological techniques such as immunohistology and in situ hybridization to study the tissue distribution of Wolbachia during the life cycle of filarial nematodes. Wolbachia infection was found to be highly cell and tissue specific. No Wolbachia were found in the developing reproductive organs in fourth stage larvae and freshly molted adult worms, which had heavy Wolbachia loads in the lateral chords. Wolbachia were detected in reproductive tissues with the onset of oocyte and sperm development, and infection of oocytes results in transovarial transmission of Wolbachia to the next generation.
Supporting Information
Table S1.
Summary table for the parasite material used for the present study. Each slide was thoroughly examined and numerous pictures were taken. If a slide contained more than one block section, all sections were analyzed.
doi:10.1371/journal.pntd.0001174.s001
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Video S1.
Full rotation of a cross-section of a 5 week old female B. malayi. Wolbachia were detected in the hypodermal lateral chords and the hypodermis. Confocal laser scanning microscopy stained with mab Bm WSP and WGA 633 (red) to visualize membranes. Compare Fig. 6A.
doi:10.1371/journal.pntd.0001174.s002
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Video S2.
Similar to video S1 but Wolbachia are already attached to the membrane of the one ovary branch. Note these attached bacteria can be noticed only from one side of the section. A few additional endobacteria are already in the ovary. The second ovary branch is still devoid of Wolbachia. Compare Fig. 6B.
doi:10.1371/journal.pntd.0001174.s003
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Video S3.
Longitudinal section of a 5 week old female B. malayi. Wolbachia are lining up at the lateral chords and some Wolbachia in one ovary branch. Confocal laser scanning microscopy stained with mab Bm WSP and WGA 633 (red) to visualize membranes. Compare Fig. 6C.
doi:10.1371/journal.pntd.0001174.s004
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Acknowledgments
We would like to thank Pat Lammie, CDC, for providing the mab Bm WSP cell culture supernatant and Kurt Curtis, Washington University School of Medicine, for his assistance in antibody purification. Yuefang Huang, Washington University, provided B. malayi worms. Marcy Hartstein, Washington University, and Odile Bain, Muséum National d'Histoire Naturelle, Paris, helped with the schematic drawing. We thank Norbert W. Brattig and especially the late Dietrich W. Büttner (both Bernhard Nocht Institute for Tropical Medicine, Hamburg) for valuable discussions.
Author Contributions
Conceived and designed the experiments: KF WLB GJW PUF. Performed the experiments: KF WLB. Analyzed the data: KF WLB DJ GJW PUF. Contributed reagents/materials/analysis tools: WLB DJ. Wrote the paper: KF GJW PUF.
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-3,749,794,889,148,391,400 | ePrints@IIScePrints@IISc Home | About | Browse | Latest Additions | Advanced Search | Contact | Help
Expression of functional aromatase in the epididymis: Role of androgens and LH in modulation of expression and activity
Shayu, D and Rao, AJ (2006) Expression of functional aromatase in the epididymis: Role of androgens and LH in modulation of expression and activity. In: Molecular and Cellular Endocrinology, 249 (1-2). pp. 40-50.
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Abstract
The primary source of 17beta-estradiol (E2) in the male is the testis, which expresses the enzyme complex aromatase that is involved in E2 biosynthesis. However, recent evidences suggest that the epididymis is also capable of E2 biosynthesis. Our results demonstrate the presence of cytochrome P450 aromatase $(P450_A_R_O_M)$ and 17beta-hydroxysteroid dehydrogenase I messenger ribonucleic acid (mRNA) in the caput and cauda regions of rat epididymis. The androgenic substrates testosterone and androstenedione could be utilized by the rat epididymal aromatase for E2 biosynthesis as assessed by radioimmunoassay. $(P450_A_R_O_M)$ expression is transcriptionally regulated in a tissue-specific manner by various factors including androgens and luteinizing hormone (LH). Androgens could positively modulate epididymal $(P450_A_R_O_M)$ mRNA levels as assessed by castration studies, treatment with flutamide or in vitro incubation of tissue minces with 5alpha-dihydrotestosterone (DHT). Several extra-gonadal tissues including the epididymis are known to express LH receptors (LHR). Our study revealed a higher level of LHR mRNA expression in the cauda region compared to the caput. Caudal membrane extracts could bind human chorionic gonadotropin (hCG), which resulted in the production of cAMP. Interestingly, hCG could also regulate $(P450_A_R_O_M)$ mRNA expression in vitro and enhance E2 biosynthesis. Together our results highlight the presence of a functional aromatase in the epididymis that is subject to regulation by LH and androgens.
Item Type: Journal Article
Related URLs:
Additional Information: Copyright of this article belongs to Elsevier.
Keywords: 17β-HSDI;LH receptor;Flutamide;Estradiol biosynthesis
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 27 May 2006
Last Modified: 19 Sep 2010 04:27
URI: http://eprints.iisc.ernet.in/id/eprint/6950
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RN207759 Lhx1 (untagged ORF) - Rat LIM homeobox 1 (Lhx1), (10 ug), NM_145880.3, 10ug $380 3 weeks
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OriGene Data
Vector:pCMV6 Entry QM Insert Size: 1221 Restriction Site: SgfI-MluI
Sequence Data: Fully Sequenced ORF
OTI Disclaimer: Our molecular clone sequence data has been matched to the reference identifier above as a point of reference. Note that the complete sequence of our molecular clones may differ from the sequence published for this corresponding reference, e.g., by representing an alternative RNA splicing form or single nucleotide polymorphism (SNP).
Product Components: The cDNA clone is shipped in a 2-D bar-coded Matrix tube as dried plasmid DNA. The package also includes 100 pmols of both the corresponding 5' and 3' vector primers in separate vials. Every lot of primer is tested to provide clean sequencing of OriGene TrueClones.
Reference Data
RefSeq: NM_145880.3, NP_665887
RefSeq Size: 1938 RefSeq ORF: 1221
Synonyms :
LocusID: 257634 Cytogenetic: 10q26
Summary: may play a role in the regulation of genes in specific neuronal subtypes [RGD, Feb 2006]. ##Evidence-Data-START## Transcript exon combination :: S71523.1 [ECO:0000332] RNAseq introns :: single sample supports all introns SRS214101, SRS369727 [ECO:0000348] ##Evidence-Data-END##
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概述
性能
应用
Our Abpromise guarantee covers the use of ab70747 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用 Ab评论 说明
WB 1/2000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 50 kDa).
IP Use at 2-5 µg/mg of lysate.
靶标
• 功能
Relays signals from the cell surface to the nucleus to weaken adherens junction and promote actin cytoskeleton reorganization and cell invasiveness. Involved in lysophosphatidic acid-induced cell adhesion and migration. Acts as a transcriptional coactivator for NF-kappa-B and JUN, and mediates the transrepression of these transcription factors induced by glucocorticoid receptor.
• 组织特异性
Abundantly expressed in kidney, liver and lung. Lower levels in heart, placenta and pancreas. Expressed in colonic epithelial cells. Up-regulated in colonic tumors.
• 序列相似性
Belongs to the zyxin/ajuba family.
Contains 3 LIM zinc-binding domains.
• 结构域
The LIM zinc-binding domains mediate interaction with LPAR2 and with S.typhimurium protein sseI.
• 翻译后修饰
Phosphorylation at Tyr-55 by SRC is required for enhancement of lysophosphatidic acid-induced cell migration. Tyr-55 is dephosphorylated by PTPN13.
• 细胞定位
Cytoplasm > cytoskeleton. Cell junction > focal adhesion. Nucleus. Cytoplasm. Shuttles between nucleus and cytoplasm.
• Information by UniProt
• 数据库链接
• 别名
• MGC10556 antibody
• MGC10558 antibody
• MGC29959 antibody
• MGC3837 antibody
• MGC4423 antibody
• OIP-1 antibody
• OIP1 antibody
• OPA interacting protein 1 antibody
• OPA-interacting protein 1 antibody
• Thyroid hormone receptor interactor 6 antibody
• Thyroid receptor interacting protein 6 antibody
• Thyroid receptor-interacting protein 6 antibody
• TR-interacting protein 6 antibody
• TRIP-6 antibody
• Trip6 antibody
• TRIP6_HUMAN antibody
• TRIP6i2 antibody
• ZRP 1 antibody
• ZRP-1 antibody
• Zyxin-related protein 1 antibody
see all
图片
• All lanes : Anti-TRIP6 antibody (ab70747) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg
Lane 2 : HeLa whole cell lysate at 15 µg
Lane 3 : HeLa whole cell lysate at 5 µg
Lane 4 : 293T whole cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 50 kDa
Observed band size: 55 kDa (why is the actual band size different from the predicted?)
Exposure time: 3 minutes
• 1mg whole cell lysate from HeLa cells was immunprecipitated using ab70747 at 3µg/mg of lysate. For the subsequent western blot, 20% of the immunoprecipitate was probed with ab70747 at 1ug/ml. Detection: chemiluminescence with exposure time of 10 seconds.
文献
ab70747 has not yet been referenced specifically in any publications.
客户评价及客户问答
There are currently no Customer reviews or Questions for ab70747.
Please use the links above to contact us or submit feedback about this product.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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-7,184,778,478,993,925,000 | Skip to main content
Cambridge University Libraries are providing a blend of online and controlled in-person services. Please see our website for more details.
Birkbeck College, London
Organization
Found in 2 Collections and/or Records:
File
Correspondence with Birkbeck College, London, 1989 - 1993
Reference Code: GBR/0014/YUNG 6/5
Scope and Contents
Michael Young was a student on the Philosophy MA course at Birkbeck, 1989-90.
Dates: 1989 - 1993
Fonds
The Papers of Rosalind Franklin
Reference Code: GBR/0014/FRKN
Scope and Contents A lot of the material within the collection relates to Rosalind Franklin's work on the structure of deoxyribonucleic acid (DNA). Consisting of working papers, notebooks, reports and correspondence, this material is of considerable historical interest to those studying the discovery of the structure of DNA. There is also an extensive collection of papers relating to Franklin's work on the Tobacco Mosaic Virus (TMV), which took place at Birkbeck College in London. There are few papers...
Dates: 1927 - 2017
Conditions Governing Access: The collection is open for consultation by researchers using Churchill Archives Centre, Churchill College, Cambridge. Rosalind Franklin's personal correspondence (reference: FRKN 9) is closed until 2034.
Additional filters:
Type
Archival Object 1
Collection 1
Subject
Biology 1
Crystallography 1
Laboratories 1
Molecular biology 1
Philosophy 1
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3,521,344,208,207,475,000 | Full Record Display for the EPA National Library Catalog
RECORD NUMBER: 261 OF 955
OLS Field Name OLS Field Data
Main Title Evaluation of low order stream quality in central Iowa /
Author Arthur, J. W. ; Roush, T. ; Thompson, J. A. ; Walbridge, C. T. ; Puglisi, F. A.
Other Authors
Author Title of a Work
Arthur, John W.
CORP Author National Health and Environmental Effects Research Lab., Duluth, MN. Mid-Continent Ecology Div.
Publisher Mid-Continent Ecology Division, National Health and Environmental Effects Research Laboratory] ; [Distributed by National Technical Information Service],
Year Published 1997
Report Number EPA/600/R-97/009
Stock Number PB97-196398
OCLC Number 38742667
Subjects Stream ecology--Iowa.
Additional Subjects Aquatic habitats ; Biological communities ; Watersheds ; Toxicity ; Bottom sediments ; Fish populations ; Invertebrates ; Water chemistry ; Water quality ; Aquatic plants ; Drainage ; Physical properties ; Sampling ; Bioassay ; Correlation ; Land use ; Agricultural runoff ; Central Region ; Environmental stress
Internet Access
Description Access URL
http://nepis.epa.gov/Exe/ZyPDF.cgi?Dockey=30003J3R.PDF
Holdings
Library Call Number Additional Info Location Last
Modified
Checkout
Status
EHAD EPA/600-R-97-009 Region 1 Library/Boston,MA 05/08/1998
EHBD EPA/600/R-97/009 NHEERL/AED Library/Narragansett,RI 03/02/2007
EJAD EPA 600/R-97-009 Region 3 Library/Philadelphia, PA 03/28/1998
EJBD ARCHIVE EPA 600-R-97-009 Headquarters Library/Washington,DC 05/07/2013
EJBD EPA 600-R-97-009 c.1 Headquarters Library/Washington,DC 05/07/2013
EKAM 600/R-97/009 Region 4 Library/Atlanta,GA 06/16/2000
EKCD EPA/600/R-97/009 NHEERL/GED Library/Gulf Breeze,FL 10/30/2018
ELBD EPA 600-R-97-009 AWBERC Library/Cincinnati,OH 04/25/1998
ELDD EPA/600/R-97/009 3 copies NHEERL/MED Library/Duluth,MN 07/31/1998
ENAD EPA 600-R-97-009 Region 7 IRC Library/Kansas City,KS 03/22/2017
ESAD EPA 600-R-97-009 Region 10 Library/Seattle,WA 03/23/2010
NTIS PB97-196398 Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy. NTIS 01/01/1988
Collation ix, 36 pages : illustrations ; 28 cm
Abstract This research describes procedures and results obtained to evaluate the baseline (existing) watershed quality in the low order streams in a tri-county area in central Iowa. The five streams evaluated were located in the Upper Skunk River Basin. Field work was conducted over a three-year period from 1992 to 1994, and sampling conducted at 12 locations. The field procedures used physical (habitat), chemical (surface and sediment pore water quality), toxicological (daphid and algal bioassays), and biological (macroinvertebrates and fish) techniques. Habitat quality was the highest in the larger drainages. Non-farm streamside vegetative buffers were greater at the larger drainage sites. Significant associations were found among the macroinvertebrate community indices, surface and sediment pore water quality and drainage area. Correlations were also found between habitat quality and the biological community indices. Based on our measurements, lowest watershed quality was present in the upper drainage reaches. This study found that elevated concentrations of sediments and nutrients were associated with degraded biological communities found in low order agricultural streams.
Notes "PB97-196398." "EPA/600/R-97/009." "June 1997."
Place Published Duluth, MN Springfield, Va.
Supplementary Notes See also PB94-155645.
Availability Notes Order this product from NTIS by: phone at 1-800-553-NTIS (U.S. customers); (703)605-6000 (other countries); fax at (703)321-8547; and email at orders@ntis.fedworld.gov. NTIS is located at 5285 Port Royal Road, Springfield, VA, 22161, USA.
Corporate Au Added Ent National Health and Environmental Effects Research Laboratory (U.S.). Mid-Continent Ecology Division.
PUB Date Free Form 1997
NTIS Prices PC A04/MF A01
BIB Level m
Medium unmediated
Content text
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Language eng
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7,519,486,389,635,710,000 | Photo reduction in photosynthesis what is oxidized
The reduction and oxidation reactions are shortly known as redox reactions. Photosynthesis is a process in which plants containing chlorophyll convert the This generates the proton gradient which helps in the synthesis of chemical. Photosynthesis is a redox process where oxidation and reduction both occurs. CO2 + H2O Image courtesy Photosynthesis Notes. k Views · View 6. Photosynthesis is the process used by plants and a few microorganisms to transform sunlight, carbon dioxide and water into two products;.
The photo part of photosynthesis involves the oxidation of oxygen, specifically the oxygen from the water. The photo part provides us with reducing power (in the. Photosynthesis involves oxidation and reduction by oxidizing the oxygen in The photo part of photosynthesis involves the oxidation of the. Photosynthesis converts light energy into the chemical energy of sugars and other This electron transfer is an example of an oxidation-reduction process: the.
In the first step, the oxygen in water is oxidized by the light energy: . BChl a of the special pair has been photo-oxidized while the BPheo a molecule is reduced. An example of naturally-occuring biological oxidation-reduction reactions is the which may be utilized for energy as well as the synthesis of other compounds. Photosynthesis can be defined as the light-driven synthesis of carbohydrate. How can you tell if a molecule has been oxidized or reduced? (1) look for a. | {
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-8,668,400,602,434,067,000 | Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.299109
Title: Structural and functional studies on the B cell surface molecule CD22.
Author: Piperi, Christina.
ISNI: 0000 0001 3491 7579
Awarding Body: University of Oxford
Current Institution: University of Oxford
Date of Award: 1999
Availability of Full Text:
Full text unavailable from EThOS. Please contact the current institution’s library for further details.
Abstract:
No abstract available
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.299109 DOI: Not available
Keywords: Immunoglobin Biochemistry Molecular biology Cytology Genetics
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-352,292,046,817,301,300 | Register or Login
All
• All
• Uniprot Id
• Catalog #
• Peptide Sequence
> home > Products > Primary Antibodies > Signal Transduction > PLC gamma 2 Antibody
PLC gamma 2 Antibody
Purified Rabbit Polyclonal Antibody
• WB - PLC gamma 2 Antibody ABV11547-30
• SPECIFICATION
• CITATIONS
• PROTOCOLS
• BACKGROUND
Product Information
Application
• Applications Legend:
• WB=Western Blot
• IHC=Immunohistochemistry
• IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
• IHC-F=Immunohistochemistry (Frozen Sections)
• IF=Immunofluorescence
• FC=Flow Cytopmetry
• IC=Immunochemistry
• ICC=Immunocytochemistry
• E=ELISA
• IP=Immunoprecipitation
• DB=Dot Blot
• CHIP=Chromatin Immunoprecipitation
• FA=Fluorescence Assay
• IEM=Immunoelectronmicroscopy
• EIA=Enzyme Immunoassay
WB
Primary Accession P16885
Reactivity Human
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 147870 Da
Additional Information
Gene ID 5336
Other Names PLC, EC 3.1.4.11 , Phosphoinositide phospholipase C , PLC-gamma-2 , Phospholipase C-gamma-2 , PLC-IV , 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2
Target/Specificity PLC gamma 2
Formulation 100 µg (0.5 mg/ml) protein A affinity purified rabbit polyclonal antibody in phosphate-buffered saline (PBS) containing 30% glycerol, 0.5% BSA, and 0.01% thimerosal.
Handling The antibody solution should be gently mixed before use.
Background Descriptions
PrecautionsPLC gamma 2 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name PLCG2
Function The production of the second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) is mediated by activated phosphatidylinositol-specific phospholipase C enzymes. It is a crucial enzyme in transmembrane signaling.
Research Areas
Citations (0)
Thousands of laboratories across the world have published research that depended on the performance of antibodies from Abgent to advance their research. Check out links to articles that cite our products in major peer-reviewed journals, organized by research category.
Submit your citation using an Abgent antibody to
info@abgent.com, and receive a free "I Love Antibodies" mug.
Background
PLC (Phosphoinositide-specific phospholipase C) plays a significant role in transmembrane signaling. Four members of PLCs have been identified: PLCβ, PLCg, PLCδ, and PLCe. In response to extracellular stimuli (e.g., hormone, growth factors, neurotransmitters), PLC hydrolizes phosphatidylinositol 4,5-biphosphate (PIP2) into two secondary messengers: inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG). PLCg2 is engaged in antigen-dependent signaling in B-cells and collagen-dependent signaling in platelets.
FeedBack
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If you have used an Abgent product and would like to share how it has performed, please click on the "Submit Review" button and provide the requested information. Our staff will examine and post your review and contact you if needed.
If you have any additional inquiries please email technical services at tech@abgent.com.
$ 89.00
$ 245.00
Cat# ABV11547-30
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-2,798,545,898,001,733,000 | 中国农业科学院蔬菜花卉研究所2018年度公开招聘工作人员推迟报名截止时间的公告 中国农业科学院蔬菜花卉所2018年度公开招聘工作人员公告
Dissecting the genome of the polyploid crop oilseed rape by transcriptome sequencing.
来源： 发布时间：07-06 作者： 点击量：
Bancroft, I., Morgan, C., Fraser, F., Higgins, J., Wells, R., Clissold, L., Baker, D., Long, Y., Meng, J., Xiaowu Wang, Liu, S., and Trick, M.
Nature Biotechnology,29,762-766,(2011)
DOI: 10.1038/nbt.1926
Abstract: Genetic linkage maps play a key role in marker-assisted predictive crop breeding by defining the genomic positions of trait-controlling loci. Relationships between the positions of such loci and genome sequences can be deduced, at least in principle, when sequence-based molecular markers are used to construct linkage maps. This enables the identification of candidate genes linked to phenotypes of interest. For plants with relatively simple genomes, high-throughput sequencing approaches permit the assembly of sequence scaffolds representing most of the total gene space, particularly when combined with conventional Sanger sequencing2, 3, 4, 5, 6, 7. However, genomes assembled by these methods are not as complete as the earliest genomes for model species8, 9. Moreover, the capacities of whole-genome approaches are particularly limited for species such as bread wheat and oilseed rape with recent polyploidy events in their ancestry, owing to the difficulty in correctly assembling very closely related sequences.
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5,756,571,750,003,977,000 | sirt6
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• Official Full Name
• sirtuin 6
• Background
• The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as Class III histone deacetylases. The first discovered and best ch
• Synonyms
• SIRT6; sirtuin 6; sirtuin (silent mating type information regulation 2 homolog) 6 (S. cerevisiae) , sirtuin (silent mating type information regulation 2, S. cerevisiae, homolog) 6; NAD-dependent deacetylase sirtuin-6; sirtuin type 6; SIR2-like protein 6
Cat.#:SIRT6-213HTag:T7/His
Source (Host):E. coliSpecies:Human
Product nameRecombinant Human SIRT6 protein, T7/His-tagged
+Inquiry
Cat.#:SIRT6-2684HTag:GST
Source (Host):E. coliSpecies:Human
Product nameRecombinant Human SIRT6, GST-tagged
+Inquiry
Cat.#:SIRT6-28517THTag:T7
Source (Host):E. coliSpecies:Human
Product nameRecombinant Human SIRT6, T7 -tagged
+Inquiry
Cat.#:SIRT6-357HTag:His
Source (Host):E. coliSpecies:Human
Product nameRecombinant Human Sirtuin 6, His-tagged
+Inquiry
Cat.#:SIRT6-4882HTag:His
Source (Host):Sf 9 Insect cellsSpecies:Human
Product nameRecombinant Human Sirtuin 6, His-tagged
+Inquiry
Cat.#:SIRT6-4883HTag:GST
Source (Host):E. coliSpecies:Human
Product nameRecombinant Human Sirtuin 6, GST-tagged
+Inquiry
Cat.#:Sirt6-1381MTag:His
Source (Host):E. coliSpecies:Mouse
Product nameRecombinant Mouse Sirt6 protein, His-tagged
+Inquiry
Cat.#:Sirt6-1382RTag:His
Source (Host):E. coliSpecies:Rat
Product nameRecombinant Rat Sirt6 protein, His-tagged
+Inquiry
Cat.#:SIRT6-3378CTag:His
Source (Host):Mammalian CellsSpecies:Chicken
Product nameRecombinant Chicken SIRT6
+Inquiry
Cat.#:SIRT6-304ZTag:His
Source (Host):Mammalian CellsSpecies:Zebrafish
Product nameRecombinant Zebrafish SIRT6
+Inquiry
Cat.#:SIRT6-1829HCLTag:
Source (Host):Species:Human
Product nameRecombinant Human SIRT6 293 Cell Lysate
+Inquiry
Cat.#:Kit-2341Tag:
Source (Host):Species:
Product nameSIRT6 Inhibitor Screening Kit (Fluorometric)
+Inquiry
Involved Pathway
SIRT6 involved in several pathways and played different roles in them. We selected most pathways SIRT6 participated on our site, such as Central carbon metabolism in cancer, which may be useful for your reference. Also, other proteins which involved in the same pathway with SIRT6 were listed below. Creative BioMart supplied nearly all the proteins listed, you can search them on our site.
Pathway Name
Pathway Related Protein
Central carbon metabolism in cancer
KIT; MAPK3; HK1; PDGFRA; PIK3CA; SCO2; NTRK3; RET; PIK3R3; TRP53
Protein Function
SIRT6 has several biochemical functions, for example, NAD(P)+-protein-arginine ADP-ribosyltransferase activity, NAD+ ADP-ribosyltransferase activity, NAD+ binding. Some of the functions are cooperated with other proteins, some of the functions could acted by SIRT6 itself. We selected most functions SIRT6 had, and list some proteins which have the same functions with SIRT6. You can find most of the proteins on our site.
Function
Related Protein
Function NAD(P)+-protein-arginine ADP-ribosyltransferase activity
Related Protein SIRT6; CHAT1; Art2b; ART1; ART3; MADPRT; ART5; CHAT2
Function NAD+ ADP-ribosyltransferase activity
Related Protein SIRT4; TNKS; PARP12A; GIG2H; PARP11; GIG2G; GIG2Q; PARP1; PARP15; GIG2O
Function NAD+ binding
Related Protein ALDH1A3; SIRT7; SIRT3; HADH; SIRT6; HPGD; SIRT5; SIRT2; SIRT4; GLUD1
Function NAD-dependent histone deacetylase activity
Related Protein SIRT2; SIRT6; SIRT1
Function NAD-dependent histone deacetylase activity (H3-K9 specific)
Related Protein SIRT1; SIRT6
Function chromatin binding
Related Protein MEN1; HIF1AB; KAT2B; PRKCBB; RNF20; CENPB; ANKRD17; RNF168; ARID3C; SKIL
Function protein binding
Related Protein IFI16; SLC30A3; HDAC7A; PHF8; ANGPTL4; RFC5; CCL5; FOXR2; SLA2; SCUBE3
Function transcription corepressor activity
Related Protein TLE4; LHX9; SKOR1A; SMYD1; SSX5; TRIB3; HOMEZA; ID4; MXD4; SNW1
Function zinc ion binding
Related Protein SUV39H2; NR2F1; USP51; CPO; PRKCBA; Car8; TRHDE; LMO2; NR2E1; MME
Interacting Protein
SIRT6 has direct interactions with proteins and molecules. Those interactions were detected by several methods such as yeast two hybrid, co-IP, pull-down and so on. We selected proteins and molecules interacted with SIRT6 here. Most of them are supplied by our site. Hope this information will be useful for your research of SIRT6.
RELA; VIM; CHD3; MYC; a8k1f4_human; MLF1; HSF2; PSMD2; q96be0_human; AARSD1; FKBPL; STUB1; UBE2D1
SIRT6 Related Articles
Tahir, MN; Ragg, R; et al. Amine functionalized ZrO2 nanoparticles as biocompatible and luminescent probes for ligand specific cellular imaging. JOURNAL OF MATERIALS CHEMISTRY B 3:2371-2377(2015).
Orellana, ME; Quezada, C; et al. Expression of SIRT2 and SIRT6 in Retinoblastoma. OPHTHALMIC RESEARCH 53:100-108(2015). | {
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5,327,649,520,338,874,000 | Submit Data | Help | Video Tutorials | News | Publications | FTP Download | REST API | Citing RGD | Contact
ONTOLOGY REPORT - ANNOTATIONS
Term:positive regulation of T cell tolerance induction
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Accession:GO:0002666 term browser browse the term
Definition:Any process that activates or increases the frequency, rate, or extent of T cell tolerance induction.
Synonyms:exact_synonym: positive regulation of T lymphocyte tolerance induction; up regulation of T cell tolerance induction; upregulation of T cell tolerance induction
narrow_synonym: activation of T cell tolerance induction; stimulation of T cell tolerance induction
show annotations for term's descendants Sort by:
positive regulation of T cell tolerance induction term browser
Symbol Object Name Evidence Notes Source PubMed Reference(s) RGD Reference(s) Position
G FOXP3 forkhead box P3 ISO (MGI:3692513|PMID:16945588) MGI PMID:16945588 MGI:3692513 NCBI chr X:43,303,777...43,328,164
Ensembl chr X:43,303,782...43,328,735
JBrowse link
G IDO1 indoleamine 2,3-dioxygenase 1 IEA Ensembl GO_REF:0000107 NCBI chr17:9,245,283...9,260,140
Ensembl chr17:9,245,196...9,260,491
JBrowse link
G TGFBR2 transforming growth factor beta receptor 2 IEA Ensembl GO_REF:0000107 NCBI chr13:16,784,370...16,875,828
Ensembl chr13:16,784,491...16,878,165
JBrowse link
positive regulation of peripheral T cell tolerance induction term browser
Symbol Object Name Evidence Notes Source PubMed Reference(s) RGD Reference(s) Position
G FOXP3 forkhead box P3 IEA Ensembl GO_REF:0000107 NCBI chr X:43,303,777...43,328,164
Ensembl chr X:43,303,782...43,328,735
JBrowse link
positive regulation of T cell anergy term browser
Symbol Object Name Evidence Notes Source PubMed Reference(s) RGD Reference(s) Position
G CBLB Cbl proto-oncogene B ISO (MGI:3038342|PMID:14973438) MGI PMID:14973438 MGI:3038342 NCBI chr13:153,480,525...153,700,115
Ensembl chr13:153,395,978...153,700,064
JBrowse link
G CD3E CD3e molecule ISO (MGI:3038342|PMID:14973438) MGI PMID:14973438 MGI:3038342 NCBI chr 9:45,619,730...45,633,972
Ensembl chr 9:45,619,481...45,635,379
JBrowse link
G FOXP3 forkhead box P3 IEA Ensembl GO_REF:0000107 NCBI chr X:43,303,777...43,328,164
Ensembl chr X:43,303,782...43,328,735
JBrowse link
G ITCH itchy E3 ubiquitin protein ligase IEA Ensembl GO_REF:0000107 NCBI chr17:37,810,113...37,918,963
Ensembl chr17:37,810,183...37,918,208
JBrowse link
G LOC100517285 leukocyte immunoglobulin-like receptor subfamily A member 6 ISO (PMID:16493035) ARUK-UCL PMID:16493035 NCBI chr 6:58,996,449...59,001,088 JBrowse link
Term paths to the root
Path 1
Term Annotations click to browse term
biological_process 16548
immune system process 2302
tolerance induction 31
T cell tolerance induction 14
positive regulation of T cell tolerance induction 7
positive regulation of T cell anergy 5
positive regulation of peripheral T cell tolerance induction + 1
Path 2
Term Annotations click to browse term
biological_process 16548
developmental process 6302
anatomical structure development 5842
multicellular organism development 5340
system development 4963
immune system development 1030
tolerance induction 31
T cell tolerance induction 14
regulation of T cell tolerance induction 12
positive regulation of T cell tolerance induction 7
positive regulation of T cell anergy 5
positive regulation of peripheral T cell tolerance induction + 1
paths to the root
NHLBI Logo
RGD is funded by grant HL64541 from the National Heart, Lung, and Blood Institute on behalf of the NIH. | {
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-8,880,042,661,828,485,000 |
Department of Energy Argonne National Laboratory Office of Science NEWTON's Homepage NEWTON's Homepage
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NEWTON Home Page NEWTON Teachers Visit Our Archives Ask A Question How To Ask A Question Question of the Week Our Expert Scientists Volunteer at NEWTON! Frequently Asked Questions Referencing NEWTON About NEWTON About Ask A Scientist Education At Argonne Gene Mutations
Name: Jessica
Status: Educator
Grade: 6-8
Location: CA
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Date: November 2005
Question:
The question is regarding mutation of genes. The students want to know when and where on the gene does mutation occur?
Replies:
A mutation is a change in the base sequence of DNA...it was originally defined for those protein coding parts of the DNA (gene)...as opposed to the stretches of DNA that do not code for protein, but the more encompassing definition is any change in the base sequence of DNA...With this in hand, a mutation can occur anywhere on the chromosomal DNA. When, is another matter. Mutations can occur anytime...during replication DNA polymerase can make a mistake...not often. It can also occur following exposure to radiation. UV light causes adjacent thymine bases (on the same strand)to pair up...called thymine dimmers.
pf
Mutations are any change in the DNA sequence. It can be as simple as a one letter change, or as much as a whole part of a chromosome being lost, which changes many genes at once. Mutations aren't always a bad thing-sometimes the mutation occurs in part of the gene so that the protein's shape doesn't change enough to effect its function. Other times, the one letter changes the shape so drastically that it can't function at all. So it's hard to say-there isn't one answer to your question. But very basically-the DNA codes for amino acids which are in turn part of a protein. The protein has a shape, let's say like a key. Also, for that protein to work properly that shape needs to fit with something else, say like a lock. If the key changes shape it won't fit in its lock any more and can't do its job. But if the change is in the part of the key that doesn't actually fit in the lock, it won't be as bad.
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5,275,145,004,457,593,000 |
Motor neuron-derived Thsd7a is essential for zebrafish vascular development via the Notch-dll4 signaling pathway
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Motor neuron-derived Thsd7a is essential for zebrafish vascular development via the Notch-dll4 signaling pathway
Post Jon Moulton on Tue Aug 16, 2016 3:50 pm
Liu LY, Lin MH, Lai ZY, Jiang JP, Huang YC, Jao LE, Chuang YJ. Motor neuron-derived Thsd7a is essential for zebrafish vascular development via the Notch-dll4 signaling pathway. J Biomed Sci. 2016 Aug 2;23(1):59. doi: 10.1186/s12929-016-0277-9.
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-7,221,596,578,897,527,000 | Skip to main content
Figure 3 | BMC Bioinformatics
Figure 3
From: A robust and efficient algorithm for the shape description of protein structures and its application in predicting ligand binding sites
Figure 3
Definition of true/false positives and true/false negatives for the predicted ligand binding site residues evaluated with respect to the referenced ligand binding site in a protein. True and false positives are the correctly and incorrectly predicted number of binding site residues in a protein, respectively. True and false negatives are the correctly and incorrectly predicted number of non-binding site residues, respectively. They are defined for each known ligand binding site on a protein by protein basis.
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1 October 2001
75. Sterol 14α-Demethylase as a Potential Target for Antitrypanosomal Therapy: Enzyme Inhibition and Parasite Cell Growth
Galina I. Lepesheva, Robert D. Ott, Tatiana Y. Hargrove, Yuliya Y. Kleshchenko, Inge Schuster, W. David Nes, George C. Hill, Fernando Villalta, Michael R. Waterman
Summary | | | Supplemental Data
Chemistry & Biology, 14(11)
26 November 2007
76. Cellular Energy Depletion Resets Whole-Body Energy by Promoting Coactivator-Mediated Dietary Fuel Absorption
Atul R. Chopra, Ramakrishna Kommagani, Pradip Saha, Jean-Francois Louet, Christina Salazar, Junghun Song, Jaewook Jeong, Milton Finegold, Benoit Viollet, Franco DeMayo et al.
Summary | | | Supplemental Data
Cell Metabolism, 13(1)
5 January 2011
77. Determination of the Orientation and Dynamics of Ergosterol in Model Membranes Using Uniform 13C Labeling and Dynamically Averaged 13C Chemical Shift Anisotropies as Experimental Restraints
O. Soubias, F. Jolibois, S. Massou, A. Milon, V. Réat
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Biophysical Journal, 89(2)
1 August 2005
78. Membrane rafts in plant cells
Sébastien Mongrand, Thomas Stanislas, Emmanuelle M.F. Bayer, Jeannine Lherminier, Françoise Simon-Plas
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Trends in Plant Science, 15(12)
1 December 2010
79. Ablation of gp78 in Liver Improves Hyperlipidemia and Insulin Resistance by Inhibiting SREBP to Decrease Lipid Biosynthesis
Tong-Fei Liu, Jing-Jie Tang, Pei-Shan Li, Yang Shen, Jia-Gui Li, Hong-Hua Miao, Bo-Liang Li, Bao-Liang Song
Summary | | | Supplemental Data
Cell Metabolism, 16(2)
8 August 2012
80. Common Single-Nucleotide Polymorphisms Act in Concert to Affect Plasma Levels of High-Density Lipoprotein Cholesterol
Victor Spirin, Steffen Schmidt, Alexander Pertsemlidis, Richard S. Cooper, Jonathan C. Cohen, Shamil R. Sunyaev
Abstract | |
The American Journal of Human Genetics, 81(6)
1 December 2007
81. Brassinosteroid biosynthesis inhibitors
Tadao Asami, Shigeo Yoshida
Abstract | |
Trends in Plant Science, 4(9)
1 September 1999
82. Visualization of Lipid Metabolism in the Zebrafish Intestine Reveals a Relationship between NPC1L1- Mediated Cholesterol Uptake and Dietary Fatty Acid
James W. Walters, Jennifer L. Anderson, Robert Bittman, Michael Pack, Steven A. Farber
Summary | | | Supplemental Data
Chemistry & Biology, 19(7)
27 July 2012
83. Joe Goldstein and Mike Brown: from cholesterol homeostasis to new paradigms in membrane biology
Richard G.W Anderson
Abstract | |
Trends in Cell Biology, 13(10)
1 October 2003
84. Potato glycoalkaloids: true safety or false sense of security?
Yaroslav I. Korpan, Elena A. Nazarenko, Irina V. Skryshevskaya, Claude Martelet, Nicole Jaffrezic-Renault, Anna V. El'skaya
Abstract | |
Trends in Biotechnology, 22(3)
1 March 2004
85. Hepatic Stearoyl-CoA Desaturase-1 Deficiency Protects Mice from Carbohydrate-Induced Adiposity and Hepatic Steatosis
Makoto Miyazaki, Matthew T. Flowers, Harini Sampath, Kiki Chu, Carolin Otzelberger, Xueqing Liu, James M. Ntambi
Summary | | | Supplemental Data
Cell Metabolism, 6(6)
5 December 2007
86. Brassinosteroid Signal Transduction
Steven D Clouse
Summary | |
Molecular Cell, 10(5)
1 November 2002
87. Molecular Mechanisms of Hepatic Steatosis and Insulin Resistance in the AGPAT2-Deficient Mouse Model of Congenital Generalized Lipodystrophy
Víctor A. Cortés, David E. Curtis, Suja Sukumaran, Xinli Shao, Vinay Parameswara, Shirya Rashid, Amy R. Smith, Jimin Ren, Victoria Esser, Robert E. Hammer et al.
Summary | | | Supplemental Data
Cell Metabolism, 9(2)
4 February 2009
88. LXRs regulate the balance between fat storage and oxidation
Nada Y. Kalaany, Karine C. Gauthier, Ann Marie Zavacki, Pradeep P.A. Mammen, Tatsuya Kitazume, Julian A. Peterson, Jay D. Horton, Daniel J. Garry, Antonio C. Bianco, David J. Mangelsdorf
Summary | | | Supplemental Data
Cell Metabolism, 1(4)
1 April 2005
89. A New Role for a Metabolic Star: AMP-Activated Protein Kinase Stimulates Fat Absorption
Pascal Ferré, Fabienne Foufelle
Summary | |
Cell Metabolism, 13(1)
5 January 2011
90. Plant ABC proteins – a unified nomenclature and updated inventory
Paul J. Verrier, David Bird, Bo Burla, Elie Dassa, Cyrille Forestier, Markus Geisler, Markus Klein, Üner Kolukisaoglu, Youngsook Lee, Enrico Martinoia et al.
Abstract | | | Supplemental Data
Trends in Plant Science, 13(4)
1 April 2008
91. Vitamins in plants: occurrence, biosynthesis and antioxidant function
M. Amparo Asensi-Fabado, Sergi Munné-Bosch
Abstract | |
Trends in Plant Science, 15(10)
1 October 2010
92. Impaired Generation of 12-Hydroxylated Bile Acids Links Hepatic Insulin Signaling with Dyslipidemia
Rebecca A. Haeusler, Matthew Pratt-Hyatt, Carrie L. Welch, Curtis D. Klaassen, Domenico Accili
Summary | | | Supplemental Data
Cell Metabolism, 15(1)
4 January 2012
93. Nowa1p and Nowa2p: Novel Putative RNA Binding Proteins Involved in trans-Nuclear Crosstalk in Paramecium tetraurelia
Mariusz Nowacki, Wlodzimierz Zagorski-Ostoja, Eric Meyer
Summary | | | Supplemental Data
Current Biology, 15(18)
20 September 2005
94. Minimal Effect of Lipid Charge on Membrane Miscibility Phase Behavior in Three Ternary Systems
Matthew C. Blosser, Jordan B. Starr, Cameron W. Turtle, Jake Ashcraft, Sarah L. Keller
Abstract | | | Supporting Material
Biophysical Journal, 104(12)
18 June 2013
95. Engineering flax and hemp for an alternative to cotton
Michel J.M Ebskamp
Abstract | |
Trends in Biotechnology, 20(6)
1 June 2002
96. FXR: a promising target for the metabolic syndrome?
Bertrand Cariou, Bart Staels
Abstract | |
Trends in Pharmacological Sciences, 28(5)
1 May 2007
97. Nocturnin Regulates Circadian Trafficking of Dietary Lipid in Intestinal Enterocytes
Nicholas Douris, Shihoko Kojima, Xiaoyue Pan, Alexandra F. Lerch-Gaggl, Son Q. Duong, M. Mahmood Hussain, Carla B. Green
Summary | | | Supplemental Data
Current Biology, 21(16)
23 August 2011
98. Ezetimibe Blocks Internalization of the NPC1L1/Cholesterol Complex
Ta-Yuan Chang, Catherine Chang
Summary | |
Cell Metabolism, 7(6)
4 June 2008
99. Apoptotic Cells Promote Their Own Clearance and Immune Tolerance through Activation of the Nuclear Receptor LXR
Noelia A-Gonzalez, Steven J. Bensinger, Cynthia Hong, Susana Beceiro, Michelle N. Bradley, Noam Zelcer, Jose Deniz, Cristina Ramirez, Mercedes Díaz, German Gallardo et al.
Summary | | | Supplemental Data
Immunity, 31(2)
21 August 2009
100. Predicting drug sensitivity and resistance
Gergely Szakács, Jean-Philippe Annereau, Samir Lababidi, Uma Shankavaram, Angela Arciello, Kimberly J. Bussey, William Reinhold, Yanping Guo, Gary D. Kruh, Mark Reimers et al.
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Cancer Cell, 6(2)
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Explore Life (with CD-ROM and InfoTrac),9780030225581
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Summary
This fresh approach to teaching introductory biology to non-science majors is based on two sets of beliefs. First, that nonscience majors absorb biology best when it is presented in the context of personal experience or human relevance. Second, that today's variety of media--video, CD-ROMs, and the Internet--offer unprecedented opportunities to bring people, imagery, biological processes, hands-on interactivity, and up-to-the-minute data into the study of biology. Authors John Postlethwait and Janet Hopson have created a topical, case-history approach to teaching life science fundamentals that fully integrates electronic media so that the student can explore relevant biology in ways suited to his or her own learning style. An engaging, student-relevant case study opens and is interwoven throughout each chapter. Additional examples, anecdotes, research stories, and applications are also used to give students a well-rounded framework for learning core biological concepts. The book's innovative design allows integration of references to the other EXPLORE LIFE components within it. The media are integrated into the course line by line, not just attractively packaged and offered as ancillaries. This complete integration can be seen in every chapter. This multimedia, applications-driven approach in EXPLORE LIFE has the potential to reshape the teaching of non-majors biology. Instructors can adopt this approach with a high level of flexibility, using as much or as little of the media as they deem appropriate, incorporating their own material, and adapting EXPLORE LIFE to their individual needs.
Table of Contents
1. What Is Life (And Why Study It)? Case Study Search for Life on Mars
Part 1. CELLS, GENES AND LIFE'S PERPETUATION
2. Cells and the Chemistry of Life
Case Study HIV and AIDS
3. How Cells Take in and Use Energy
Case Study Metabolism of Hummingbirds
4. The Cell Cycle
Case Study Cancer and the Cell Cycle
5. Patterns of Inheritance
Case Study Cystic Fibrosis
6. DNA and the Thread of Life
Case Study DNA Fingerprinting
7. Gene Function and Manipulation
Case Study Transgenic Goats
8. Reproduction and Development
Case Study In Vitro Fertilization
Part 2. EVOLUTION AND BIODIVERSITY
9. Mechanisms of Evolution
Case Study Antibiotic Resistance
10. Life's Origins and Biodiversity
Case Study Extreme Environments in Hawaii
11. Single-celled Life
Case Study Malaria (Plasmodium)
12. Fungi and Plants
Decomposers and Producers
Case Study Chile peppers
13. Animals
the Great Consumers
Case Study Dung Beetles
Part 3. THE DYNAMIC BODY
14. Body Function, Survival and Steady State
Case Study Effects of Weightlessness in Outer Space
15. Circulation and Respiration
Case Study Blood Substitutes
16. Immune System
Case Study Battling the Common Cold/The Flu
17. Nutrition and Digestion
Case Study Koala Digestion
18. Hormones
Messengers of Change
Case Study Stress Hormones in a College Student
19. Nervous System
Case Study Brain Mapping and Neural Activity in Deaf People
20. Muscles and Skeleton
The Body in Motion
Case Study Locomotion and Balance in Athletes vs
Older People
Part 4. THE LIVING PLANT
21. Plant Life
Form and Function
Case Study Hemp Anatomy and Uses
22. How Plants Grow
Case Study Genetically Engineered Rice
23. The Dynamic Plant
Case Study Weeds and Super Weeds
Part 5. THE LIVING WORLD
24. Ecology of Populations and Communities
Case Study Overfishing Off Cape Cod
25. Ecology of Ecosystems and Biosphere
Case Study Global Warming and Shifting Butterfly Ranges
26. Animal Behavior
Case Study Sex Lives of Bonobos and Chimps
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8,656,364,893,472,903,000 | Skip Navigation
Search
The Structure of Cells
The Structure of Cells
Cells are often referred to as the “building blocks of life.” All living organisms contain these smallest units of life that can replicate independently.
Each cell, whether prokaryotic or eukarotic, contains a structural, functional and biological purpose. Discover more about the structure of cells.
Author(s): Christine Herrmann, PhD
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Infectious Disease Agents
Introduction to Phylogenic Kingdoms
The Kingdom Monera - Eubacteria
The Kingdom Monera - Archaebacteria
Ecological Importance of Prokaryotes
Let's Talk About Bacteria
Structures and Functions of Genomes
The Chromosome
Prokaryotic Genomes
RNA Processing in Eukaryotic Cells | {
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-7,308,684,433,240,843,000 | Aging Cell. 2009 Dec; 8(6): 624–632.
PMCID: PMC2805862
Calorie restriction reduces rDNA recombination independently of rDNA silencing
Abstract
Calorie restriction (CR) extends lifespan in yeast, worms, flies and mammals, suggesting that it acts via a conserved mechanism. In yeast, activation of the NAD-dependent histone deacetylase, Sir2, by CR is thought to increase silencing at the ribosomal DNA, thereby reducing the recombination-induced generation of extrachromosomal rDNA circles, hence increasing replicative lifespan. Although accumulation of extrachromosomal rDNA circles is specific to yeast aging, it is thought that Sirtuin activation represents a conserved longevity mechanism through which the beneficial effects of CR are mediated in various species. We show here that growing yeast on 0.05 or 0.5% glucose (severe and moderate CR, respectively) does not increase silencing at either sub-telomeric or rDNA loci compared with standard (2% glucose) media. Furthermore, rDNA silencing was unaffected in the hxk2Δ, sch9Δ and tor1Δ genetic mimics of CR, but inhibited by FOB1 deletion. All these interventions extend lifespan in multiple yeast backgrounds, revealing a poor correlation between rDNA silencing and longevity. In contrast, CR and deletion of the FOB1, HXK2, SCH9 and TOR1 genes, all significantly reduced rDNA recombination. This silencing-independent mechanism for suppressing rDNA recombination may therefore contribute to CR-mediated lifespan extension.
Keywords: aging, dietary restriction, longevity, Sir2, telomere, yeast
Introduction
In recent years, there has been an increasing recognition that similar fundamental cellular processes underlie aging in all eukaryotes (Guarente & Kenyon, 2000). Perhaps the most persuasive evidence that aging has a common cellular basis across phyla comes from studies of dietary limitation, often termed calorie restriction (CR) (Bishop & Guarente, 2007). Although data are not available for humans, CR has been known for over 70 years to increase lifespan in rodents, with more recent studies confirming the generality of this phenomenon in both vertebrates and invertebrates (Merry, 2005; Partridge et al., 2005). Despite this, the precise molecular mechanism by which CR increases longevity in diverse species remains unclear.
Studies using the budding yeast, Saccharomyces cerevisiae, have been at the forefront of recent efforts to understand the molecular mechanism of action of CR. Yeast lifespan can be measured in two ways. Replicative lifespan is defined as the number of buds produced by an individual yeast mother cell and is independent of calendar time (Mortimer & Johnston, 1959; Muller et al., 1980); in contrast to the alternative measure of the duration of viability in stationary phase, termed chronological lifespan (MacLean et al., 2001). Reducing the concentration of glucose in yeast growth media from the standard 2% to 0.5% or below increases both replicative and chronological lifespan irrespective of genetic background and has been suggested to be a model of CR (Jiang et al., 2000; Lin et al., 2000; Fabrizio et al., 2005; Smith et al., 2007).
Although the effect of glucose limitation in extending yeast replicative lifespan is not disputed, its mechanism of action is the subject of considerable controversy and heated debate. Three main models have been suggested, two of which centre on the evolutionarily conserved NAD-dependent histone deacetylase, Sir2 (Fig. 1). Initially, it was proposed that CR causes a metabolic shift away from fermentation and towards respiration, resulting in an increased NAD:NADH ratio, and thus increased Sir2 activity (Lin et al., 2000, 2002, 2004) (model 1). Alternatively (model 2), it was suggested that CR does not alter the NAD:NADH ratio, but rather acts by increasing expression of Pnc1, which degrades the endogenous Sir2 inhibitor nicotinamide, thus increasing Sir2 activity (Anderson et al., 2003). In both models, activation of Sir2 leads to increased rDNA silencing and a consequent reduction in recombination between rDNA repeats; this reduces the formation of extrachromosomal rDNA circles (Sinclair & Guarente, 1997) and so retards aging.
Fig. 1
Proposed mechanisms of lifespan extension by calorie restriction. Models 1 and 2 suggest that CR up-regulates Sir2 activity, which leads to increased gene silencing and decreased rDNA recombination. Consequently, less ERCs are formed, which extends the ...
In contrast, the observation that CR could extend lifespan in some sir2 deletion strains (Jiang et al., 2002; Kaeberlein et al., 2004) led to the proposal of a third mechanism (Fig. 1, model 3). This postulates that CR extends lifespan in a Sir2-independent manner via inhibition of the Tor and Sch9 kinase signalling pathways (Kaeberlein et al., 2004, 2005c). In this model, the downstream molecular mechanism effecting longevity is not entirely clear, but may be related to reduced ribosomal protein biogenesis (Kaeberlein et al., 2005c; Steffen et al., 2008). Controversially, it has recently been claimed that such Sir2-independent effects of CR are in fact due to activation of Sir2 homologues and so simply result in the same increase in rDNA silencing invoked in models 1 and 2 (Lamming et al., 2005; Medvedik et al., 2007) [but see (Kaeberlein et al., 2006)].
Despite the popularity of models 1 and 2, there is surprisingly little published data demonstrating that CR does in fact increase transcriptional silencing at the rDNA locus (Lin et al., 2002). Indeed, most published work has exploited ‘genetic mimics’ of CR and assayed rDNA recombination, rather than directly analysing silencing in response to reduced glucose levels. Furthermore, it has recently been shown that severe CR has no effect on transcriptional silencing at telomeres, which is a similarly Sir2-dependent process (Kaeberlein et al., 2005a,c;). We therefore set out to determine the effect of CR and genetic mimics of CR on rDNA silencing and recombination. We found that rDNA silencing was unaffected by CR or by deletion of HXK2, SCH9 or TOR1, whereas deletion of FOB1 greatly inhibited rDNA silencing. In contrast, all of these well established lifespan-extending interventions reduced rDNA recombination. Taken together, these data are consistent with a silencing-independent mechanism for suppression of rDNA recombination in CR-mediated lifespan extension.
Results
If CR acts via Sir2 activation (Fig. 1, models 1 and 2), then this would be predicted to result in increased silencing at subtelomeric loci. However, it has recently been claimed that severe CR imposed by growth on 0.05% glucose has no effect on telomeric silencing in the PSY316 yeast strain (Kaeberlein et al., 2005c). We therefore investigated the effects of both moderate (0.5% glucose) and severe (0.05% glucose) CR on telomeric silencing using a well established reporter strain, AEY1017, which has a URA3 marker integrated into a subtelomeric region of chromosome VII (Meijsing & Ehrenhofer-Murray, 2001). Expression of the URA3 gene from nonsilenced genomic loci enables growth on media lacking uracil, but prevents growth on media containing FOA, because of conversion to the toxic product, 5-fluorouracil. However, integration of URA3 into subtelomeric regions results in partial silencing of the reporter gene and hence the unusual ability to grow both on media lacking uracil and on media containing FOA. Deletion of the SIR2 gene inhibits telomeric silencing, resulting in increased expression of the URA3 reporter gene, which is routinely visualized as decreased growth on FOA. Indeed, no growth was observed on FOA plates for the sir2 deletion strain (Fig. 2A), thus confirming the validity of this approach for assaying in vivo Sir2 activity. In contrast to the profound effect of sir2 deletion on silencing, no obvious difference in growth on FOA plates was seen between 2%, 0.5% and 0.05% glucose (Fig. 2A).
Fig. 2
Calorie restriction does not affect subtelomeric gene silencing. (A) Qualitative silencing assay. The telomeric silencing reporter parent strain, AEY1017, and its isogenic sir2Δ strain were grown up in YPD, then diluted in H2O and serially spotted ...
Although CR had no obvious effect on telomeric silencing in spot tests, subtle effects could be missed in this simple visual assay. We therefore performed quantitative silencing assays by counting the number of individual colonies formed on selective FOA plates relative to the total number of colonies on nonselective media, as previously described (Kaeberlein et al., 2005c). Increased silencing should result in increased viability in this assay, but over a series of 6 independent experiments totalling more than 60 000 colonies counted, we found no significant difference between 2%, 0.5% and 0.05% glucose (Fig. 2B). Thus, these quantitative silencing assays confirmed the findings from spot test assays: neither moderate nor severe CR affects telomeric silencing.
Although CR had no effect on telomeric silencing, it nevertheless remained possible that rDNA silencing was indeed increased. We therefore utilized the widely used rDNA reporter strain, JS128, which has a URA3 marker integrated into the nontranscribed spacer region 1 of the rDNA (Smith & Boeke, 1997). Again, partial silencing of this reporter gene enables growth on media lacking uracil and media containing FOA. Deletion of the SIR2 gene inhibits rDNA silencing, resulting in increased expression of the URA3 reporter gene, which is routinely visualized in such rDNA reporter strains as increased growth on −ura plates (Smith & Boeke, 1997; Smith et al., 2007) (Fig. 3A). If CR increases rDNA silencing, this should be seen as decreased growth on −ura plates relative to nonselective media. However, no gross effects were seen in spot tests using 0.5% and 0.05% glucose (Fig. 3A). To quantify these effects, a similar approach was taken to that used for the telomeric reporter assays described above, where the number of viable colonies on −ura plates relative to nonselective media was calculated. Over a series of three independent experiments with over 50 000 colonies counted, no significant difference in rDNA silencing was seen between 2% and 0.5% glucose (Fig. 3B). However, a partial reduction in viability on −ura plates was noted at 0.05% glucose. Such reduced growth on −ura plates could either be due to increased rDNA silencing or to a synthetic growth phenotype caused by the simultaneous imposition of the double stress of limiting glucose and uracil levels. To control for the latter, duplicate assays were performed in parallel using a different selective media, FOA, where increased silencing could not correlate with decreased colony growth (Fig. 3B, grey bars). This revealed a similar growth inhibition at 0.05% glucose (but not at 0.5% glucose), indicating that the partial reduction in viability on −ura media observed is not due to increased rDNA silencing. Confirmation of the artefactual nature of this reduced growth at 0.05% glucose was evident by comparison with a positive control sir2Δ strain (Fig. 3B), which exhibited the expected reciprocal relationship between growth on the two media (increased on −ura, decreased on FOA). Thus, both spot test and quantitative silencing assays indicate that moderate and severe CR have no effect on rDNA silencing.
Fig. 3
Calorie restriction does not affect gene silencing in the rDNA. (A) Qualitative silencing assay. The rDNA silencing reporter parent strain, JS128, and its isogenic sir2Δ strain were grown up in YPD, then diluted in H2O and serially spotted out ...
The correlation between increased rDNA silencing and longevity is integral to Sirtuin-dependent models of yeast lifespan extension. As we found no effect of CR on rDNA silencing, we investigated whether deletion mutations that reproducibly extend lifespan in multiple yeast background strains alter rDNA silencing. Deletion of HXK2, one of three hexokinases in yeast, limits the entry of glucose into glycolysis and hence is widely used as a genetic mimic of CR acting upstream of Sir2/Tor/Sch9. Indeed, like CR, HXK2 deletion has been shown to increase lifespan in multiple yeast genetic backgrounds (Lin et al., 2000; Kaeberlein et al., 2005b). Deletion of FOB1 is thought to increase lifespan by reducing rDNA recombination [and hence extrachromosomal rDNA circle (ERC) generation] downstream of Sir2 (Defossez et al., 1999; Kaeberlein et al., 2005b). In Fig. 4A, it can be seen that the hxk2Δ strain behaves similarly to the JS128 parent strain on all media, indicating no major effect on rDNA silencing. However, the fob1Δ strain exhibited enhanced growth on −ura plates, and decreased growth on FOA plates, similar to the sir2Δ strain, indicating a loss of rDNA silencing. In contrast, deletion of RPD3, which is known to increase rDNA silencing in a Sir2-dependent manner (Sun & Hampsey, 1999), gave the expected decrease in growth on −ura plates, thus confirming that this assay is able to detect such increases in rDNA silencing. To check if these effects were rDNA-specific, we deleted FOB1 and HXK2 in a control strain (JS122) where the URA3 reporter gene is integrated in a nonsilenced genomic locus. Here, there was no difference in growth on selective media between the parent strain and its isogenic mutants: all grew similarly on −ura plates, none grew on FOA (Fig. 4B). Finally, we investigated whether telomeric silencing was similarly affected by creating deletion mutants in the AEY1017 strain. In spot test assays, the hxk2Δ strain was similar to its isogenic parent strain and to a fob1Δ strain (Fig. 4C). Therefore, the hxk2Δ genetic mimic of CR, like CR itself, has little effect on silencing at rDNA or telomeric loci, whereas deletion of FOB1 specifically inhibits rDNA silencing.
Fig. 4
Effects of lifespan-extending mutations on rDNA silencing. (A) The rDNA silencing reporter parent strain, JS128, and isogenic deletion strains hxk2Δ and fob1Δ, were grown up in YPD. These were then diluted in H2O and serially spotted out ...
Despite these different effects on silencing, both CR and deletion of FOB1 have been reported to extend lifespan via inhibition of rDNA recombination. We therefore assayed mitotic rDNA recombination by monitoring the frequency of URA3 marker loss after serial culturing for approximately 120 generations (Dror & Winston, 2004). Over a series of experiments, the mean frequency of marker loss in the parent JS128 strain at 2% glucose was 1.3 × 10−3 per generation, similar to previously reported values for other strains (Merker & Klein, 2002; Lamming et al., 2005). Deletion of FOB1 reduced the frequency of URA3 marker loss by around 90%, whereas deletion of SIR2 caused an approximately threefold increase in marker loss (Fig. 5A), consistent with the known pro- and anti-recombinase functions of Fob1 and Sir2, respectively, at the rDNA (Merker & Klein, 2002). These effects on URA3 marker loss were specific for the rDNA locus, as no marker loss was observed over 120 generations in the JS122 parent strain where URA3 is integrated outside the rDNA, or in its isogenic JS122 fob1Δ and sir2Δ strains (Fig. 5A). These data confirmed the validity of assaying the frequency of URA3 marker loss in the JS128 strain as a readout of rDNA recombination rate, thus enabling the effects of CR to be determined. Both moderate (0.5% glucose) and severe (0.05% glucose) CR produced a significant reduction in rDNA recombination of around 20% (Fig. 5B). Therefore, both CR and FOB1 deletion act via a silencing-independent mechanism to reduce rDNA recombination.
Fig. 5
Calorie restriction reduces rDNA recombination. The rDNA silencing reporter parent strain, JS128, the control JS122 strain, and isogenic fob1Δ and sir2Δ strains were serially cultured in YP media containing the indicated glucose concentrations ...
Finally, we investigated whether the Sir2-independent Sch9/Tor1-mediated longevity pathway of model 3 might also impact on rDNA silencing and recombination. Deletion of SCH9 appeared to slightly improve growth on −ura media relative to nonselective media in qualitative spot test assays (Fig. 6A), similar to the general CR mimic hxk2Δ (Fig. 4A, ,6A);6A); whereas, deletion of TOR1 appeared to slightly decrease growth on −ura plates (Fig. 6A). Although these small effects were also apparent in quantitative silencing assays (Fig. 6B), the differences in viability on −ura plates between the various deletion mutants and the isogenic parent strain were not statistically significant. Therefore, like CR itself, genetic mimics of CR do not cause significant increases in rDNA silencing. In contrast, the hxk2Δ, sch9Δ and tor1Δ strains all exhibited significantly reduced rDNA recombination (Fig. 6C), reinforcing the notion that various lifespan-extending interventions can reduce rDNA recombination independently of rDNA silencing.
Fig. 6
Genetic mimics of calorie restriction reduce rDNA recombination independently of rDNA silencing. (A) Qualitative silencing assay. The rDNA silencing reporter parent strain, JS128, and its isogenic sch9Δ, tor1Δ and sir2Δ strains ...
Discussion
Reducing the concentration of glucose from the standard 2% to 0.5% or below has been reported to extend replicative lifespan in all yeast strains examined (Jiang et al., 2000; Lin et al., 2000; Kaeberlein et al., 2004; Lamming et al., 2005). Likewise, deletion of SIR2 shortens replicative lifespan and overexpression of Sir2 extends replicative lifespan in most strains (Kaeberlein et al., 1999, 2004; Kim et al., 1999; Anderson et al., 2003). While there is little doubt that both CR and Sir2 are powerful and near-universal modulators of longevity, debate has raged over whether the two are part of a single mechanism or represent separate pathways (Kaeberlein & Powers, 2007). Sir2 is an attractive candidate for a central mediator of CR, as its NAD-dependent histone deacetylase activity could potentially sense CR-induced increases in the NAD:NADH ratio or decreases in nicotinamide levels and transduce these into increased rDNA silencing and consequently reduced rDNA recombination. The reported lack of effect of severe CR on telomeric silencing (Kaeberlein et al., 2005c), which we have confirmed here and extended to moderate CR, argues against this idea, as does the observation that CR can extend lifespan in some SIR2 deletion strains (Jiang et al., 2002; Kaeberlein et al., 2004). Nevertheless, this does not rule out the possibility that CR acts specifically on the rDNA to increase silencing. Indeed, a recent screen for genes that increase rDNA silencing in the absence of Sir2 has claimed that Sir2-independent lifespan extension by CR in fact proceeds via activation of the Sir2 homologue, Hst2, and so ultimately works by the same mechanism of increased rDNA silencing/decreased rDNA recombination (Lamming et al., 2005). However, the data presented here strongly suggest that CR has no effect on rDNA silencing, and thus acts via a pathway(s) that does not involve activation of Sir2 or its homologues.
How can we reconcile our data with earlier work that established models 1 and 2? While there is no doubt that Sir2 histone deacetylase activity is of primary importance in establishing rDNA silencing, a survey of the literature reveals surprisingly little data directly demonstrating that CR affects rDNA silencing. Indeed, to our knowledge, the only such data is Fig. 1d in (Lin et al., 2002), where the result of a qualitative assay using MET15 as a reporter gene inserted in the rDNA was presented. The increased brown colouration of this strain on 0.5% glucose compared with 2% glucose was consistent with an effect of moderate CR increasing rDNA silencing. However, such assays are not quantifiable and are subject to interpretations over ‘shades of brownness’. Furthermore, similar effects of CR on MET15 reporter gene expression are observed when the marker is integrated into nonsilenced regions of the genome, suggesting that the increased pigmentation observed is a metabolic effect unrelated to silencing (Smith et al., 2009). The data presented here, using both qualitative and quantitative assays of URA3 reporter gene expression, strongly argue that CR has no effect on rDNA silencing. As CR has no effect on telomeric silencing either (this study and Kaeberlein et al., 2005c), it seems unlikely that activation of Sir2 (or its homologues) occurs in response to CR. Furthermore, rDNA silencing is not significantly affected in the CR genetic mimics, hxk2Δ, sch9Δ and tor1Δ; whereas fob1Δ strains actually exhibit greatly reduced rDNA silencing. As CR and all these deletion mutants have been shown to increase lifespan in various genetic backgrounds, these observations further emphasize the lack of correlation between rDNA silencing and lifespan, and so make it unlikely that Sirtuin-mediated rDNA silencing is crucial for longevity.
What implications do our data have for the competing theories of CR outlined in Fig. 1? It is important to note that although our data clearly argue against the essential role for Sir2 activation and increased rDNA silencing implicit in models 1 and 2, they are nevertheless consistent with the proposal that CR extends lifespan by reducing rDNA recombination, albeit via a silencing-independent mechanism. Indeed, the observation that FOB1 deletion inhibits rDNA silencing (this study and Huang & Moazed, 2003), clearly demonstrates that these two processes can be uncoupled; as FOB1 deletion strongly decreases rDNA recombination (this study and Defossez et al., 1999; Kobayashi et al., 1998). As Fob1 is a component of the RENT complex that targets Sir2 to the nucleolus (Huang & Moazed, 2003), the consequent loss of Sir2 from the rDNA explains the reduced silencing in FOB1 deletion strains, but this in turn inevitably suggests that the decrease in rDNA recombination is Sir2- and silencing-independent. It has previously been shown that rDNA recombination is reduced by moderate CR, rapamycin and by the putative CR genetic mimic cdc25-10 (Lin et al., 2000; Lamming et al., 2005; Medvedik et al., 2007). We report here that both moderate and severe CR, as well as deletion of the HXK2, SCH9 and TOR1 genes all reduce rDNA recombination. Thus, it may be that models 1 and 2 are essentially correct in that CR ultimately extends lifespan, at least in part, by decreasing rDNA recombination, but that this occurs by an (unknown) Sirtuin-independent process. In both models, the longevity associated with reduced rDNA recombination is attributed to reduced ERC generation. While this remains a plausible mechanism, it is interesting to note that in fob1/sir2 double mutants, which have extremely low ERC levels, CR increases lifespan and reduces rDNA recombination (Kaeberlein et al., 1999, 2004; Lamming et al., 2005). In addition, lifespan reduction is associated with increased rDNA recombination in the face of unchanged ERC levels in sgs1, hpr1 and dna2 mutants (Heo et al., 1999; McVey et al., 2001; Hoopes et al., 2002; Mankouri et al., 2002; Merker & Klein, 2002). Further work is required to determine if the CR-induced reduction in rDNA recombination impacts on longevity via ERCs or a different mechanism.
Although the Sir2-independent, Sch9/Tor1-regulated model 3 centres upon reduced ribosomal protein synthesis as the downstream mechanism of lifespan extension (Kaeberlein et al., 2005c; Steffen et al., 2008), our data clearly show that these interventions also reduce rDNA recombination. Therefore, it is possible that CR and all its various genetic mimics extend lifespan, at least in part, by a common action in reducing rDNA recombination. The molecular mechanisms involved in this putative silencing-independent regulation of rDNA recombination are unknown. However, it is interesting to note that Tor1 physically interacts with the 35S rDNA promoter (Tsang et al., 2003) and that its association with the rDNA is regulated by nutrient levels (Li et al., 2006). In addition, it has recently been shown that localization of rDNA repeats to the inner nuclear membrane suppresses rDNA recombination independently of rDNA silencing (Mekhail et al., 2008). Clearly, dissection of the molecular mechanisms involved in yeast lifespan extension by CR will require much further work in order to construct a single unifying model.
Experimental procedures
Chemicals and reagents
Materials for yeast culture were obtained from Sigma-Aldrich (Poole, UK) and Foremedium (Norwich, UK). 5-Fluoro-orotic acid (FOA) was obtained from Apollo Scientific (Stockport, UK). PCR primers were supplied by Sigma Genosys (Havenhill, UK), genomic DNA isolation kits were from Invitrogen (Paisley, UK); and PCR enzymes/reagents were from Promega (Southampton, UK). All other materials were obtained from Sigma-Aldrich.
Strain construction
Telomeric silencing in this study was assessed using the telomeric URA3 marker strain, AEY1017 (W303-1B; Matαade2-1 ura3-1 his3-11,15 leu2-3, 112 trp1-1 can 1-100 TELVIIL::URA3) (Meijsing & Ehrenhofer-Murray, 2001). Silencing at rDNA was assessed using the rDNA URA3 marker strain, JS128(S6) (JB740; MATαhis3Δ200 leu2Δ1 ura3-167 RDN1::Ty1::mURA3) (Smith & Boeke, 1997). As a control for silencing-independent effects, a strain with URA3 integrated at a nonsilenced locus was used, JS122 (JB740; MATαhis3Δ200 leu2Δ1 ura3-167 ??::Ty1::mURA3) (Smith & Boeke, 1997). Deletion strains with the appropriate open reading frame replaced by the KanMX4 cassette in the BY4741 MATa haploid background were obtained from Invitrogen. Deletion cassettes for FOB1, HXK2, SIR2 and TOR1 were PCR amplified from genomic DNA prepared from the respective BY4741 deletion strain. The SCH9 deletion cassette was PCR amplified from the pFA6KanMX4 plasmid (Longtine et al., 1998) using appropriate primers. The resulting PCR products were used to transform each URA3 reporter strain via the PCR-based disruption strategy (Schiestl & Gietz, 1989; Wach, 1996). Successful G418-resistant transformants were confirmed by PCR using gene-specific and KanMX primers (data not shown). Strains used in this study are listed in Table 1.
Table 1
Yeast strains used in this study
Qualitative silencing assays (spottests)
For silencing assays, synthetic complete media (SC), uracil omission media (SC − ura) and FOA-supplemented media (SC + FOA) were prepared from 2× stock solutions and D-glucose added to 0.05%, 0.5% and 2% from a 50% stock solution. 5-Fluoro-orotic acid was prepared in DMSO and added to media to give a final concentration of 1 mg ml−1 and 1% DMSO final concentration. Single colonies were picked from a plate and grown overnight in 5 mL liquid YP media containing 2% glucose. The next morning, cultures were diluted to OD600 = 1 in sterile H2O in 1 mL. The adjusted cultures were serially diluted five times in sterile H2O at a ratio of 1:5 starting from OD600 = 1 in column 1 in a 96 well-plate to 100 μL final volume and plated with a replica plater onto SC, SC − ura and SC + FOA plates containing 0.05%, 0.5% and 2%, glucose and incubated at 30°C for 2 (0.5% and 2% glucose) to 4 days (0.05% glucose). Plates were imaged in a BioRad Universal Hood II Imager (BioRad, Hemel Hempstead, UK).
Quantitative silencing assays
Cultures were grown overnight in 5 mL YPD, diluted with water to OD600 = 1 in 1 mL final volume. For silencing assays using the parent strains, cultures were then serially diluted in sterile H2O to 10−3, 5 × 10−3, 10−4, 5 × 10−4, and 100 μL of each dilution was plated on SC, SC − ura, or SC + FOA plates supplemented with the various glucose concentrations. The plates were incubated at 30°C for 2–4 days. For silencing assays using deletion strains, overnight liquid cultures were adjusted to OD600 = 1 in 1 mL, and serially diluted. On SC and SC − ura plates, 100 μL of a 10−4 dilution were plated, and on SC + FOA, 100 μL of a 10−3 dilution were plated. Colonies were counted either manually or with a BioRad Universal Hood II Imager and QuantityOne software (Biorad). Viability of the AEY1017 telomeric reporter strain on FOA medium is directly proportional to silencing activity (Meijsing & Ehrenhofer-Murray, 2001). Viability was calculated by dividing the number of colonies on FOA plates either by the number of colonies on SC plates (FOA/SC) or by the total number of colonies on −ura and FOA plates combined (FOA/−ura + FOA), both of which yielded similar results. Data from assays of six independently grown yeast clones were pooled and statistical analysis performed using Student’s t-tests. Viability of the JS128 rDNA reporter strain on −ura medium is inversely proportional to silencing activity (Smith & Boeke, 1997) and was calculated by dividing the number of colonies on −ura plates by the number of colonies on SC plates (-ura/SC). To control for synthetic growth effects of selective media at low glucose concentrations, duplicate assays were performed in parallel on FOA plates and viability calculated by dividing the number of colonies on FOA plates by the number of colonies on SC plates. Four platings per strain/condition were analysed for each rDNA silencing assay, and every assay was performed on at least two independently grown yeast clones. The resulting data were then pooled and statistical analysis performed using Student’s t-tests.
rDNA recombination assays
Mitotic stability of the URA3 gene in JS122 and JS128 strains was assayed using the method of Dror and Winston, 2004. Briefly, single colonies from -ura plates were grown to saturation in 10 mL YPD at 30 °C. At this point, the cultures were diluted 1:10 000 in 10 mL fresh YPD and grown to saturation once more. After nine such saturation passages (equivalent to approximately 120 generations), each culture was spread onto ten YPD plates at an appropriate dilution to yield 50–300 colonies/plate. After growth at 30 °C for 2–3 days, each plate was then replica plated onto both SC and −ura plates and grown for several days at 30 °C. Frequency of URA3 marker loss by recombination was calculated by dividing the number of colonies that did not grow on −ura plates by the total number of colonies on replica SC or YPD plates. Marker loss per generation was calculated by dividing this value by the total number of mitotic divisions during the nine serial passages (Cole et al., 2007). Ten platings per strain/condition were used for each recombination assay, and every assay was performed on at least two independently grown yeast clones. The resulting data were then pooled and statistical analysis performed using Student’s t-tests.
Acknowledgments
The authors thank Dr Jeff Smith (University of Virginia, USA) for the gift of the JS122 and JS128 strains, and for communicating unpublished results. The authors thank Drs Jessica Downs and Bessie Bilsland (University of Sussex, UK) for the gift of the AEY1017 strain and for advice and assistance with silencing assays. This work was supported by the Wellcome Trust in the form of a Prize Studentship to MR.
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NB11058358
0.1 ml
Each for $458.00
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Description
Description
YAP1 Polyclonal antibody specifically detects YAP1 in Human, Mouse, Drosophila samples. It is validated for Western Blot, Simple Western, Chromatin Immunoprecipitation, Immunohistochemistry, Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunoblotting, Knockout Validated.
Specifications
Specifications
Rabbit
This YAP1 Antibody was developed against a partial recombinant human YAP1 protein expressed in bacteria. [Uniprot: P46937], N-terminal GST fusion protein
Polyclonal
Primary
Western Blot, ChIP assay, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunohistochemistry (Frozen), Immunoblot, Immunoassay, KnockDown
P46937
PBS, 15% glycerol with 0.02% Sodium Azide
IgG
YAP1
Affinity Purified
RUO
Expected reactivity based on immunogen homology: Isoform 4 (100%), Isoform 6 (100%)
10413
0.2 mg/mL
SDS
Product Certifications
For Research Use Only
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4,809,947,796,588,571,000 |
Overview
• Product name
Anti-pCrosstide antibody (HRP)
• Description
Rabbit polyclonal to pCrosstide (HRP)
• Host species
Rabbit
• Conjugation
HRP
• Specificity
Recognizes pCrosstide only. Does not cross-react with unphosphorylated Crosstide.
• Tested applications
Suitable for: WB, ELISAmore details
• Species reactivity
Reacts with: Species independent
• Immunogen
pCrosstide peptide conjugated to KLH.
Properties
Applications
Our Abpromise guarantee covers the use of ab21626 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes
WB Use a concentration of 0.5 µg/ml.
ELISA Use a concentration of 0.25 µg/ml.
Target
• Relevance
Crosstide is a synthetic peptide derived from the N terminal sequence of GSK3 and is a well documented substrate of PKB (AKT) and SGK kinases. pCrosstide is the phosphorylated product of these kinases.
• Alternative names
• PRAApTF antibody
References
ab21626 has not yet been referenced specifically in any publications.
Customer reviews and Q&As
There are currently no Customer reviews or Questions for ab21626.
Please use the links above to contact us or submit feedback about this product.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Sign up | {
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-6,976,125,355,154,865,000 | 24 visiting now: 18 USA USA flag 2 China China flag 2 France France flag 1 Germany Germany flag 1 Pakistan Pakistan flag
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-7,178,667,576,353,716,000 | FIND THE ANSWERS
What happends when you move vegetation from land?
Answer this question
• What happends when you move vegetation from land?
Answers
What happends when you move vegetation from land? resources. Land versus Water Heating and Cooling ... Love Happens / New York, I Love You / Land of the ...
Read more
Positive: 51 %
... a nine-acre plot of densely wooded land has been sealed off from the ... are constantly on the move. ... and surrounding vegetation, ...
Read more
Positive: 48 %
More resources
Explain how energy is transferred through food chains and ... % of THAT energy is passed onto them so as you can see the further down the food chain ...
Read more
Positive: 51 %
This Is What Happens to Your Body After You Die. Moheb ... “They move into the ... it kills off some of the underlying and surrounding vegetation, ...
Read more
Positive: 46 %
... while starlings can detect insecticidal compounds in vegetation, ... If you suspect that a rabbit's nest has been abandoned, ...
Read more
Positive: 32 %
There have been many civilian casualties resulting from the land mines. ... on the steppes is mostly semi-desert vegetation, ... Nagorno-Karabakh Republic:
Read more
Positive: 9 %
Show more results | {
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-8,263,556,784,606,709,000 | Welcome Guest, please Log in shopping cartYour cart is empty
KOMP Repository (Knockout Mouse Project) KOMP Knock Out Knockout Mouse Mice
St6galnac5
ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5
Synonyms: Siat7e, ST6GalNAc V
Gene nomenclature, locus information, and GO, OMIM, and PMID associations are updated daily from MGI
Gene Identification
Targeting Projects
Available
Click to order
Not Currently Available
Click to see options for obtaining this product.
Project ID Allele Product Availability
Vectors ES Cells Mice Germplasm
Mice Cryo
Recovery
Microinject Sperm Embryos
CSD83920 St6galnac5tm1a(EUCOMM)Wtsi
Knockout First
(Promoter driven)
St6galnac5tm1a
Knockout First
(Promoter driven)
VG14869 St6galnac5tm1(KOMP)Vlcg
Deletion
See Details
CSD28048 St6galnac5
CSD28049 St6galnac5
Non-KOMP projects associated with this gene exist at: EUCOMM. You can view the status of all IKMC projects for St6galnac5.
Researchers interested in St6galnac5 are also interested* in
Aldoa Aspa Agtr2 Alas2 Acta2 Adrbk2 Ambp Ace Acvr1 Adra2a
* Based on co-mentions in published literature. For more gene-gene associations visit genecloud.org.
The KOMP Repository is located at the University of California Davis and Children’s Hospital Oakland Research Institute. Question? Comments? For Mice, Cells, and germplasm please contact us at service@komp.org, US 1-888-KOMP-MICE or International +1-530-752-KOMP, or for vectors komporders@chori.org or +1-510-450-7917. | {
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-7,563,454,593,944,498,000 | Navigation Links
Rapid evolution within single crop-growing season increases insect pest numbers
Date:8/15/2011
populations.
"This shows that even without human interference natural selection acting on aphid populations causes rapid evolution," said Martin M. Turcotte, who led the research as a graduate student in ecology, evolution and organismal biology at UC Riverside. "Even stronger effects might be expected when pesticides are in use. For decades, evolution was deemed too slow and, hence, it was not considered when studying population growth an oversight that needs to be corrected. Ignoring this evolution, as is not currently uncommon, can lead to predictions that greatly underestimate pest densities and outbreaks."
Rapid evolution could have important untested impacts in many other applied areas. For example, rapid evolution is important in fisheries where intense fishing causes fish to evolve traits that let them escape fishing nets. Antibiotic resistance and increased virulence in pathogens are examples where rapid evolution impacts human health.
The study was conducted at the University of California Motte Rimrock Reserve where the researchers collected multiple clonal lineages from a wild aphid population feeding on mustard plants. They identified clones and characterized their intrinsic per capita growth rates in a greenhouse at UCR.
'/>"/>
Contact: Iqbal Pittalwala
iqbal@ucr.edu
951-827-6050
University of California - Riverside
Source:Eurekalert
Page: 1 2
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Rapid evolution within single crop-growing season increases insect pest numbers
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990,852,689,648,466,400 | discontinued
14-9930-81 H-Y TCR (male antigen HY) antibody
See related secondary antibodies
Search for all "H-Y TCR (male antigen HY)"
50 µg / €200.00
Quick Overview
Mouse anti Human H-Y TCR (male antigen HY) T3.70
14-9930-81
Product Description for H-Y TCR (male antigen HY)
Mouse anti Human H-Y TCR (male antigen HY) T3.70.
Presentation: Purified
Product is tested for Flow Cytometry.
Properties for H-Y TCR (male antigen HY)
Product Category Primary Antibodies
Quantity 50 µg
Presentation Purified
Reactivity Hu
Applications F
Clonality Monoclonal
Clone T3.70
Host Mouse
Isotype IgG1
Shipping to Germany only
PDF datasheet View Datasheet
Manufacturer eBioscience Inc.
Datasheet Extract
Isotype control AM03095PU-N, SM10P (for use in human samples)
Format
State:
Purified
Accessory Products
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Article
A Litopenaeus vannamei Hemocyanin-Derived Antimicrobial Peptide (Peptide B11) Attenuates Cancer Cells’ Proliferation
1
Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China
2
STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou 515063, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Molecules 2018, 23(12), 3202; https://doi.org/10.3390/molecules23123202
Received: 26 October 2018 / Revised: 15 November 2018 / Accepted: 2 December 2018 / Published: 5 December 2018
(This article belongs to the Collection Natural Products: Anticancer Potential and Beyond)
Abstract
:
Antimicrobial peptides play important roles in the immune response to pathogens and tumor cells; for this reason, they are being exploited for therapeutic use. In this study, we describe a Litopenaeus vannamei hemocyanin-derived peptide, denoted B11, which shares similar features with other anticancer peptides and attenuates the proliferation of cancer cells. Cell viability assay revealed that B11 significantly inhibited the proliferation of human cervical (HeLa), human hepatocellular carcinoma (HepG2), and human esophageal cancer (EC109) cancer cell lines, but not normal liver cell lines (T-antigen-immortalized human liver epithelial (THLE) cells or THLE-3), by inducing morphological changes, nuclear condensation, and margination, features which are indicative of apoptosis. Besides, peptide B11-induced apoptosis was confirmed by isothiocyanate-labeled Annexin V/propidium iodide (Annexin V-FITC/PI) double staining of HeLa cells. Moreover, cell uptake studies, confocal microscopy, and Western blot analysis revealed that rhodamine-labeled B11 permeated HeLa cells and localized to the mitochondria, causing mitochondria dysfunction through lost mitochondrial membrane potential, which consequently triggered the induction of apoptosis. Increased expression levels of caspase-9, caspase-3, and Bax (Bcl-2-associated X) proteins, coupled with a decrease in Bcl-2 (B-cell lymphoma 2) protein, confirmed that peptide B11 induced apoptosis via the mitochondrial pathway. Thus, the hemocyanin-derived peptide, B11, inhibits the proliferation of cancer cells by causing mitochondrial dysfunction and inducing apoptotic cell death, for which reason it could be explored as an anticancer peptide.
1. Introduction
Hemocyanin (HMC) is a respiratory protein, which is also reported to be multi-functional, and thus plays essential roles in mollusk and arthropods [1,2]. A growing number of immune-related functions have been ascribed to hemocyanin [3,4], including phenoloxidase [5], antiviral [6], agglutinative [7,8], anticancer [9], reaction with anti-human Ig (immunoglobulin) as an antigen [10], hemolysin activity [11], and as an immune-enhancement protein in shrimp [12]. Moreover, in different crustaceans, hemocyanin is reported to generate antimicrobial peptides (AMPs) in response to microbial challenge [13]. Destoumieux-Garzón et al. reported that the C-terminal fragment of hemocyanin from penaeid shrimps Penaeus vannamei and Penaeus stylirostris had broad antifungal activities [14]. An antibacterial peptide with 16 amino acid residues was also found in the plasma of freshwater crayfish Pacifastacus leniusculus [15]. Similarly, we previously found an 18.4-kDa fragment of hemocyanin with antimicrobial activity in Litopeneus vannamei infected with Vibrio parahaemolyticus [16]. Generally, AMPs are small cationic peptides characterized by positive charges and hydrophobic amino acids, as well as amphipathic features [17]. Since AMPs are positively charged, they are able to bind to negatively charged bacteria cell membranes, resulting in the disruption of the membrane and bacteria death [18]. These features and properties of AMPs makes them important components of the innate immune system in a variety of organisms, including plants and animals [19,20].
Several recent studies have shown that AMPs also have anticancer activity [21]. For instance, Rodrigues et al. reported that a cream that is mixed with the AMP gomesin, and used as a topical drug to smear over the external surface of tumors, successfully treated intradermal and intraepithelial cancers [22]. A synthetic 21-mer AMP (Epinecidin-1) from grouper (Epinephelus coioides) has been reported to have in vitro antitumor activity against human fibrosarcoma cells (HT1080) [23]. Similarly, the pleurocidin family of AMPs (NRC-03 and NRC-07) derived from Atlantic flounder were found to kill breast carcinoma cells, including drug-resistant and slow-growing breast cancer cells [24]. Interestingly, our recent studies involving the screening of L. vannamei hemocyanin identified 20 potential AMPs ranging from 1.5 to 1.9 kDa [25]. While the antibacterial activities of these hemocyanin-derived peptides have been ascertained, whether or not these peptides also have anticancer effects is not known.
In the current study, we report on the antiproliferative and potential anticancer activity of one of these L. vannamei hemocyanin-derived AMPs (designated B11). Peptide B11 could inhibit the proliferation of three cancer cell lines by permeabilizing, entering, and inducing apoptotic cell death. Given the properties exhibited by peptide B1, it could be used for anticancer agents, while the knowledge gained from this study could provide the basis for developing therapeutic peptides from marine resources into anticancer therapeutic agents.
2. Results
2.1. Synthesis and Characterization of Peptides
The L. vannamei hemocyanin-derived antimicrobial peptide (B11) was synthesized manually via solid phase peptide synthesis (SPPS) using the Fluorenylmethyloxycarbonyl/tert-butyloxycarbonyl (Fmoc/tBu) strategy [26]. The purity and molecular mass of the successfully purified peptide B11 using the SPPS method was ascertained by reverse phase high performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis (Figure 1).
2.2. Effect of Peptide B11 on Cancer Cells’ Proliferation
The antiproliferative activity of peptide B11 against some cancer cell lines, including HeLa cells (human cervical cancer cells), HepG2 cells (human hepatocellular carcinoma cells), and EC109 cells (human esophageal cancer cells) was examined. When the cell proliferation or viability following treatment with peptide B11, 5-fluorouracil (5-FU), or PBS (Phosphate-buffered saline) was determined using the MTS assay (Figure 2), it was observed that the proliferation of all three cancer cell types was significantly decreased 24 h post-treatment with peptide B11 or with the anticancer drug 5-FU compared with PBS. For instance, peptide B11 significantly (p < 0.05) decreased HeLa cells viability by 20.0% (Figure 2A), while that of HepG2 cells decreased by 23.0% (Figure 2B) and EC109 cells decreased by 13.0% (Figure 2C) relative to PBS treatment. On the contrary, peptide B11 had no significant effect on the proliferation of normal liver cell lines (THLE-3) (Figure 2D), as treatment with B11 or PBS for 24 h had almost the same cell viability (97.92% and 100%, respectively). Thus, these results suggest that peptide B11 selectively inhibits the in vitro proliferation of only cancer cell lines, and could potentially be used as an antitumor agent.
2.3. Peptide B11 Induces Apoptosis in HeLa Cells
The induction of apoptosis induction and changes to cell morphology were examined to ascertain if this could be responsible for the antiproliferative effect of peptide B11. As shown in Figure 3A, HeLa cells treated with peptide B11 (50 μg/mL) for 24 h were changed morphologically, including cell shrinkage and rounding up. A similar observation was found in 5-FU-treated cells, but not in the PBS-treated cells. In addition, the typical characteristics of apoptosis, including nuclei shrinkage, condensation, margination and the formation of apoptotic body-like vesicles were observed in cells 24 h post-treatment with peptide B11 and 5-FU, but not in PBS (Figure 3B). Moreover, flow cytometry analysis with Annexin V/propidium iodide (Annexin V/PI) staining revealed that the ratio of viable cells to total cells gradually decreased in peptide B11-treated cells, while there was a time-dependent gradual increase in the percentage of late apoptotic cells (Figure 3C). In fact, the average percentage of apoptotic cells in peptide B11-treated cells was 40.4% at 48 h post-treatment, which is similar to that observed in the 5-FU treated cells (41.1%) at the same time point. These results suggest that peptide B11 exerts a proapoptotic effect on cancer cells, which is synonymous with 5-FU, and that this might account for the antiproliferative effects of peptide B11 on HeLa cells.
2.4. Physicochemical Properties of B11 and Cell Permeabilization Analysis
Peptide B11 is located within the copper-containing domain of the large molecular weight L. vannamei hemocyanin protein (GeneBank accession No.: CAB85965.1), and is composed of 15 amino acids, half of which are hydrophobic (Figure 4A). The physicochemical properties of peptide B11, including molecular weight, total hydrophobic ratio, hydrophobicity, hydrophobic moment, average hydropathy value, molar extinction coefficient, charge and isoelectric point are summarized in Table 1. Peptide B11 has a molecular weight of 1767 Da, with a net positive charge, and a total hydrophobic ratio of 46%. Modeling using the Schiffer–Edmundson helical wheel on the DNAstar (Lasergene 7.1, Madison, WI, USA) program revealed that peptide B11 has an amphipathic α-helical conformation (Figure 4B), while three-dimensional (3D) model prediction using the I-TASSER program (http://zhanglab.ccmb.med.umich.edu/I-TASSER) showed that it has α-helical and one β-sheet structure (Figure 4C). Thus, given the size of peptide B11, its net positive charge, α-helix, and β-sheet structure, as well as other features that are typical of therapeutic peptides and/or AMPs [27,28], it should be able to permeable cells [29].
Cell uptake studies were carried out to determine whether peptide B11 could indeed penetrate cell membranes. Confocal microscopy analysis showed that rhodamine-labeled B11 was readily taken up by HeLa cells, with the number of fluorescent cells higher at 24 h post-treatment compared to 8 h, suggesting that peptide B11 penetrates cells time-dependently (Figure 4D(i)). Moreover, analysis of the X–Z and Y–Z sections of the three-dimensional (3D) reconstructed confocal images confirmed that peptide B11 was found inside the cytoplasm of cells (Figure 4D(ii)), further suggesting that as a cationic peptide, B11 could permeate cells to exert its effects.
2.5. Peptide B11 Induces Mitochondrial-Dependent Apoptotic Cell Death in HeLa Cells through Lost of Mitochondrial Membrane Potential
The integrity of mitochondrial function is crucial for the maintenance of cell viability [30]. Thus, given that peptide B11 has a net positive charge, induces apoptosis, and could permeate cells, we went about determining the effect of peptide B11 on the mitochondria. First, confocal microscopy revealed that rhodamine-labeled B11 colocalized with MitoTracker in the mitochondria of HeLa cells (Figure 5A). Moreover, peptide B11 not only localized to the mitochondria, but was able to cause mitochondrial dysfunction in the form of a loss in mitochondrial membrane potential (Figure 5B). In addition, Western blot analysis was used to further examine the expression of proteins involved in the mitochondrial-dependent apoptosis pathways following treatment with peptide B11. As shown in Figure 5C(i), at 24-h post-treatment of HeLa cells with B11 (50 μg/mL), there was an increase in the protein expression levels of caspase-3 (an executioner caspase in the apoptosis cascade) and caspase-9 (an initiator caspase in the mitochondrial-dependent or intrinsic apoptosis pathway) compared to PBS treatment. Besides, there was an increase in the protein expression level of proapototic Bax (Figure 5C(ii)) and a decrease in the protein expression level of prosurvival Bcl-2 (Figure 5C(iii)), two Bcl-2 family members that play a central role in regulating changes in mitochondrial outer membrane permeability [31,32]. All of these results indicate that peptide B11 was able to cause mitochondrial dysfunction and induced apoptosis via the mitochondrial-dependent pathway.
3. Discussion
Several recent studies have reported on the development and sophisticated resistance being mounted by cancer cells and bacteria against most of the traditional drugs [33,34,35]. Therefore, researchers are exploring alternative more efficacious drugs, with more attention being drawn toward the use of naturally occurring antimicrobial peptides (AMPs). Several AMPs have so far been evaluated in both preclinical and clinical studies [36,37], with data suggesting that AMPs could substitute traditional antibiotics in combatting microbial infections, especially multidrug-resistant microbes [17,38]. Most importantly, some AMPs have been shown to have antitumor activity [39,40]. In the current study, an in silico predicted AMP (designated peptide B11) derived from the hemocyanin protein of the shrimp L. vannamei, was synthesized manually via solid phase peptide synthesis, and shown to significantly inhibit the proliferation of cancer cells. The antiproliferative effect of peptide B11 is because it causes mitochondrial dysfunction and induces apoptosis, therefore suggesting its anticancer potential. The research approach that was used in this study implies that bioinformatics prediction tools could be leveraged to explore anticancer therapeutic agents followed by cell-based validation.
While a number of studies have explored the discovery of AMPs, and studied their structures and mechanisms of action [41,42], recent reviews have reported on the anticancer activities and efficacy of AMPs from terrestrial animals and plants [21,43,44,45]. The shrimp L. vannamei is reported to generate different AMPs from hemocyanin, both naturally as part of the innate immune system and in response to specific pathogen challenge [13,14,46]. The antimicrobial activities of these shrimp hemocyanin-derived AMPs have been demonstrated [25]; however, their effect on cell proliferation and potential use as antiproliferative or antitumor agents has not been explored.
In this study, the antitumor potential of a 15 amino acids hydrophobic cationic antimicrobial peptide, peptide B11, which is derived from the large molecular weight hemocyanin protein of L. vannamei, and has α-helical and β-sheet structure, was explored. The structure and physicochemical properties of AMPs are reported to play a significant role in their ability to induce apoptosis in cancer cells [21,43]. Cationic AMPs are able to physically associate with the negatively charged membranes of cancer cells, destabilizing the lipid membrane and subsequently binding to intracellular targets, resulting in cell death [24]. On the other hand, AMPs rich in hydrophobic amino acids are able to insert into cell membranes to form a stable structure, thereby disrupting the cell membrane and forming pores, leading to changes in cell membrane charge and therefore interfering with cell death pathways [21]. Given that the features of peptide B11 (Table 1 and Figure 4A) are synonymous with cationic anticancer peptides, this suggests that peptide B11 might share similar properties and/or functions. Since the outer membranes of cancer cells are negatively-charged due to an abundance of anionic molecules, while the membranes of noncancerous or normal cells are neutral [44,47], cationic peptides such as B11 could exert antiproliferative effects targeted at only cancer cells, due to this membrane charge difference. In the current study, rhodamine-labeled B11 was able to permeate HeLa cells (Figure 4D), just as other anticancer peptides [40,48], and localized to the mitochondria (Figure 5A), where it induced a loss of mitochondrial membrane potential (Figure 5B), and consequently mitochondrial dysfunction. Dysfunctional mitochondria might eventually lead to the induction of apoptosis via the mitochondrial-dependent pathway [49].
In the apoptosis pathway, caspases (cysteine proteases) act in concert in a cascade to trigger apoptosis [50,51]. The apoptosis-related caspases are generally divided into initiator caspases (including caspase-2, caspase-8, caspase-9, and caspase-10), and effector or executioner caspases (including caspase-3, caspase-6, and caspase-7) [51,52,53]. As the initiator caspase in the mitochondrial-dependent apoptotic pathway [54], activated caspase-9 in turn activates caspase-3/7 potentiating the apoptosis cascade, and thereby catalyzing the cleavage of many key cellular proteins that commit cells to apoptosis. To ascertain that there was an induction of apoptosis due to the mitochondrial dysfunction, an increase in the protein expression levels of caspase-9 and caspase-3 (determined by Western blot) was observed following the treatment of HeLa cells with peptide B11 (Figure 5C(i)). Since the Bcl-2 family proteins, which govern mitochondrial outer membrane permeability, are either proapoptotic (Bax, BAD, Bak, Bok, etc.) or antiapoptotic (Bcl-2, Bcl-xL, Bcl-w, etc.) [55], we went on to determine changes in the protein expression of Bcl-2 and Bax, given their important role in mitochondrial-dependent apoptosis [31,56]. Interestingly, peptide B11 treatment caused a decrease in antiapoptotic Bcl-2 protein, but an increase in proapototic Bax in HeLa cells (Figure 5C(ii),(iii)). This observation is similar to that previously reported by Wang et al., where an oligopeptide from Sepia ink induced the apoptosis of lung cancer cells via the mitochondrial pathway [57]. The antiapoptotic Bcl-2 family proteins prevent cell death induced by various apoptotic stimuli by inhibiting the release of mitochondrial cytochrome C, and inhibiting the activation of caspase-9 and caspase-3 [58]. Thus, a decrease in the expression of Bcl-2 protein coupled with an increase in the expression of Bax often facilitates apoptosis [59,60]. The studies presented thus far provide evidence that suggest peptide B11 exerts an antiproliferative effect in cancer cells by targeting the mitochondria, causing mitochondrial dysfunction through a loss in membrane potential, and consequently inducing mitochondrial-dependent apoptosis.
4. Materials and Methods
4.1. Peptide Synthesis
Peptide B11 (RIRDAIAHGYIVDKV) and rhodamine-labeled B11 were synthesized by a commercial company (Scilight Biotechnology, Beijing, China) using the solid-phase procedure as reported by Huertas et al. [61]. Briefly, 9-fluorenylmethoxycarbonyl amino acid (Fmoc amino acid) and 2,6-dichlorobenzenoylchloride (DCB) were added to the resin to attach the first amino acid. The deprotection was conducted by adding piperidine to remove the protection group, and then the activated amino acid was attached to the peptidyl resin with agitation to couple the next residue. The cycle of deprotection and coupling was repeated until the target peptide was completely synthesized. Rhodamine-labeling was carried out after the last deprotection and coupling step. The peptide was then cleaved from the resin with trifluoroacetic acid (TFA). The disulfide bridge in the peptide was formed by DMF, and C-terminal amidation was conducted by the amidating enzyme. The synthetic peptides were purified by reversed-phase high performance liquid chromatography (RP-HPLC, Omaha, NE, USA) with an Agela C18 column (Agilent, CA, USA). The purity and molecular masses of the purified synthetic peptides were determined using RP-HPLC and MALDI-TOF mass spectrometry (Bruker Daltonics, Bremen, Germany).
4.2. Cell Culture
Human cervical cancer cells (HeLa cells), human hepatocellular carcinoma cells (HepG2 cells), human esophageal cancer cell (EC109 cells), and normal liver cell lines (THLE-3 cells) were kind gifts from Dr. En Min Li of Shantou University Medical College, Shantou University, China. All of the cell lines were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) and 1% penicillin/streptomycin (Gibco), and then maintained in a 5% CO2 incubator at 37 °C.
4.3. Cell Proliferation Assay
Cell proliferation was determined using the MTS assay as reported by Kong et al. [62]. Briefly, 5 × 103 cells were seeded onto 96-well plates overnight. Next, cells were treated with 50 μg/mL of peptide B11 or 50 μg/mL 5-fluorouracil (5-FU) as a positive control, with PBS (0.01 M, pH 7.4) used as the negative control. After 24 h, 20 µL/well of CellTiter 96 AQueous One Solution (MTS) solution (Promega, Madison, WI, USA) was added and incubated at 37 °C for 3 h. Optical density (OD) was measured using a microplate reader (BioTek, Winooski, VT, USA) at 490 nm. The viability rate was calculated as, cell viability% = ODB11/ODPBS × 100%. Triplicate samples were analyzed, and data were represented as means ± standard error (SD). Experiments were repeated at least three times and the p-values were determined using Student’s t-test.
4.4. Cytological Effect of Peptide B11 on HeLa Cells
To examine the cytological effects of peptide B11 on cells, HeLa cells were plated onto 96-well plates (5 × 103/well) overnight, followed by treatment with peptide B11 (50 μg/mL), 5-FU (50 μg/mL), or PBS (0.01 M, pH 7.4) for 24 h. Next, media was removed, and cells were washed two times with PBS (0.01 M, pH 7.4). Following this, cells were fixed for 20 min with 2.5% glutaraldehyde, and then washed twice with PBS (0.01 M, pH 7.4). The fixed cells were stained for 8 min with 1 μg/mL of 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) at 37 °C. Cells were washed four times with PBS (0.01 M, pH 7.4), and finally examined under a fluorescence microscope (Olympus, Tokyo, Japan) as previously described [63].
4.5. Annexin V-FITC/PI Apoptosis Detection Assay
The ability of peptide B11 to induce cell death in terms of apoptosis was determined using flow cytometry with Annexin V-FITC apoptosis detection kit (Beijing Beyotime Corp, Beijing, China) following the manufacturers’ instructions. Briefly, HeLa cells were plated onto 24-well plates (1 × 105/well) overnight. Next, cells were treated with peptide B11 (50 μg/mL), PBS (0.01 M, pH 7.4), and 5-FU (100 μg/mL) for eight to 48 h. At each time point, cells were washed with PBS (0.01 M, pH 7.4) and re-suspended in 195 μL of binding buffer (10 mM of Hepes/NaOH, pH 7.4, 140 mM of NaCl, 2.5 mM of CaCl2); then, five μL of Annexin V-FITC and 10 μL of propidium iodide (PI) were added to stain for 15 min at room temperature in the dark. Cells were analyzed on an Accuri C6 flow cytometer (BD bioscience, SanDiego, CA, USA).
4.6. Prediction of the Structural Characteristics and Features of Peptide B11
The physicochemical properties of peptide B11 were analyzed using the antimicrobial peptide database web server (APD3, Omaha, NE, USA), while the helical properties of peptide B11 were determined by Schiffer Edmundson wheel modeling using the DNAstar Lasergene 7.1 programme. The 3D structure of peptide B11 was predicted using the I-TASSER program (Version 5.1, zhanglab, Ann Arbor, MI, USA) (http://zhanglab.ccmb.med.umich. edu/I-TASSER).
4.7. Cell Uptake Studies
The uptake of peptide B11 by cells was performed according to the method described by Wang et al. [48]. Briefly, HeLa cells (1 × 105) were plated onto a 35-mm glass-bottom dish (In Vitro Scientific, Sunnyvale, CA, USA) overnight, followed by treatment with 50 μg/mL of rhodamine-labeled B11 for eight to 24 h, and placed in a CO2 incubator at 37 °C. The media was removed, and the cells were washed three times with PBS (0.01 M, pH 7.4). Following this, cells were stained with one μg/mL of Hoechst 33342 (Beijing Beyotime Corp) for 10 min at 37 °C, before being washed three times with PBS (0.01 M, pH 7.4). Cells were imaged using a confocal microscope (LSM 800, Carl Zeiss, Jena, Germany).
4.8. Analysis of Subcellular Localization
HeLa cells (1 × 105) were plated onto a 35-mm glass-bottom dish (In Vitro Scientific) overnight, and then treated with 50 μg/mL of rhodamine-labeled B11 for eight h, and placed in a CO2 incubator at 37 °C. The media was removed, and the cells were washed three times with PBS (0.01 M, pH 7.4). Following this, cells were incubated for 30 min with 200 nM of MitoTracker Green (Beijing Beyotime Corp), and then for 10 min with one μg/mL Hoechst 33342 at 37 °C to visualize the mitochondria and nuclei, respectively. Next, the reagents were removed, and the monolayer of cells washed three times with ice-cold PBS before being examined by a confocal laser-scanning microscope (LSM 800, Carl Zeiss, Germany).
4.9. JC-1 Dye Staining for Mitochondrial Membrane Potential Analysis
The loss of mitochondrial membrane potential (∆Ψm) was examined by confocal laser-scanning microscopy (LSM 800, Carl Zeiss, Germany) using 5,5′,6,6′-tetrachloro-1,1′, 3,3′-tetraethylbenzimidazole-carbocyanide iodine (JC-1; Beijing Beyotime Corp) staining. Briefly, HeLa cells were seeded at a density of 1 × 105/well onto a 35-mm glass-bottom dish (In Vitro Scientific). After overnight incubation, cells were treated for 24 h with peptide B11 (50 μg/mL) and PBS (0.01 M, pH 7.4); then, they were placed in a carbon dioxide (CO2) incubator at 37 °C. The spent media was removed, and cells washed once with PBS. Next, one mL of complete culture media and one mL of JC-1 working solution were added to each well, and then they were incubated for 20 min at 37 °C protected from light according to the manufacturer’s instructions. After being washed twice with buffer solution, cells were examined using a laser-scanning confocal microscope (LSM 800, Carl Zeiss, Germany) at room temperature. Cells were treated with 10 μM of carbonyl cyanide m-chlorophenylhydrazone (CCCP), which is a protonophore that can cause the dissipation of ∆Ψm, and was used as positive control.
4.10. Western Blot Analysis
Western blot analysis was performed as described previously [64]. Briefly, HeLa cells were plated onto six-well plates (1 × 106/well) overnight, and then treated with peptide B11 (50 μg/mL) and PBS (0.01 M, pH 7.4) for 24 h and collected, washed, and pelleted by centrifugation. Cell pellets were lysed with Radioimmunoprecipitation assay (RIPA) buffer (150 mM of sodium chloride, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 50 mM of Tris–HCl, pH 7.5, and 2 mM of ethylenediaminetetraacetic acid (EDTA)) with protease inhibitor (Roche, Indianapolis, IN, USA). Total protein extracts were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using a 12% SDS-PAGE gel, and then transferred onto a polyvinylidene difluoride (PVDF) membrane with a wet transfer apparatus according to the manufacturer’s instructions (Bio-Rad, Richmond, CA, USA). The membrane was blocked for 1 h with 5% skimmed milk in Tris Buffered Saline with Tween (TBST) (0.1% Tween-20, 20 mM Tris, 0.15 M NaCl, pH 7.4) at room temperature. Next membranes were probed with primary antibodies: anti-caspase-9 antibody (Rabbit monoclonal, Abcam; 1:1000), anti-caspase-3 antibody (Rabbit monoclonal, Abcam; 1:1000), anti-Bax (Rabbit monoclonal, Abcam; 1:1000), anti-β-actin antibody (Rabbit polyclonal, Abbkine; 1:1000) or anti-Bcl-2 antibody (Mouse monoclonal, Abbkine; 1:1000). After being washed three times with TBST, membranes were then incubated with horseradish peroxidase (HRP) linked goat anti-mouse or goat anti-rabbit secondary antibodies (Sigma-Aldrich, St Louis, MO, USA 1:5000) for 1 h at room temperature. Signals were detected by chemiluminescence using an enhanced chemiluminescence (ECL)-detecting reagent, and images were captured using the GE Amersham Imager 600 imaging system (GE, Boston, MA, USA).
5. Conclusions
Collectively, the data generated in the current study indicates that antimicrobial peptide B11, which is derived from the large molecular weight hemocyanin protein of L. vannamei, has antiproliferative effects on cancer cells, as it was able to cause mitochondrial dysfunction and induce apoptosis. In view of these interesting findings, future studies would explore the antitumor potential of peptide B11 using an animal model.
6. Patents
Zhang YL, Liu SJ, Zheng LY and Wang F have filed a patent that relates to the use of the peptide (B11) for tumor suppression.
Author Contributions
Y.Z. conceived and coordinated the study. S.L., J.J.A. and L.Z. designed, performed and analyzed the experimental data. Z.Z., M.Z. and J.L. contributed reagents/materials and provided technical assistance. Y.Z., S.L., J.J.A. and F.W. wrote the paper. All authors reviewed the results and approved the final version of the manuscript.
Funding
This research was funded by the National Natural Science Foundation of China (Grant numbers [31372558 & 31672689]), Natural Science Foundation of Guangdong Province (Grant number [2017A030311032]), and Department of Education of Guangdong Province (Grant number [2017KZDXM033]).
Conflicts of Interest
The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.
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Sample Availability: Samples of the compounds are not available from the authors as they are patented.
Figure 1. Reverse phase high performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis of L. vannamei hemocyanin-derived antimicrobial peptide B11. (A) Chromatographic profile of purified peptide B11 products, (B) MALDI-TOF-MS spectra of purified peptide B11.
Figure 1. Reverse phase high performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis of L. vannamei hemocyanin-derived antimicrobial peptide B11. (A) Chromatographic profile of purified peptide B11 products, (B) MALDI-TOF-MS spectra of purified peptide B11.
Molecules 23 03202 g001
Figure 2. Inhibitory effects of peptide B11 on growth of immortalized cancer and non-cancer cells. The cancer cell lines (A) human cervical cancer (HeLa) cells, (B) human hepatocellular carcinoma (HepG2) cells, (C) human esophageal cancer (EC109) cells, and immortalized normal human liver cells (D) THLE-3 cells (T-antigen-immortalized human liver epithelial (THLE) cells) were grown for 24 h in the presence of 50 μg/mL of peptide B11. Cell proliferation was analyzed using the MTS assay with PBS used as the negative control, while 5-fluorouracil (5-FU) was used as the positive control. Data represent means ± SD for three independent experiments (n = 3). Significant difference relative to control (PBS-treated cells) were determined by one-way ANOVA and indicated by asterisks (* p < 0.05, ** p < 0.01).
Figure 2. Inhibitory effects of peptide B11 on growth of immortalized cancer and non-cancer cells. The cancer cell lines (A) human cervical cancer (HeLa) cells, (B) human hepatocellular carcinoma (HepG2) cells, (C) human esophageal cancer (EC109) cells, and immortalized normal human liver cells (D) THLE-3 cells (T-antigen-immortalized human liver epithelial (THLE) cells) were grown for 24 h in the presence of 50 μg/mL of peptide B11. Cell proliferation was analyzed using the MTS assay with PBS used as the negative control, while 5-fluorouracil (5-FU) was used as the positive control. Data represent means ± SD for three independent experiments (n = 3). Significant difference relative to control (PBS-treated cells) were determined by one-way ANOVA and indicated by asterisks (* p < 0.05, ** p < 0.01).
Molecules 23 03202 g002
Figure 3. Peptide B11 affects the cell morphology and induces the apoptosis of HeLa cells. (A) Changes in cell morphology following treatment with PBS, peptide B11, and 5-FU for 24 h. Micrographs were obtained using an inverted microscope (20×). (B) Changes in cell nuclei morphology following treatment with PBS, peptide B11, and 5-FC for 24 h. The 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) stained nuclei were observed with a fluorescence microscope (20×). (C) Flow cytometric analysis of apoptosis in HeLa cells after 8 h to 48 h of treatment with PBS, peptide B11, and 5-FU, and staining with Annexin V/propidium iodide (Annexin V/PI). Quadrants: lower-left represent live cells (Annexin V negative/PI negative); lower-right represent early apoptotic/primary apoptotic cells (Annexin V positive/PI negative); upper-right represent late apoptotic/secondary apoptotic cells (Annexin V positive/PI positive); upper-left represent necrotic cells (Annexin V negative/PI positive). The numbers in the respective quadrants indicate the percentage of cells present in that area. Data shown represent one of three independent experiments.
Figure 3. Peptide B11 affects the cell morphology and induces the apoptosis of HeLa cells. (A) Changes in cell morphology following treatment with PBS, peptide B11, and 5-FU for 24 h. Micrographs were obtained using an inverted microscope (20×). (B) Changes in cell nuclei morphology following treatment with PBS, peptide B11, and 5-FC for 24 h. The 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) stained nuclei were observed with a fluorescence microscope (20×). (C) Flow cytometric analysis of apoptosis in HeLa cells after 8 h to 48 h of treatment with PBS, peptide B11, and 5-FU, and staining with Annexin V/propidium iodide (Annexin V/PI). Quadrants: lower-left represent live cells (Annexin V negative/PI negative); lower-right represent early apoptotic/primary apoptotic cells (Annexin V positive/PI negative); upper-right represent late apoptotic/secondary apoptotic cells (Annexin V positive/PI positive); upper-left represent necrotic cells (Annexin V negative/PI positive). The numbers in the respective quadrants indicate the percentage of cells present in that area. Data shown represent one of three independent experiments.
Molecules 23 03202 g003
Figure 4. Predicted structure of peptide B11 and cell uptake analysis. (A) Amino acid sequence and primary structure of peptide B11. (B) The Schiffer–Edmundson helical wheel representation of peptide B11, showing the helical wheel projects and arrangement of amino acid residues counting from the amino (N) terminus. (C) Predicted 3D modeled structure of peptide B11. (D) Uptake of rhodamine-labeled B11 by HeLa cells. Cells were treated with rhodamine-labeled B11 at the indicated time points (0 h, 8 h, and 24 h), stained with Hoechst 33342, and then observed under a confocal microscope (scale bar = 20 µm).
Figure 4. Predicted structure of peptide B11 and cell uptake analysis. (A) Amino acid sequence and primary structure of peptide B11. (B) The Schiffer–Edmundson helical wheel representation of peptide B11, showing the helical wheel projects and arrangement of amino acid residues counting from the amino (N) terminus. (C) Predicted 3D modeled structure of peptide B11. (D) Uptake of rhodamine-labeled B11 by HeLa cells. Cells were treated with rhodamine-labeled B11 at the indicated time points (0 h, 8 h, and 24 h), stained with Hoechst 33342, and then observed under a confocal microscope (scale bar = 20 µm).
Molecules 23 03202 g004aMolecules 23 03202 g004b
Figure 5. Localization of peptide B11 in the mitochondria, its effect on mitochondrial membrane potential (∆Ψm), and apoptosis induction in HeLa cells. (A) Microscopic images showing the intracellular localization of rhodamine-labeled B11 in HeLa cells. Cells were treated with 50 μg/mL of rhodamine-labeled B11 for 8 h, washed with PBS and stained with 200 nM of MitoTracker Green. Images were captured with a confocal microscope under a 40× objective (scale bar = 10 µm). (B) Mitochondrial membrane potential (∆Ψm) of HeLa cells treated with peptide B11. Cells were treated for 24 h with peptide B11 (50 μg/mL) and PBS (0.01 M, pH 7.4), followed by staining with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazole-carbocyanide iodine (JC-1) working solution and incubated for 20 min at 37 °C protected from light. Images were observed under confocal microscopy (scale bar = 20 µm). For positive control, cells were treated with 10 µM of carbonyl cyanide m-chlorophenylhydrazone (CCCP). Red fluorescence represents the mitochondrial aggregate form of JC-1, indicating an intact mitochondrial membrane potential. Green fluorescence represents the monomeric form of JC-1, indicating dissipation of the ∆Ψm. (C) Immunoblots of (i) caspase-9 and caspase-3, (ii) Bax, and (iii) Bcl-2 protein levels in peptide B11-treated HeLa cells analyzed by Western blot. Cell lysates from HeLa cells treated with peptide B11 (50 μg/mL) or PBS (0.01 M, pH 7.4) for 24 h were analyzed using the appropriate antibodies, with β-actin used as a loading control. Numbers below the blots represent the relative gray values determined using ImageJ program.
Figure 5. Localization of peptide B11 in the mitochondria, its effect on mitochondrial membrane potential (∆Ψm), and apoptosis induction in HeLa cells. (A) Microscopic images showing the intracellular localization of rhodamine-labeled B11 in HeLa cells. Cells were treated with 50 μg/mL of rhodamine-labeled B11 for 8 h, washed with PBS and stained with 200 nM of MitoTracker Green. Images were captured with a confocal microscope under a 40× objective (scale bar = 10 µm). (B) Mitochondrial membrane potential (∆Ψm) of HeLa cells treated with peptide B11. Cells were treated for 24 h with peptide B11 (50 μg/mL) and PBS (0.01 M, pH 7.4), followed by staining with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazole-carbocyanide iodine (JC-1) working solution and incubated for 20 min at 37 °C protected from light. Images were observed under confocal microscopy (scale bar = 20 µm). For positive control, cells were treated with 10 µM of carbonyl cyanide m-chlorophenylhydrazone (CCCP). Red fluorescence represents the mitochondrial aggregate form of JC-1, indicating an intact mitochondrial membrane potential. Green fluorescence represents the monomeric form of JC-1, indicating dissipation of the ∆Ψm. (C) Immunoblots of (i) caspase-9 and caspase-3, (ii) Bax, and (iii) Bcl-2 protein levels in peptide B11-treated HeLa cells analyzed by Western blot. Cell lysates from HeLa cells treated with peptide B11 (50 μg/mL) or PBS (0.01 M, pH 7.4) for 24 h were analyzed using the appropriate antibodies, with β-actin used as a loading control. Numbers below the blots represent the relative gray values determined using ImageJ program.
Molecules 23 03202 g005
Table 1. Predicted physicochemical properties of peptide B11.
Table 1. Predicted physicochemical properties of peptide B11.
PropertyParameter
SequenceRIRDAIAHGYIVDKV
Molecular weight (Da)1767
Total hydrophobic ratio46%
Charge at pH 7.0+1
Hydrophobicity <H>0.333
Hydrophobic moment <µH>0.119
Average hydropathy value0.0466
Molar extinction coefficient1490
Isoelectric point8.862
Share and Cite
MDPI and ACS Style
Liu, S.; Aweya, J.J.; Zheng, L.; Wang, F.; Zheng, Z.; Zhong, M.; Lun, J.; Zhang, Y. A Litopenaeus vannamei Hemocyanin-Derived Antimicrobial Peptide (Peptide B11) Attenuates Cancer Cells’ Proliferation. Molecules 2018, 23, 3202. https://doi.org/10.3390/molecules23123202
AMA Style
Liu S, Aweya JJ, Zheng L, Wang F, Zheng Z, Zhong M, Lun J, Zhang Y. A Litopenaeus vannamei Hemocyanin-Derived Antimicrobial Peptide (Peptide B11) Attenuates Cancer Cells’ Proliferation. Molecules. 2018; 23(12):3202. https://doi.org/10.3390/molecules23123202
Chicago/Turabian Style
Liu, Shangjie, Jude Juventus Aweya, Liyuan Zheng, Fan Wang, Zhou Zheng, Mingqi Zhong, Jingsheng Lun, and Yueling Zhang. 2018. "A Litopenaeus vannamei Hemocyanin-Derived Antimicrobial Peptide (Peptide B11) Attenuates Cancer Cells’ Proliferation" Molecules 23, no. 12: 3202. https://doi.org/10.3390/molecules23123202
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2,012,607,942,674,227,500 | Intelligent Design: Coming To A State...
Intelligent Design: Coming To A State Legislature Near You
There are 52857 comments on the www.scientificblogging.com story from May 7, 2008, titled Intelligent Design: Coming To A State Legislature Near You. In it, www.scientificblogging.com reports that:
Religiously motivated strategies to oppose evolution in schools started out with efforts to ban the teaching of evolution. When that approach was struck down by the Supreme Court, Creationists took to arguing that 'Creation Science' was as scientifically legitimate as evolutionary biology, and therefore the two subjects should get equal space in the school curriculum.
Join the discussion below, or Read more at www.scientificblogging.com.
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“Got Science?”
Since: Apr 07
Location hidden
#1 May 7, 2008
I confess the confusion is lost on me and the controversy is pointless but teaching all alternatives because biology cannot explain everything today is just plain bad science.
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Jacques
Pretoria, South Africa
#2 May 8, 2008
Cash wrote:
I confess the confusion is lost on me and the controversy is pointless but teaching all alternatives because biology cannot explain everything today is just plain bad science.
I hear you. This entire debate is a joke. If it happens (ID in schools) well, kiss America goodbye and say hello to the Chinese, taking charge of modern engineering. I wonder how many doctors, engineers, scientists will be brought forth by this system?
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Bohemian
New Zealand
#3 May 8, 2008
Hardly surprising that this is again an issue in America. It merely vindicates the opinion the rest of the word has of many Americans.
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Steve
United States
#4 May 8, 2008
Cash - Got Science?
"I confess the confusion is lost on me
and the controversy is pointless but
teaching all alternatives because
biology cannot explain everything today
is just plain bad science. "
BS - and more of same, and
Piled Higher and Deeper!!!
Wasting time teaching garbage is
bad for kids and bad for society.
Leria
Arlington, VA
#5 May 8, 2008
When are people going to realize that religions (all of them) are lies....... probably never, until we start telling people that they cannot indoctrinate their children with the lies of religion.
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Steve
United States
#6 May 8, 2008
Steve wrote:
Cash - Got Science?
"I confess the confusion is lost on me
and the controversy is pointless but
teaching all alternatives because
biology cannot explain everything today
is just plain bad science. "
BS - and more of same, and
Piled Higher and Deeper!!!
Wasting time teaching garbage is
bad for kids and bad for society.
I agree, the teaching of garbage like Creationism/ID is bad for kids and bad for society. No one can pile it higher or deeper.
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Zeke
Montague, Canada
#7 May 8, 2008
Leria wrote:
... until we start telling people that they cannot indoctrinate their children with the lies of religion.
In the meantime we need to make sure they don't indoctrinate *other people's* children in the public schools.
ed crandall
United States
#9 May 8, 2008
I can tell "inteligent design" has been wasted on most of the comments that I've read here. A thinking peson Knows that nothing just happens, all things are the result of a former action.(Intelligence)
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Bud
Tallahassee, FL
#10 May 8, 2008
ed crandall wrote:
I can tell "inteligent design" has been wasted on most of the comments that I've read here. A thinking peson Knows that nothing just happens, all things are the result of a former action.(Intelligence)
So let's see the evidence that supports your hypothesis, Crandall. Or how about an experiment to test it? No?
ID is not science.
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Steve
United States
#11 May 8, 2008
Bud wrote:
<quoted text>
So let's see the evidence that supports your hypothesis, Crandall. Or how about an experiment to test it? No?
ID is not science.
I'm all for that. I'd like to see the evidence that scientifially supports the so-called inteelligent design.
Cookie
Maryland Heights, MO
#12 May 8, 2008
I do not understand how anyone can think the earth is 6000 years old and man appeared out of the blue. Whether or not there is a higher being no one can ever know, you don't find out until you are dead, and then you can't tell anyone. ID is just junk. And the US is on the path to mediocre schools and dumber people.
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formally abducted
United States
#13 May 8, 2008
did'nt the taliban start out like this,and so,the jihad begins.
“"GUILTY"”
Since: Apr 08
United States
#14 May 8, 2008
That which the Government subsidizes, it can control.
Now we know why there is so much federal funding being thrown at schools... It is not supposed to be this way folks!
CURTYMACK
United States
#15 May 8, 2008
Creationism is a religious ideology. Lord only knows how many different religions and versions of creation exist. Consequently, creationism should be taught by religious instructors not public school teachers. No religion proves the existence of God. Likewise evolution theory does not disprove the existence of a God.
Teachers should emphasize that evolution is merely a theory that attempts to explain how life could have began and could have changed over time.
Plenty of scientist understand evolution theory yet still believe in a God. So if a parent fears their child learning of this theory, it speaks volumes about their religious instructors.
Tacky
United States
#16 May 8, 2008
"So let's see the evidence that supports your hypothesis, Crandall. Or how about an experiment to test it? No?"
This is the exact problem with either intelligent design or with evolution. The evidence is very questionable and there is no experiment to put the theory to the test. If evolution is as well documented as some believe, why are these supporters so opposed to any criticism of the theory? It is indeed a theory, not established fact.
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“make mine a double, please”
Since: Sep 07
chicago
#17 May 8, 2008
KTM wrote:
Just more idiocy brought to America by the republican moron camo-crowd.
that's just stupid.
Kirlak
New York, NY
#18 May 8, 2008
US Constitution still guarantees separation of Church and State. I believe in God, but I also strongly believe that all regular teaching in Schools must be separate from Sunday School teachings. Bringing religion into a regular School is ABSOLUTELY AGAINST EVERY FREEDOM established by the founding fathers back over 200 years ago. Any politician, even hinting about breaking this separation, must be kicked out of politics immediately.
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an american
United States
#19 May 8, 2008
What about this?... Life evolved from the big bang. That life became intelligent, because its nature was spiritual. That life observed evolution was happening all around and decided to find his own isolated place to be. That life found a little blue planet and decided to stay and observe the evolution.
an american
United States
#20 May 8, 2008
This way, everyone gets to be right.
R Bratton
United States
#21 May 8, 2008
It takes more faith to believe in evolution than in creation! Someone has asked for evidence of evolution. Well, it is obvious, man is evolving backwards and apart fromt the Bible, you cannot explain what is going on in the world today. God does not always payoff on Friday night, but He pays off! Payday is not far off!
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"label": "Not Applicable/Indeterminate"
},
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}
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"label": "General Audience"
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"code": "2",
"label": "High School Level"
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} | 57 | ff2a0dd8e83e1035faba2b053f3eae93 |
-6,797,666,868,164,174,000 | guide
Contributing GO Annotations
If your research group has GO annotations for a species that is not currently included in the GO, whether or not these annotations cover the entire genome, or if your research team has identified gaps or inaccuracies in the current set of GO annotations, this guide is for you. Choose the scenario that best describes your research group and follow the steps as indicated in the following pages.
Contributing to GO
Research groups may contribute to the Gene Ontology Consortium (GOC) by providing suggestions for updating the ontology (e.g. requests for new terms) or by providing annotations, that is, associations between genes or gene products and ontology terms. Suggested edits are reviewed by the ontology editors and implemented where appropriate.
The following pages explain how you can contribute to the project. Please begin by choosing whether you wish to contribute annotations or terms to the Gene Ontology.
GO Annotation File (GAF) Format 2.1
Annotation data is submitted to the GO Consortium in the form of gene association files, or GAFs. This guide lays out the format specifications for GAF 2.1; for the previous GAF 2.0 file syntax, please see the GAF 2.0 file format guide.
For the first GAF 1.0 file syntax, please see the GAF 1.0 file format guide.
Please see the information on the changes in GAF 2.1.
GO Mailing Lists
Contact GO
The Gene Ontology project very much encourages input from the community into both the content of the GO and annotation using GO. We are very happy to work with others to ensure that the GO is both complete and accurate, and we also very much encourage communities to submit GO annotations for inclusion in the GO database. Please contact us.
GO Consortium Photo Album
• GO Consortium Meeting, Hôtel Mont Gabriel, Sainte-Adèle, Québec, Canada, October 21–23, 2008
• GO Consortium Meeting, Jesus College, Cambridge University, Cambridge, UK, January 8–10, 2007
• Annotation Camp, Stanford University, Palo Alto, California, USA, July 12–14, 2006
• Annotation Workshop, Stanford University, Palo Alto, California, USA, July 10–11, 2006
• GO Consortium Meeting, St. Croix, US Virgin Islands, March 31–April 2, 2006
Authoritative Database Groups
Authoritative Database Groups Where two or more databases are submitting data on the same species the GO Consortium encourages the model whereby one database group collects all annotation data for that species, removes the redundant (duplicate) annotations, and then submits the total dataset to the central repository. This ensures that no redundant annotations will appear in the master dataset. The table below documents those species for which a single database group is responsible for collating and submitting annotations.
Authoritative Database Groups
Authoritative Database Groups Where two or more databases are submitting data on the same species the GO Consortium encourages the model whereby one database group collects all annotation data for that species, removes the redundant (duplicate) annotations, and then submits the total dataset to the central repository. This ensures that no redundant annotations will appear in the master dataset. The table below documents those species for which a single database group is responsible for collating and submitting annotations.
External Mapping File Format
Mappings of GO have been made to other many other classification systems; a full list is available on the Mappings to GO page. This page describes the format of these files.
Format Specification
The source of the external file is given in the line beginning !Uses: !Uses:http://www.tigr.org/docs/tigr-scripts/egad_scripts/role_reports.spl, 15 aug 2000.
The line syntax for mappings is:
external database:term identifier (id/name) > GO:GO term name ; GO:id
For example:
Temporal information in Annotation Extensions
Adding temporal information
Using biological process terms to enhance cellular component annotations
Cellular component annotations can be enhanced by specifying that localization is observed during a specific biological process, e.g. a cell cycle phase. These annotations are constructed by putting the exists_during relationship and the identifier from the biological process ontology in the annotation extension column.
Use cases
1. A gene product is localized to the nuclear periphery in S phase, G2, and mitosis (S. pombe Ulp1; PMID:11884512)
Spatial information in Annotation Extensions
Adding spatial information
Usefulness of capturing cell type or tissue type specific location of action
Investigative methods that work solely with a specific tissue or cell type (such as laser capture microdissection) are becoming more commonplace and allow for downstream genetic or proteomic analyses that are not contaminated by surrounding tissue. In addition the separation of subcelluar particles via cell fractionation techniques enables the study of the constituents of a particular cell part/organelle. | {
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-5,091,271,675,622,259,000 | Buy a SimpleChIP Kit and Try our New SimpleChIP® Universal qPCR Master Mix (88989P) for Free | Get The Code >>
Product Results
Confocal immunofluorescent image #9102
Primary Antibodies from CST
Our primary antibodies have been tested and validated on the lot level by our in-house team of scientists for specificity, sensitivity, and reproducibility. Choose from a list of proteins below to further refine your search results or use the faceting options on the left to select the attributes you want in your antibody.
# Product Name Application Reactivity
H, M, R, All
H, All
H, M, R, All
H, M, Dm, All
All
All
All
H, M, R, Mk
H, M, R
H, B, Pg
H, M, R, Mk
H
H, M
H
H, M, Mk
H, M, R, Mk, All
H, M, Mk, All
H, M, R, Mk, Sc, All
H, M, R, All
All
H
H, All
All
H, All
All
H, M, R, Mk, All
All
All
All
H, M, All
Results per page: | {
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} | 57 | ff2a0dd8e83e1035faba2b053f3eae93 |
-1,451,576,242,228,048,600 | The world's first wiki where authorship really matters (Nature Genetics, 2008). Due credit and reputation for authors. Imagine a global collaborative knowledge base for original thoughts. Search thousands of articles and collaborate with scientists around the globe.
wikigene or wiki gene protein drug chemical gene disease author authorship tracking collaborative publishing evolutionary knowledge reputation system wiki2.0 global collaboration genes proteins drugs chemicals diseases compound
Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
Dual roles of tropomyosin as an F-actin stabilizer and a regulator of muscle contraction in Caenorhabditis elegans body wall muscle.
Tropomyosin is a well-characterized regulator of muscle contraction. It also stabilizes actin filaments in a variety of muscle and non-muscle cells. Although these two functions of tropomyosin could have different impacts on actin cytoskeletal organization, their functional relationship has not been studied in the same experimental system. Here, we investigated how tropomyosin stabilizes actin filaments and how this function is influenced by muscle contraction in Caenorhabditis elegans body wall muscle. We confirmed the antagonistic role of tropomyosin against UNC-60B, a muscle-specific ADF/cofilin isoform, in actin filament organization using multiple UNC-60B mutant alleles. Tropomyosin was also antagonistic to UNC-78 (AIP1) in vivo and protected actin filaments from disassembly by UNC-60B and UNC-78 in vitro, suggesting that tropomyosin protects actin filaments from the ADF/cofilin-AIP1 actin disassembly system in muscle cells. A mutation in the myosin heavy chain caused greater reduction in contractility than tropomyosin depletion. However, the myosin mutation showed much weaker suppression of the phenotypes of ADF/cofilin or AIP1 mutants than tropomyosin depletion. These results suggest that muscle contraction has only minor influence on the tropomyosin's protective role against ADF/cofilin and AIP1, and that the two functions of tropomyosin in actin stability and muscle contraction are independent of each other. Cell Motil. Cytoskeleton 2006. (c) 2006 Wiley-Liss, Inc.[1]
References
WikiGenes - Universities
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3,646,084,271,142,257,700 | Research & development
Laser to break specific molecular bonds
19 May, 2006
A team of researchers has achieved a long-sought scientific goal: using laser light to break specific molecular bonds.
First man-made fractal molecule
12 May, 2006
All objects in nature are made of irregular molecules called fractals and scientists now have created and captured an image of the largest man-made fractal molecule at the nanoscale.
Sugar in cell communication
10 May, 2006
A research team from Uppsala University has uncovered an entirely new mechanism for how communication between cells is regulated.
Sound way to measure fluid in the brain
27 April, 2006
Scientists have developed a way of measuring levels of fluid in the brain that avoids the need for painful lumbar punctures.
Improvement in poultry quality
26 April, 2006
A test has been developed that can assess the robustness of the immune system in poultry by the Eureka E! 2692 molecular tests project.
Delving into the pump mechanism mystery
07 April, 2006
Researchers at the Institute of Biotechnology of the University of Helsinki have identified an internal electron transfer reaction that initiates the proton pump mechanism of the respiratory enzyme.
Stem cells used to make new tendons
06 April, 2006
Weekend athletes who overexert themselves running or playing basketball may one day reap the benefits of research at the Hebrew University of Jerusalem that shows that adult stem cells can be used to make new tendon or ligament tissue.
Critical gene for immune cell activation found
04 April, 2006
Every time the human body encounters a virus, bacteria or other infectious agent, immune cells called B-lymphocytes multiply in lymph nodes and then swing into action to fight off the intruders.
Nanoparticles good for biomedical applications
04 April, 2006
Nanoparticles of gold can act as tiny, precise and powerful heaters, which potentially could be used in biomedical applications, according to a new study.
Aussies helping the fight against bird flu
21 March, 2006
CSIRO scientists have boosted efforts to help stop the spread of bird flu in Asia and thereby reduce the risk of it reaching Australia.
Help for forensics with full-colour fingerprints
16 March, 2006
In the future, forensic investigators may take full-colour fingerprints using new technology developed by a University of Toronto-led team of international researchers.
Coeliac disease research tool
08 March, 2006 | Supplied by: PerkinElmer Pty Ltd
Delfia probes, specific for HLA alleles, provide a research tool for investigating genes involved in predisposition to coeliac disease.
Australians to tap into crop testing in Europe
02 March, 2006
A company has been established in Adelaide to develop scientific techniques for genetic crop testing in Europe.
Study into formation of an embryo
24 February, 2006
A discovery from University of Missouri-Columbia scientists about the formation of an embryo and a placenta before implantation could explain why cloning often fails in farm and laboratory animals.
Developing resistant wheat varieties
23 February, 2006
A new $5 million grant to wheat breeders could shorten the time between the outbreak of diseases and the development of resistant wheat varieties, said the Texas Agricultural Experiment Station state wheat breeder.
• All content Copyright © 2020 Westwick-Farrow Pty Ltd | {
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8,829,063,702,760,420,000 | OriGene Technologies, Inc.
Search:
Left ProductsProducts divider ServicesServices divider technologyTechnology divider researchResearch divider TechsupportTechSupport divider AboutAbout Right
Home Antibody All anti-PTPN7 antibodies
Anti-PTPN7 TRUEMAB Antibody Clone OTI3B10
TrueMAB™ Antibodies - Made against Authentic Protein Antigens
div
Specifications Citations (0) Related Products Product Documents
SKU Description Amount Price Availability*
TA503714 PTPN7 (HePTP) mouse monoclonal antibody, clone OTI3B10 (formerly 3B10) 100ul $379 In Stock
LC409137 PTPN7 HEK293T cell transient overexpression lysate (as WB positive control) 20ug $50 In Stock
CF503714 Carrier-free (BSA/glycerol-free) PTPN7 mouse monoclonal antibody, clone OTI3B10 (formerly 3B10) 100ug $450 3-4 weeks
Add to Shopping Cart
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WB(1)
IF(1)
spacer
Also for PTPN7 (NM_080588)
cDNA Clone shRNA/siRNA CRISPR KO Kit Protein Antibody
OriGene Data
ImmunogenFull length human recombinant protein of human PTPN7(NP_542155) produced in HEK293T cell.
Clone NameClone OTI3B10 IsotypeIgG2b
Species ReactivityHuman , Dog , Rat , Mouse Concentration0.72 mg/ml
Guaranteed Application *WB, IF Suggested DilutionsWB 1:500~2000, IF 1:100,
Predicted MW Explanation 44.8 kDa
BufferPBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide.
Purification Purified from mouse ascites fluids by affinity chromatography
Reference Data
Target NameHomo sapiens protein tyrosine phosphatase, non-receptor type 7 (PTPN7), transcript variant 2
Alternative NameBPTP-4; HEPTP; LC-PTP; LPTP; PTPNI
Database LinkNP_542155
Entrez Gene 5778 Human
Entrez Gene 320139 Mouse
Entrez Gene 246781 Rat
Entrez Gene 607227 Dog
FunctionThe protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. This gene is preferentially expressed in a variety of hematopoietic cells, and is an early response gene in lymphokine stimulated cells. The non-catalytic N-terminus of this PTP can interact with MAP kinases and suppress the MAP kinase activities. This PTP was shown to be involved in the regulation of T cell antigen receptor (TCR) signaling, which was thought to function through dephosphorylating the molecules related to MAP kinase pathway. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq].
Related PathwayPhosphataseDruggable Genome MAPK signaling pathway
* Availability is in business days
* OriGene provides validated application data and protocol, with money back guarantee.
HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY PTPN7 (RC216254, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-PTPN7.
spacer
Anti-PTPN7 mouse monoclonal antibody (TA503714) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY PTPN7(RC216254).
spacer
spacer
Inc 5000 Healthcare Company
All Products by: Title | Price | Category | Popularity | Best Sellers Topselling Products by: Title | Price | Category | Popularity | Favorites
Popular Categories: Popularity | Our Choices | All-Round Favorites | Title Topselling Categories: Popularity | Our Choices | All-Round Favorites | Title | {
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387,041,995,197,835,400 | string(63) "Failed to connect to 205.234.153.91 port 1165: No route to host" How Do Trees Help The Environment Science? - Okela
how do trees help the environment science?
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how do trees help the environment science? community answers | {
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-6,294,094,811,693,112,000 | About Help FAQ
Picalm Gene Detail
Summary
• Symbol
Picalm
• Name
phosphatidylinositol binding clathrin assembly protein
• Synonyms
fit1, fit-1, MGC:19382, MGC:36413, MGC:36616
• Feature Type
protein coding gene
• IDs
MGI:2385902
NCBI Gene: 233489
• Gene Overview
MyGene.info: PICALM
Location & Maps
more
• Sequence Map
Chr7:90130213-90213465 bp, + strand
• From VEGA annotation of GRCm38
Mouse Genome Browser
• Download
Sequence
83253 bp ± kb flank
• Genome Browsers
• Genetic Map
Chromosome 7, 50.47 cM
• Mapping Data
6 experiments
Homology
more
• Human Ortholog
PICALM, phosphatidylinositol binding clathrin assembly protein
• Vertebrate Orthologs
10
• Human Ortholog
PICALM, phosphatidylinositol binding clathrin assembly protein
Orthology source: HomoloGene, HGNC
• Synonyms
CALM, CLTH, LAP
• Links
NCBI Gene ID: 8301
neXtProt AC: NX_Q13492
• Chr Location
11q14.2; chr11:85957171-86069881 (-) GRCh38.p7
Human Diseases
less
• Mutations/Alleles
1 with disease annotations
• References
1 with disease annotations
Mutations,
Alleles, and
Phenotypes
less
• Phenotype Summary
42 phenotypes from 8 alleles in 7 genetic backgrounds
14 phenotypes from multigenic genotypes
1 images
41 phenotype references
Phenotype Overview
adipose tissue
behavior/neurological
cardiovascular system
cellular
craniofacial
digestive/alimentary system
embryo
endocrine/exocrine glands
growth/size/body
hearing/vestibular/ear
hematopoietic system
homeostasis/metabolism
integument
immune system
limbs/digits/tail
liver/biliary system
mortality/aging
muscle
nervous system
pigmentation
renal/urinary system
reproductive system
respiratory system
skeleton
taste/olfaction
neoplasm
vision/eye
Click cells to view annotations.
• All Mutations and Alleles
30
• Chemically induced (ENU)
5
• Chemically induced (other)
1
• Gene trapped
17
• Radiation induced
3
• Targeted
4
• Genomic Mutations
4 involving Picalm
• Incidental Mutations
Mice homozygous for different ENU-induced mutations or knock-out alleles are small, runted and display anemia of variable severity.
Gene Ontology
(GO)
Classifications
less
• All GO Annotations
• GO References
Molecular Function
carbohydrate derivative binding
cytoskeletal protein binding
DNA binding
enzyme regulator
hydrolase
ligase
lipid binding
oxidoreductase
receptor
receptor binding
RNA binding
transcription
transferase
transporter
Biological Process
nucleic acid-templated transcription
carbohydrate derivative metabolism
cell death
cell differentiation
cell proliferation
cellular component organization
establishment of localization
homeostatic process
immune system process
lipid metabolic process
protein metabolic process
response to stimulus
signaling
system development
Cellular Component
cell projection
cytoplasmic vesicle
cytoskeleton
cytosol
endoplasmic reticulum
endosome
extracellular region
Golgi apparatus
mitochondrion
non-membrane-bounded organelle
nucleus
organelle envelope
organelle lumen
plasma membrane
synapse
vacuole
Click cells to view annotations.
Expression
less
Expression Overview
early conceptus
embryo ectoderm
embryo endoderm
embryo mesoderm
embryo mesenchyme
extraembryonic component
alimentary system
auditory system
branchial arches
cardiovascular system
connective tissue
endocrine system
exocrine system
hemolymphoid system
integumental system
limbs
liver and biliary system
musculoskeletal system
nervous system
olfactory system
reproductive system
respiratory system
urinary system
visual system
Click cells to view annotations.
• Assay Results
• Tissues
• cDNA Data
• Literature Summary
Interactions
less
Sequences &
Gene Models
less
Representative SequencesLengthStrain/SpeciesFlank
genomic OTTMUSG00000059751 VEGA Gene Model | MGI Sequence Detail 83253 C57BL/6J ± kb
transcript OTTMUST00000146466 VEGA | MGI Sequence Detail 8209 Not Applicable
polypeptide OTTMUSP00000076257 VEGA | MGI Sequence Detail 660 Not Applicable
For the selected sequence
Polymorphisms
less
• SNPs within 2kb
406 from dbSNP Build 142
Protein
Information
less
Molecular
Reagents
less
• All nucleic 18
Genomic 2
cDNA 13
Primer pair 1
Other 2
Microarray probesets 4
Other
Accession IDs
less
MGD-MRK-9805, MGD-MRK-9806, MGI:95538
References
more
• Summaries
All 55
Developmental Gene Expression 3
Diseases 1
Gene Ontology 10
Phenotypes 41
• Earliest
J:10283 Rinchik EM, et al., A strategy for fine-structure functional analysis of a 6- to 11-centimorgan region of mouse chromosome 7 by high-efficiency mutagenesis. Proc Natl Acad Sci U S A. 1990 Feb;87(3):896-900
• Latest
J:238763 Choo HJ, et al., Karyopherin Alpha 1 Regulates Satellite Cell Proliferation and Survival by Modulating Nuclear Import. Stem Cells. 2016 Jul 19;
Contributing Projects:
Mouse Genome Database (MGD), Gene Expression Database (GXD), Mouse Tumor Biology (MTB), Gene Ontology (GO), MouseCyc
Citing These Resources
Funding Information
Warranty Disclaimer & Copyright Notice
Send questions and comments to User Support.
last database update
02/16/2017
MGI 6.07
The Jackson Laboratory | {
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1,225,318,795,746,644,700 | OriGene Technologies, Inc.
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Anti-CXCR3 Antibody
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TA313938 Rabbit polyclonal anti-CXCR3 antibody 100ul $325 3 Weeks
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OriGene Data
ImmunogenThe antiserum was produced against synthesized peptide derived from internal of human CXCR3.
Clone Name IsotypeIgG
Species ReactivityHuman Concentration1mg/ml
Guaranteed Application *WB Suggested DilutionsWB: 1:500~1:3000, ELISA: 1:5000
BufferPhosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Purification The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Note CXCR3 antibody detects endogenous levels of total CXCR3 protein.
Reference Data
Target NameHomo sapiens chemokine (C-X-C motif) receptor 3 (CXCR3), transcript variant 2
Alternative NameCD182; CD183; CKR-L2; CMKAR3; GPR9; IP10-R; Mig-R; MigR
Database LinkNP_001136269
Entrez Gene 2833 Human
Function
Related PathwayGPCRTransmembraneDruggable Genome Cytokine-cytokine receptor interactionChemokine signaling pathway
* Availability is in business days
* OriGene provides validated application data and protocol, with money back guarantee.
WB Image
Western blot analysis of extracts from K562 cells, using CXCR3 antibody.The lane on the right is treated with the synthesized peptide.
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5,007,596,549,633,786,000 | Landscape Online
About
Landscape Online is a self-published peer reviewed open access journal, associated to the International Association for Landscape Ecology (IALE). Publishing Editor is the German chapter IALE-D. Landscape Online focuses on studies dealing with research in landscape ecology. The subject matter deals with any scientific, educational or applied aspect of processes, dynamics, indicators, controllers and visions related to landscapes. Furthermore, Landscape Online emphasizes the coupling of societal and natural systems, not only the involvement of human impact on landscape systems but also human perception of the landscape, its values and the evaluation of landscapes. Moreover, articles are appropriate that deal with landscape theory, system approaches and conceptual models of landscape, both their improvement and their discussion. Papers may be undisciplinary or multidisciplinary but have interdisciplinary or transdisciplinary appeal.
Landscape Online offers to publish ‘research articles’, ‘short papers’, and ‘reviews’. Furthermore, it offers the article category ‘ideas’ to present new concepts, methods, methodologies, models, approaches, or theoretical aspects in landscape ecology that should be further discussed within the scientific community, and ‘syntheses’ for more specifically target scientific outputs for knowledge brokerage, putting emphasis on theory-practice transfer and policy making.
Please visit the journal website for more information: www.Landscape-Online.org
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-8,473,545,175,785,785,000 | Local BBC Sites
Page last updated at 14:49 GMT, Monday, 21 June 2010 15:49 UK
Night in the Museum Part 4
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Americas Africa Europe Middle East South Asia Asia Pacific | {
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-7,838,101,092,409,852,000 | The Overlord's Muse of
Mother Knows Best State
Overview Factbook Policies People Government Economy Rank Trend Nuke Control Cards
Environmental Beauty
The Master Primotav 1,883.84
Pounds Of Wildlife Per Square Mile
World Census researchers spent many arduous weeks lying on beaches and trekking through rainforests to compile a definitive list of the most attractive and best cared for environments.
Add: Other... | {
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