Glycine dehydrogenase (decarboxylating)

Glycine decarboxylase also known as glycine cleavage system P protein or glycine dehydrogenase is an enzyme that in humans is encoded by the "GLDC" gene.
Reaction.
Glycine decarboxylase () is an enzyme that catalyzes the following chemical reaction:
Thus, the two substrates of this enzyme are glycine and H-protein-lipoyllysine, whereas its two products are H-protein-S-aminomethyldihydrolipoyllysine and CO2.
This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH-NH2 group of donors with a disulfide as acceptor. This enzyme participates in glycine, serine and threonine metabolism. It employs one cofactor, pyridoxal phosphate.
Function.
Glycine decarboxylase is the P-protein of the glycine cleavage system in eukaryotes. The glycine cleavage system catalyzes the degradation of glycine. The P protein binds the alpha-amino group of glycine through its pyridoxal phosphate cofactor. Carbon dioxide is released and the remaining methylamine moiety is then transferred to the lipoamide cofactor of the H protein.
Degradation of glycine is brought about by the glycine cleavage system, which is composed of four mitochondrial protein components: P protein (a pyridoxal phosphate-dependent glycine decarboxylase), H protein (a lipoic acid-containing protein), T protein (a tetrahydrofolate-requiring enzyme), and L protein (a lipoamide dehydrogenase).
Clinical significance.
Glycine encephalopathy is due to defects in "GLDC" or "AMT" of the glycine cleavage system.


Psynapse


Danimer Scientific

Danimer Scientific, formerly known as Meredian Holdings Group Inc. and MHG, is a biopolymer manufacturer headquartered in Bainbridge, Georgia.
Danimer Scientific owns the patent for Nodax medium-chain-length branched polyhydroxyalkanoates, mcl-PHA. The company uses PHA and other biopolymers to create a range of applications such as additives, aqueous coatings, extrusion coating, extrusion lamination, fibers, film resins, hot melt adhesives, injection molding, thermoforming and wax replacement polymers. In addition, Danimer Scientific offers research and development in the formulation of biopolymers. Danimer Scientific also provides toll manufacturing and compounding services, allowing partners to use the Bainbridge facility to manufacture products.
Background.
DaniMer Scientific.
DaniMer Scientific was formed in 2004 to create biodegradable and sustainable solutions to the global dependency on traditional plastics, by using biopolymer materials such as polylactic acid (PLA). DaniMer Scientific specializes in customizing biopolymer formulations.
The company received the Sustainability Award from the Specialty Coffee Association of America (SCAA) in 2007 for the Ecotainer coffee cup, which used a plastic biopolymer derived from corn for the inner lining.
In 2006, the Small Business Innovation Research (SBIR) program awarded Danimer with a Phase 1 monetary grant of $80,000 for the "Creation of Cost Competitive Biodegradable Films from Renewable Resources for Agriculture". The following year the SBIR awarded the company with a Phase 2 monetary grant of $346,000 for the same title, stating that the project's objective was to use renewable materials to develop agricultural mulch films that could adequately degrade before the next growing season
In 2010, DaniMer Scientific received a contract for $846,828 from the organization, Research Partnership to Secure Energy for America (RPSEA) to develop an environmentally friendly fracturing treatment for hydrocarbon production, now known as SqueezeFrac.
September 2011 Danimer Scientific and minority business enterprise, Sijo Global Partners, formed a strategic affiliation where Sijo would offer Danimer’s bioplastic products to market leaders and brand owners.
In January 2013 DaniMer and Henkel formed a partnership to further develop bio-based hot melt adhesives for polyethylene terephthalate (PET) container packaging that are made with 50% renewable content or higher.
Meredian Inc..
Originally formed in 2004, Meredian Inc. manufactures PHA for Danimer Scientific. Danimer Scientific's Nodax PHA is a specialized, medium-chain-length branched polyhydroxyalkanoate, mcl-PHA.
Danimer and Meredian originally purchased Procter & Gamble’s intellectual property on PHA technology in 2007. In 2013, Dr. Isao Noda joined Danimer / Meredian Inc. after spending thirty years with Procter & Gamble.
In 2014 the company received a Food and Substance Contact Notification approval from the United States Food and Drug Association (FDA). The sole-verification guarantees that the biopolymers are safe to use for food contact and are classified as non-hazardous waste after disposal.
Previous Merger into Meredian Holdings Group, Inc..
In February 2014, DaniMer Scientific and Meridian Inc. merged into MHG.
Championed by Dr. Paul Pereira who was CEO and Executive Chairman during the period from September 2013 until October 2015
Corporate Identity Change to Danimer Scientific.
The company's name changed to Danimer Scientific was officially announced in October 2016.
Danimer Scientific PHA.
Danimer Scientific's PHA has received several certifications for biodegradability including: anaerobic and aerobic digestion in soil, freshwater, marine, industrial and home composting.
The patented version of Danimer Scientific's mcl-PHA is known as Nodax, and it is the primary product developed for commercial manufacturing. Danimer Scientific's Nodax PHA is a class of bioplastics produced from bacterial microorganisms that store PHA polyesters for energy in their cell walls. These microbes feed upon plant-based oils procured from non-food sources.
Microorganism & Fermentation.
Plant based oils are transferred to Danimer Scientific’s proprietary bio-reactors. There, the oil is fed to naturally regenerating, soil borne bacteria who produce PHA through biosynthesis.
Reactive Extrusion.
The reactive extrusion process mixes and compounds the bioplastic materials in a unique proprietary procedure that creates a customized biopolymer resin pellet.


Chlorophyll b

Chlorophyll "b" is a form of chlorophyll. Chlorophyll "b" helps in photosynthesis by absorbing light energy. It is more soluble than chlorophyll "a" in polar solvents because of its carbonyl group. Its color is green, and it primarily absorbs blue light.
In land plants, the light-harvesting antennae around photosystem II contain the majority of chlorophyll "b". Hence, in shade-adapted chloroplasts, which have an increased ratio of photosystem II to photosystem I, there is a higher ratio of chlorophyll "b" to chlorophyll "a". This is adaptive, as increasing chlorophyll "b" increases the range of wavelengths absorbed by the shade chloroplasts.
Biosynthesis.
The Chlorophyll "b" biosynthetic pathway utilizes a variety of enzymes. In most plants, chlorophyll is derived from glutamate and is synthesised along a branched pathway that is shared with heme and siroheme.
The initial steps incorporate glutamic acid into 5-aminolevulinic acid (ALA); two molecules of ALA are then reduced to porphobilinogen (PBG), and four molecules of PBG are coupled, forming protoporphyrin IX.
Chlorophyll synthase is the enzyme that completes the biosynthesis of chlorophyll "b" by catalysing the reaction
This forms an ester of the carboxylic acid group in chlorophyllide "b" with the 20-carbon diterpene alcohol phytol.


M. Afzal Hossain

M. Afzal Hossain (born 30 November 1952) is a Bangladeshi academic and scientist in the field of agriculture and biochemistry. He has been serving as vice-chancellor of Hajee Mohammad Danesh Science & Technology University, Dinajpur, since September 2008. In 1998, he was awarded an invitation fellowship from Japan Society for the Promotion of Science (JSPS) to serve as a visiting professor at Hokkaido University, Japan.
Biography.
Early life.
Hossain was born on 30 November 1952 in Majbari village, Shariakandi of Bogra District to Sabder Rahman and Jamila Khatoon. He obtained B.Sc. A.H. (Hons.) majoring in dairy science and M.Sc. A.H. in biochemistry from Bangladesh Agricultural University securing first class in both the cases.
Academic career.
Hossain joined as research associate in 1977 and after as lecturer of biochemistry at BAU in 1978. He was promoted to the rank of assistant professor in 1980. He obtained his PhD degree from National Dairy Research Institute, Haryana, India in 1984 and was promoted to the rank of associate professor in 1986. He was selected for a USAID fellowship and performed postdoctoral research from the Department of Food Science and Nutrition, the University of Hawaii in the United States in 1987. He was promoted to the rank of professor in 1992.
Hossain published his fundamental research papers in several peer-reviewed international and national journals and hence selected for the FAOB fellowship to participate 7th FAOB conference in Malaysia in 1987. Furthermore, he was selected for the European Economic Community postdoctoral fellowship and worked for one year in the Institute of Cellular and Molecular Pathology, Brussels, Belgium in 1992.
Hossain also served as the head of the department of biochemistry for two tenures. He had an ample opportunity to interact with eminent scientists at national and international levels. He participated and presented his research findings at many forums. In this connection, he visited several leading educational and research institutes in different countries of the world. Based on his academic and research potentiality at molecular level he was honoured to avail Jawaharlal Nehru Centre for Advanced Scientific Research-Committee on Science and Technology in Developing Countries Fellowship in 1997 to work as a visiting scientist for three months in Central Food Technological Research Institute, India.
In 1998, Hossain was awarded an Invitation Fellowship from Japan Society for the Promotion of Science (JSPS) to serve as a Visiting Professor at Hokkaido University, Japan. His excellent training in and background of molecular level works combined with his aptitude for higher research imparted him necessary capability to handle several research projects efficiently. He has good number of students received their masters and Ph D degree under his supervision. He has been involved in many professional activities at home and abroad.


Polaribacter

Polaribacter is a Gram-negative, heterotrophic, psychrophilic or mesophilic and aerobic bacteria genus from the family of Flavobacteriaceae.


MADAM-6

MADAM-6, or 2,"N"-dimethyl-4,5-methylenedioxyamphetamine, is a lesser-known recreational drug of the methamphetamine class, similar in structure to MDMA (ecstasy). MADAM-6 was first synthesized by Alexander Shulgin. In Shulgin's book "PiHKAL", the minimum dosage is listed as greater than 280 mg, and the duration is unknown. MADAM-6 produces few to no effects and Shulgin describes it as "not active". Very little data exists about the pharmacological properties, metabolism, and toxicity of MADAM-6.
MADAM-6 has been studied for its potential antiparkinsonian effects. However, there are no clinical trials suggesting the drug is effective against Parkinson's disease.


Linda Griffith

Linda Gay Griffith (born August 30, 1960 Atlanta, Georgia) is an American biological engineer, and Professor of Biological Engineering and Mechanical Engineering at Massachusetts Institute of Technology, where she also directs the Center for Gynepathology Research. She is a 2006 recipient of a MacArthur Fellowship, commonly referred to as the "MacArthur genius award."
Life.
She was raised in Decatur, Valdosta, and Roswell, Georgia. She graduated with a B.ChE. in 1982 from the Georgia Institute of Technology, where she was a writer and editor on the undergraduate newspaper, "The Technique", in 1982, and was named a distinguished alumna of her alma mater's School of Engineering in 2006. She received a PhD in chemical engineering from the University of California, Berkeley in 1988. She joined the MIT faculty in 1991, was promoted to associate professor of chemical engineering in 1996, and to tenure in chemical engineering in 1998, the same year she joined the newly formed Division of Biological Engineering and Environmental Health at MIT. As an assistant professor, she joined a collaboration with Charles Vacanti and Joseph Upton to create tissue engineered cartilage in the shape of a human ear (published under the surname used in her first marriage), known as the Vacanti mouse. The Griffith Lab at MIT currently focuses on molecular biomaterials and tissue engineering approaches for regenerative medicine, drug development and understanding disease pathophysiology.
In 1994, together with colleagues Roger Kamm and Alan Grodzinsky, she led development of MIT's first interdepartmental minor degree, in biomedical engineering, which was launched in 1995 and soon became MIT's most popular minor degree. The interdepartmental bioengineering curriculum committee she chaired grew into the Undergraduate Programs Committee for the Department of Biological Engineering, and as chair of this committee she led development of the undergraduate major in Biological Engineering, launched in 2005 as MIT's first new undergraduate major in 39 years. She stepped down as chair of this committee in 2009 to spend a fellowship year at the Radcliffe Institute for Advanced Study, sponsored by the Harvard Stem Cell Institute.
She currently holds the School of Engineering Teaching Innovation Chair in recognition of her contributions to curriculum development at MIT. Furthermore, she is the co-founder of CN Bio Innovations and serves on the advisory board of Lumicell. She is married to Doug Lauffenburger, also a professor at MIT.
Women's health research.
Griffith currently directs the Center for Gynepathology Research (CGR) at MIT, which she launched in 2009 together with Keith Isaacson, Director of the Newton-Wellesley Hospital Center for Minimally Invasive Gynecology Surgery. The public launch featured a passionate talk by the celebrity host of Bravo's Top Chef, Padma Lakshmi, who suffered for over a decade with endometriosis before being diagnosed. Lakshmi co-founded the Endometriosis Foundation of America (EFA) to raise awareness of the disease, especially among college students. Lakshmi's experience underscored that of Griffith's own 16-year-old niece, who was diagnosed with endometriosis after suffering years of debilitating pain, which had been attributed to "stress" instead of to a treatable disease. The average delay between onset of symptoms and diagnosis of endometriosis is about ten years; Griffith's niece was diagnosed with less delay only because a family member with the disease insisted she see a gynecological surgeon who specializes in treating endometriosis. The CGR now has over 10 participating faculty at MIT and collaborates with surgeons and scientists in Brazil, Singapore, and across the US. Griffith was honored at the EFA's annual Blossom Ball in NYC in 2010 for her efforts to raise awareness about endometriosis among scientists and engineers.


L-pipecolate dehydrogenase

In enzymology, a L-pipecolate dehydrogenase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are L-pipecolate and acceptor, whereas its two products are 2,3,4,5-tetrahydropyridine-2-carboxylate and reduced acceptor.
This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH-NH group of donors with other acceptors. The systematic name of this enzyme class is L-pipecolate:acceptor 1,6-oxidoreductase. This enzyme is also called L-pipecolate:(acceptor) 1,6-oxidoreductase. This enzyme participates in lysine degradation.


Protein-disulfide reductase (glutathione)

In enzymology, a protein-disulfide reductase (glutathione) () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are glutathione and protein disulfide, whereas its two products are glutathione disulfide and protein dithiol.
This enzyme belongs to the family of oxidoreductases, specifically those acting on a sulfur group of donors with a disulfide as acceptor. The systematic name of this enzyme class is glutathione:protein-disulfide oxidoreductase. Other names in common use include glutathione-insulin transhydrogenase, insulin reductase, reductase, protein disulfide (glutathione), protein disulfide transhydrogenase, glutathione-protein disulfide oxidoreductase, protein disulfide reductase (glutathione), GSH-insulin transhydrogenase, protein-disulfide interchange enzyme, protein-disulfide isomerase/oxidoreductase, thiol:protein-disulfide oxidoreductase, and thiol-protein disulphide oxidoreductase. This enzyme participates in glutathione metabolism.
Structural studies.
As of late 2007, only one structure has been solved for this class of enzymes, with the PDB accession code .


6C RNA

6C RNA is a class of non-coding RNA present in actinomycetes. 6C RNA was originally discovered as a conserved RNA structure having two stem-loops each containing six or more cytosine (C) residues. Later work revealed that 6C RNAs in "Streptomyces coelicolor" and "Streptomyces avermitilis" have predicted rho-independent transcription terminators, and microarray and reverse-transcriptase PCR experiments indicate that the "S. coelicolor" version is transcribed as RNA. Transcription of the "S. coelicolor" RNA increases during sporulation, and three transcripts were detected that overlap the 6C motif, but have different apparent start and stop sites.
Additional work established that 6C RNAs regulate a variety of protein-coding genes by acting as "trans"-acting antisense RNAs. Among the genes regulated by 6C RNAs, many are involved in DNA replication and the export of proteins. Bacteria in other studied phyla such as Proteobacteria and Firmicutes use proteins such as Hfq that facilitate antisense RNA interactions, but 6C RNAs do not appear to depend on homologs of Hfq or other proteins. This lack of protein dependence might be typical of Gram-positive bacteria whose genomes have high GC-content, of which actinomycetes are a prominent example.


Dyadobacter arcticus

Dyadobacter arcticus is a Gram-negative, psychrotolerant, rod-shaped, aerobic and non-motile bacterium from the genus of Dyadobacter which has been isolated from Arctic soil from Svalbard Archipelago in Norway.


Jan-Hendrik S. Hofmeyr

Jan-Hendrik Hofmeyr was born in Durban, South Africa on 25th August 1953. He is one of the leaders in the field of metabolic control analysis and the quantitative analysis of metabolic regulation.
Education.
Hofmeyr obtained BSc hons. (1976), MSc (1978) and PhD (1986) at the University of Stellenbosch. While preparing his doctoral thesis he spent six months with Athel Cornish-Bowden at Birmingham and three months with Henrik Kacser at Edinburgh. Both of these visits led to long-term collaborations.
Research.
Hofmeyr's doctoral research concerned the use of graphical patterns to elucidate chains of interaction in metabolic regulation, later published in the European Journal of Biochemistry, and his collaboration with Kacser led to a study of the effect of moiety-conservation on control of pathways. At this time he and Cornish-Bowden were concerned that the development of metabolic control analysis seemed to be almost independent of the knowledge of metabolic regulation that had grown from the recognition of regulatory mechanisms in the 1950s and 1960s, most notably the importance of feedback inhibition and cooperative behaviour of enzymes. This led them to propose a way of quantifying metabolic regulation, the first of a series of publications that culminated in an analysis of the role of supply and demand in biochemical systems, i.e. an analysis of how negative feedback allow metabolic pathways to respond to changes in the demand for metabolites while resisting variations in the supply of starting materials.
During the 21st century Hofmeyr has applied ideas of control analysis to ecosystems, and to the understanding of the self-organization of cell function in the spirit of Robert Rosen. More recently he has worked on the development of code biology, the novel discipline founded by Marcello Barbieri that recognizes that the genetic code is just one of several codes used and needed by biological systems.
Career.
Appointed as Junior Lecturer in the Biochemistry Department of the University of Stellenbosch in 1975, Hofmeyr eventually became Distinguished Professor in 2014, and Emeritus Professor in 2019. Between 2009 and 2015 he was Co-director and then Director of the Centre for Studies in Complexity at Stellenbosch, which he had co-founded in 2009.
Prizes and distinctions.
Harry Oppenheimer Fellowship Award and Gold Medal (2002)

Beckman Coulter Gold Medal of the South African Society for Biochemistry and Molecular Biology (2003)

Havenga prize for Biological Sciences from the Suid-Afrikaanse
Akademie vir Wetenskap en Kuns (South African Academy for
Science and Art) (2009)
Other activities.
In addition to his scientific research, Hofmeyr is a classically trained flute player
and also plays the baroque flute, guitar and banjo.
He was one of the composers and performers who helped launch the Afrikaans "Kabaret"
tradition in the 1980s in South Africa, through his work with authors, composers and directors.
His classic scores for lyrics of Hennie Aucamp and Etienne van Heerden have become standard items in Afrikaans popular
music. He has also played older characters in productions of the University of Stellenbosch Drama Department.


3Scan

3Scan, Inc. was an American biotechnology company based in San Francisco, California. It offered automated microscopy services using a coordinated combination of both hardware and software for the 3D analysis of cells, tissues, and organs. The company was founded in 2011 by Todd Huffman, Megan Klimen, Matthew Goodman, and Cody Daniel. The 3Scan technology is based on the Knife Edge Scanning Microscope developed in the late 1990s by Bruce McCormick, founder of the Brain Networks Lab at Texas A&M University.
History.
3Scan CEO Todd Huffman originally worked as a neuroinformatics researcher at Texas A&M in 2003 and first encountered the technology which became the core of 3Scan's microscopy services during this time. While the KESM was originally developed as a neuroimaging tool, 3Scan has taken the principles involved in this technology and expanded its use to create a novel type of histology and tissue imaging.
The company has raised a total of $22 million through two rounds of equity funding from Lux Capital and Data Collective, joined by Dolby Family Ventures, OS Fund, Comet Labs, and Breakout Ventures. They have also raised $390,000 in non-dilutive grant money from institutions such as Breakout Labs and Start-Up Chile. The company employs about 35 full-time employees and generates revenue by providing its imaging services to pharmaceutical companies, labs, or pathologists involved in pre-clinical drug discovery. As of 2017, 3Scan is collaborating with pharmaceutical development scientists, academic researchers, and tissue engineers as a contract research organization. It is not currently used for clinical diagnostics, but is in the process of building out a platform to perform clinical pathology and diagnostic services.
In 2018, cofounder Cody Daniel was named on the "Forbes" 30 Under 30 list.
In 2019, 3Scan became a part of Strateos.
Technology.
3Scan’s technology changes the histology workflow to automate the process of tissue sectioning and imaging. Traditionally, histology slides are prepared by embedding a tissue sample in paraffin, taking some few slices of the tissue with a hand-driven microtome, mounting the tissue slices on slides, and staining with various histology stains. A pathologist then looks at these slides with a microscope. In the workflow developed by 3Scan, tissues are stained, embedded, and then sliced and imaged by the KESM. By automating this process, it becomes feasible to take slices of and image the entirety of tissue instead of a few representative slides. In addition to taking more sections, 3Scan’s proprietary software is able to computationally realign these sections in order to visualize the 3D structure of the tissue, similar in appearance to a CT scan. It uses machine vision to produce a 3D spatial map that researchers can analyze on a screen, similar to a topographical map of the Earth. Using these maps, scientists are also able to analyze parts of tissue and phenomena that were previously not visible, such as changes to blood flow between different tissue samples.
This analysis is not possible using traditional manual histology techniques due to the excessive time involved and lost sections due to folding and warping. 3Scan’s KESM technology is able to perform approximately a year’s worth of tissue sample processing in a single day.


Henry Caesar

Sir Henry Caesar (2 October 1630 – 6 January 1668 ) was an English politician who sat in the House of Commons in 1660 and 1666 through 1668.
Caesar was the son of Sir Charles Caesar, by his wife Jane Barkham, and succeeded to the estate of Bennington, Hertfordshire, in 1642. He was admitted at Jesus College, Cambridge, on 10 June 1646 and admitted at Inner Temple in 1647.
In 1660, Caesar was elected Member of Parliament for Hertfordshire in the Convention Parliament. He regained his seat in Cavalier Parliament in a by-election in April 1666. He was knighted on 7 July 1660.
Caesar married Elizabeth (1630 - 1670) daughter of Robert Angell a merchant of London by his wife Susan Bateman, on 6 November 1649 at St Olave, Hart Street, London.
Caesar died of smallpox in 1668 at the age of 37. He was survived by his son Charles who became a member of parliament and his daughter Jane wife of Sir Thomas Pope Blount, 1st Baronet.


OverBlood

 is a science fiction video game developed by Riverhillsoft and published by Electronic Arts for the PlayStation in 1997. It is considered the first survival horror game to make use of a fully three-dimensional virtual environment. The game was released on the Japanese PlayStation Network Store on February 23, 2011.
Gameplay.
Part adventure game and part survival horror, "Overblood" incorporates elements of arcade, fighting and puzzle games. The player is able to toggle the camera between first- and third-person, both of which are required to solve the game's various puzzles.
Synopsis.
Plot.
"Overblood" takes place at Lystra Laboratories' hidden research center where a team of scientists have been conducting controversial genetic experiments. The game begins when a system malfunction releases the player character, Raz Karcy (Lars in European releases), from a cryogenic container. Cold and confused, he awakens with no memory. Concerns about his identity are soon replaced by an urgent need to escape, as he reveals the scientists' fateful plan and his role in it.
Characters.
The game features three playable characters. The majority of the game is played as Raz Karcy, the game's protagonist. Other playable characters include Milly Azray, a woman who befriends Raz, and Pipo, a small and very helpful robot.
Reception and legacy.
Reviews for "Overblood" were mixed, with critics generally commenting that it has enough interesting elements to make it respectable, but is ultimately not worth buying. The most frequent criticism was that the game is too slow-paced and lacking in tension, with too much time spent wandering corridors and opening doors, relatively little time spent on puzzles, and far too little time spent on action sequences. Compounding this problem, many critics found the action sequences too simplistic. Joe Fielder elaborated in "GameSpot" that "They entail moving at just the right moment and are more timing than skill-based."
The need to switch between three different playable characters in order to solve certain puzzles was often cited as one of "OverBlood"s most interesting aspects. Some also praised the story. The game was reviewed by "GameFan Magazine", where the game's previously rumored similarities to "Resident Evil" were dismissed and strong comparisons drawn instead to "Doctor Hauzer" for which "Overblood" (created by the same development team) serves as a spiritual successor. The game was criticized for the appearance of the characters, their unrealistic movements, and the overall plot, and the game received low marks for play mechanics and controls. IGN stated, ""OverBlood" attempts to recreate the moodiness and horror of "Resident Evil" but never quite succeeds." Crispin Boyer of "Electronic Gaming Monthly" made the same unfavorable comparison to "Resident Evil", but concluded more positively: "That's OK, because "Overblood" is still an engrossing adventure ... that packs a solid mix of puzzles and exploration." "Next Generation", by contradiction, stated that "Despite the excellent graphics, the game just can't hold a player's interest. "OverBlood" could've used a little more action and a lot less wandering around."
German magazine "Maniac" gave it a score of 68 out of 100.
In 1998, a direct sequel was released entitled "Overblood 2".


Ilona Banga

Ilona Banga (1906–1998) was a Hungarian biochemist known for co-discovering actomyosin and working to characterize how actin and myosin interact to produce muscle contraction. She and her husband József Mátyás Baló discovered the first elastase – an enzyme capable of degrading the protein elastin which gives tissues like veins their flexibility. She also contributed to work that earned Albert Szent-Györgyi the Nobel Prize in Physiology or Medicine in 1937, including by developing methods for the purification and characterization of large quantities of vitamin C. During World War II she saved the equipment of the Institute of Chemistry of the University of Szeged.
Early life and education.
Banga was born February 3, 1906, in the southeastern Hungarian town of Hódmezővásárhely. She was interested in becoming a medical doctor but chose instead to study chemistry because her mother didn't think medical doctor was a proper profession for a woman.
She started studies in Szeged, continued at the University of Vienna and received an MSc in chemistry from the University of Debrecen in 1929. At the University of Debrecen she carried out Physiology research under the guidance of professor Fritz Verzár.
Career.
After graduating, she joined the laboratory of Albert Szent-Györgyi at the University of Szeged's Institute for Medicinal Chemistry as a research assistant – she was the first associate for this future Nobel laureate. Banga worked with Szent-Györgyi for almost fifteen years, resulting in 25 joint publications. Banga later spent time working abroad in Liege, Belgium, and Oxford, England. While in Oxford she worked with another future Nobel laureate, Severo Ochoa to study vitamin B1.
In 1945, Albert Szent-Györgyi moved his lab from Szeged to Budapest and Banga followed him there. She later became chief of the Chemical Laboratory of the First Institute of Pathological Anatomy in Budapest, where she studied arteriosclerosis and aging with her husband József Baló. She retired in 1970. After retiring she remained engaged in the scientific community, serving as a scientific advisor to the Gerontology Institute from 1971 to 1986.
Banga was never made a professor, even though (in 1950) she received her DSc degree, making her eligible. She was, however, the first woman to achieve the rank of Privatdozent (comparable to associate professor) at the University of Szeged (1940). She authored two scientific texts, including "Structure and Function of Elastin and Collagen", and was a founder member of the Hungarian Biochemical Society.
Research.
Banga's initial work as Szent-Györgyi's associate involved studying carbohydrate metabolism. She developed methods for the large-scale purification of ascorbic acid (vitamin C) from Hungarian paprika – work that entailed extracting the vitamin from close to a metric ton of paprika. After winning the Nobel Prize in Physiology or Medicine in 1937 (for work to which Banga contributed), Albert Szent-Györgyi switched his lab's focus to researching muscle contraction, motivated by the findings of Engelhardt and Ljubimova that the muscle protein myosin wasn't merely a structural protein – it had phosphatase (ATPase) activity. Banga confirmed these findings and further characterized myosin's ability to split ATP. In the process, she co-discovered actomyosin.
In order to get myosin to analyze, Banga extracted it from rabbit muscles – she minced rabbit muscles and extracted myosin from them following an established high salt protocol. One time she ran out of time to do the extraction and left the minced muscle sitting in saline overnight and when she came back the next morning it had changed from its usual thin liquid appearance to a thick, viscous solution. They found that if they added ATP to it the viscosity would decrease. Further work by another lab member, Brunó Straub, showed that Banga had extracted a combination of actin and myosin, which they called actinomyosin. Straub was able to isolate the actin and he, Banga, and other members of the lab carried out extensive characterization of actin, myosin, and actinomyosin, showing that it is responsible for muscle contraction.
The widespread distribution of their findings was hampered by World War II – Szent-Györgyi was wanted for anti-Nazi activities and went into hiding, and other male colleagues left the lab to escape drafting. Banga remained behind and saved the Department of Medicinal Chemistry and its equipment by posting signs on the door in German, Hungarian, and Russian identifying the facilities as an infectious disease laboratory and reporting hours for the drop-off of infectious materials. These signs warded off would-be thieves and made the Department of Medicinal Library the only institution at the University to come out of WWII with all of its equipment and facilities preserved.
Albert Szent-Györgyi left Hungary for the United States after the war, but Ilona stayed and became chief of the Chemical Laboratory of the First Institute of Pathological Anatomy in Budapest. The new position also came with a shift in focus – she became deeply involved in gerontology, the study of aging, with a specific interest in how the connective tissue of blood vessel walls changes with age. She carried out seminal work on arteriosclerosis with her husband József Mátyás Baló. One area of their investigation determining what causes degradation of the elastin fibers holding together the walls of veins. They discovered an enzyme made by the pancreas could degrade elastin and named it elastase. This is now known to be only one of a class of peptidases capable of cleaving elastin, and the pancreatic elastase Banga and Baló found is unlikely to play a role in arteriosclerosis, but it was the first elastolytic enzyme discovered. The discovery was met with skepticism initially, but, by crystallizing elastase, Banga was able to clear up doubt. Banga further characterized this enzyme and published more than 60 articles on elastase, elastin, and related molecules and processes during the period from 1948 to 1965.
Personal life.
Banga married the pathologist József Mátyás Baló in Szeged in 1945, working closely with him on research on arteriosclerosis until his death in 1979. The couple had a son, Mátyás Jr., who became an academic dermatologist. Banga died March 11, 1998.
Honors and awards.
Banga was awarded the Kossuth Prize in 1955 for her discovery of elastase – it was offered to her in 1952 but in what's believed to be an oversight, her husband and co-discoverer József Mátyás Baló was not included so, responding that the work was a joint effort, she declined the offer, accepting it in 1955 when the offer was made to the pair of them. She was the first awardee of the University of Szeged's Albert Szent-Györgyi Commemorative Medal (1986) and was elected to the Leopoldina Academy in Halle, East Germany in 1962.


6-Chloronicotine

6-Chloronicotine is a drug which acts as an agonist at neural nicotinic acetylcholine receptors. It substitutes for nicotine in animal studies with around twice the potency, and shows antinociceptive effects.


Chrysosporide

Chrysosporide is a cyclic pentapeptide. It is isolated from the mycoparasitic fungus "Sepedonium chrysospermum", found in "New Zealand".


Ribonucleoprotein particle

A ribonucleoprotein particle or RNP is a complex formed between RNA and RNA-binding proteins (RBPs). The term RNP foci can also be used to denote intracellular compartments involved in processing of RNA transcripts.
RNA/RBP complexes.
RBPs interact with RNA through various structural motifs. Aromatic amino acid residues in RNA-binding proteins result in stacking interactions with RNA. Lysine residues in the helical portion of RNA binding proteins help to stabilize interactions with other nucleic acids as a result of the force of attraction between the positively-charged lysine side chains and the negatively-charged phosphate "backbone" of RNA.
It is hypothesized that RNA sequences in the 3'-untranslated region determine the binding of RBPs, and that these RBPs determine the post-transcriptional fate of mRNAs.
RNP granules.
RNP granules are a highly diverse group of compartments. These include stress granules, processing bodies, and exosomes in somatic cells. Many RNP granules are cell type and/or species specific. For example, chromatoid bodies are found only in male germ cells, whereas transport granules have so far been found only in neurons and oocytes. RNP granules function mainly by physically separating or associating transcripts with proteins. They function in the storage, processing, degradation and transportation of their associated transcripts.
RNP granules have been shown to have particular importance in cells where post-transcriptional regulation is of vital importance. For example, in neurons where transcripts must be transported and stored in dendrites for the formation and strengthening of connections, in oocytes/embryos where mRNAs are stored for years before being translated, and in developing sperm cells where transcription is halted before development is complete.


Difference gel electrophoresis

Difference gel electrophoresis (DIGE) is a form of gel electrophoresis where up to three different protein samples can be labeled with size-matched, charge-matched spectrally resolvable fluorescent dyes (for example Cy3, Cy5, Cy2) prior to two dimensional gel electrophoresis.
Procedure.
The three samples are mixed and loaded onto IEF (isolectric focusing chromatography) for first dimension and the strip is transferred to a SDS PAGE. After the gel electrophoresis, the gel is scanned with the excitation wavelength of each dye one after the other, so each sample can be seen separately (if we scan the gel at the excitation wavelength of the Cy3 dye, we will see in the gel only the sample that was labeled with that dye). This technique is used to see changes in protein abundance (for example, between a sample of a healthy person and a sample of a person with disease), post-translational modifications, truncations and any modification that might change the size or isoelectric point of proteins. The binary shifts might be left to right (change in isoelectric point), vertical (change in size) or diagonal (change in both size and isoelectric point). Reciprocal Labeling is done to make sure the changes seen are not due to dye-dependent interactions.
Advantages.
It overcomes limitations in traditional 2D electrophoresis that are due to inter-gel variation. This can be considerable even with identical samples. Since the proteins from the different sample types (e.g. healthy/diseased, virulent/non-virulent) are run on the same gel they can be directly compared. To do this with traditional 2D electrophoresis requires large numbers of time-consuming repeats.
Standards.
In experiments comprising several gels, a common technique is to include an internal standard in each gel. The internal standard is prepared by mixing together several or all of the samples in the experiment. This allows the measurement of the abundance of a protein in each sample relative to the internal standard. Since the amounts of each protein in the internal standard is known to be the same in every gel, this method reduces inter-gel variation.electrophoresis incorporating a pooled internal standard.


Prucalopride

Prucalopride, brand name Prudac, among others, is a drug acting as a selective, high affinity 5-HT4 receptor agonist which targets the impaired motility associated with chronic constipation, thus normalizing bowel movements. Prucalopride was approved for medical use in the European Union in 2009, in Canada in 2011, in Israel in 2014, and in the United States in December 2018. The drug has also been tested for the treatment of chronic intestinal pseudo-obstruction.
Medical uses.
The primary measure of efficacy in the clinical trials is three or more spontaneous complete bowel movements per week; a secondary measure is an increase of at least one complete spontaneous bowel movement per week. Further measures are improvements in PAC-QOL (a quality of life measure) and PAC-SYM (a range of stool, abdominal, and rectal symptoms associated with chronic constipation). Infrequent bowel movements, bloating, straining, abdominal pain, and defecation urge with inability to evacuate can be severe symptoms, significantly affecting quality of life.
In three large clinical trials, 12 weeks of treatment with prucalopride 2 and 4 mg/day resulted in a significantly higher proportion of patients reaching the primary efficacy endpoint of an average of ≥3 spontaneous complete bowel movements than with placebo. There was also significantly improved bowel habit and associated symptoms, patient satisfaction with bowel habit and treatment, and HR-QOL in patients with severe chronic constipation, including those who did not experience adequate relief with prior therapies (>80% of the trial participants). The improvement in patient satisfaction with bowel habit and treatment was maintained during treatment for up to 24 months; prucalopride therapy was generally well tolerated.
Contraindications.
Prucalopride is contraindicated where there is hypersensitivity to the active substance or to any of the excipients, renal impairment requiring dialysis, intestinal perforation or obstruction due to structural or functional disorder of the gut wall, obstructive ileus, severe inflammatory conditions of the intestinal tract, such as Crohn's disease, and ulcerative colitis and toxic megacolon/megarectum.
Side effects.
Prucalopride has been given orally to ~2700 patients with chronic constipation in controlled clinical trials. The most frequently reported side effects are headache and gastrointestinal symptoms (abdominal pain, nausea or diarrhea). Such reactions occur predominantly at the start of therapy and usually disappear within a few days with continued treatment.
Mechanism of action.
Prucalopride, a first in class dihydro-benzofuran-carboxamide, is a selective, high affinity serotonin (5-HT4) receptor agonist with enterokinetic activities. Prucalopride alters colonic motility patterns via serotonin 5-HT4 receptor stimulation: it stimulates colonic mass movements, which provide the main propulsive force for defecation.
The observed effects are exerted via highly selective action on 5-HT4 receptors: prucalopride has >150-fold higher affinity for 5-HT4 receptors than for other receptors. Prucalopride differs from other 5-HT4 agonists such as tegaserod and cisapride, which at therapeutic concentrations also interact with other receptors (5-HT1B/D and the cardiac human ether-a-go-go K+ or hERG channel respectively) and this may account for the adverse cardiovascular events that have resulted in the restricted availability of these drugs. Clinical trials evaluating the effect of prucalopride on QT interval and related adverse events have not demonstrated significant differences compared with placebo.
Pharmacokinetics.
Prucalopride is rapidly absorbed (Cmax attained 2–3 hours after single 2 mg oral dose) and is extensively distributed. Metabolism is not the major route of elimination. "In vitro", human liver metabolism is very slow and only minor amounts of metabolites are found. A large fraction of the active substance is excreted unchanged (about 60% of the administered dose in urine and at least 6% in feces). Renal excretion of unchanged prucalopride involves both passive filtration and active secretion. Plasma clearance averages 317 ml/min, terminal half-life is 24–30 hours, and steady-state is reached within 3–4 days. On once daily treatment with 2 mg prucalopride, steady-state plasma concentrations fluctuate between trough and peak values of 2.5 and 7 ng/ml, respectively.
"In vitro" data indicate that prucalopride has a low interaction potential, and therapeutic concentrations of prucalopride are not expected to affect the CYP-mediated metabolism of co-medicated medicinal products.
Approval.
In the European Economic Area, prucalopride was originally approved for the symptomatic treatment of chronic constipation in women in whom laxatives fail to provide adequate relief. Subsequently, it has been approved by the European Commission for use in adults – that is, including male patients – for the same indication.


NDTDI

NDTDI is a tricyclic tryptamine derivative which is thought to act as a serotonin receptor agonist, though its pharmacology has not been studied in detail. It is a structurally simplified analogue of LSD and is reported to retain similar effects, though with many times lower potency. It has been sold as a designer drug since 2016 and was first identified by a forensic laboratory in Slovenia in 2017.
Legality.
NDTDI was made illegal in Latvia in March 2017.


Albert D. Friesen

Albert "Bert" D. Friesen, (born May 19, 1947) is a Canadian involved in biotechnology, innovation. and community service. His biotechnology career began as the first full-time employee and became the President CEO of The Winnipeg Rh Institute where he led the development of WinRho, one of Canada's first successful biotech products. He has also started numerous other biotech companies including a life science accelerator.
Friesen has been actively involved in a variety of community interests; Board of Directors Chair of Mennonite Economic Development Associates; an Executive member of the Premier's Economic Advisory Council and a member of the board of the Canadian Foundation for Innovation (CFI) chairing the Finance and Audit Committee. He has led a Linden Woods senior housing initiative, Winnipeg Mennonite Seniors Care Inc. serving as Board Chair for thirteen years and led the development of an innovative personal care home, Bridgwater Mennonite Personal Care Home, a subsidiary of the Winnipeg Mennonite Seniors Care Inc.; Founding Director and Chair, the Canadian Industry Biotech Association, known as BIOTECanada; Friesen presently serves as Board Chair and CEO of Medicure Inc.
Early life.
Friesen was born in the small village of Kane, Manitoba, Canada in a Mennonite farm community, where he was nurtured in his faith and love of the farm and science.
Biotech career.
Friesen began his biotech career as the first full-time employee, May 1971, of The Winnipeg Rh Institute becoming the President and CEO, where he led the development of WinRho. In 1984 he started ABI Biotechnology Inc., one of the first biotech companies in Canada and the first to become profitable, 1990. The Winnipeg Rh Institute and ABI Biotechnology were merged in 1990. In 1991 The Winnipeg Rh Institute and ABI Biotechnology Inc. became under the control of Apotex Inc., and subsequently the 2 organizations became Cangene Corporation and Apotex Fermentation Inc.
In 1991, Friesen started Novopharm Biotech (became Viventia Biotech), as a division of Novopharm Ltd., and Genesys Pharma Inc. Other biotech companies started were, Genesys Venture Inc., Mantex Biotech (acquired by Myogen), DiaMedica Inc., Miraculins Inc., Kane Biotech Inc., CanAm Bioresearch Inc., Medicure Inc., GVI Clinical Development Solutions Inc., Marsala Biotech Inc., Koff Pharma Inc., Waverley Pharma Inc., etc.
Other biotech activities, include being founding Board Chair, in 1988, of the biotech association in Canada, now known as BIOTECanada.
Community service.
Friesen is actively involved in a variety of community interests; an Executive member of the Premier's Economic Advisory Council and a member of the Board of Directors Canadian Foundation for Innovation Board (CFI) where he started in 1997. Was founding Board Chair of the Industrial Association of Biotech Canada (now known as BIOTECanada); served as a Director on the Advisory Board on SMEs to the Canadian Minister of Foreign Affairs and International Trade; served a member of the Industrial Advisory Committee to the Biotechnology Research Institute in Montreal. He has been a member of the Scientific Advisory Committee of the Council of Canadian Academies; Co-Chair for the Manitoba Innovation Council, Friesen has an extensive network within the international biotechnology and corporate finance communities. He has led the Lindenwoods Senior Housing initiative for thirteen years as past Board Chair of the Winnipeg Mennonite Senior Care Inc.
A strong advocate for "Business Solutions to Poverty", Friesen also chaired the international board of MEDA (Mennonite Economic Development Associates) from 2014 to 2017.
He is active in his Church and the father of three daughters and has nine grandchildren.


Janice Limson

Janice Leigh Limson is a South African Professor of Biotechnology, former Chairperson the School of Biotechnology at Rhodes University and the SARChI Chair in Biotechnology Innovation & Engagement at Rhodes University. She is founder and editor-in-chief of the magazine "Science in Africa", the first popular online science magazine for Africa. Her research focuses on topics ranging from the development of nanotechnology biosensors for cancer diagnostics, drug delivery, detection of pathogens in food to the design of fuel cell technology.
Education and career.
Limson grew up in Port Elizabeth, South Africa. She subsequently moved to Grahamstown, and she studied at Rhodes University where she completed her BSc (Honours) in 1994. In 1997 she was awarded a PhD in Chemistry at Rhodes University. She completed a Postgraduate diploma in Tertiary education from 2006 to 2008.
In 1999 Limson was a postdoctoral research fellow at the University of Texas Health Science Center at San Antonio. The next year she founded the online magazine "Science in Africa" and became its editor in chief. The magazine aims to serve "as a platform for scientists in a broad range of fields in Africa to report on their research, giving their expert views on topical issues in science and providing the right information to an African public seeking a deeper understanding of science and of the dynamic role it plays in everyone's lives."
In 2003 Limson established the BioSENS (Sensors, Energy and Nanomaterials) Research Group at Rhodes for the development of nanotechnology biosensors for cancer diagnostics and drug delivery. The establishment in 2007 of the DST/Mintek-sponsored Nanotechnology Innovation Centre at the University of the Western Cape, University of Johannesburg and Rhodes University greatly assisted this research.
From 2003 until 2009 Limson was a lecturer at Rhodes university, and in 2010 she was appointed Associate Professor at Rhodes. She also received the Vice-Chancellor's Distinguished Research Award in that year for "the volume and impact of her research outputs". In 2011 she became Chairperson of the Rhodes University School of Biotechnology. In 2014 she was awarded Professor of Biotechnology and was appointed Director of Rhodes University Biotechnology Innovation Centre. The Centre has since launched its first products, catalysts for sensors, and is working on sensors for CD4 and malaria in a partnership with UNICEF and the South African Medical Research Council.
In 2015 she was appointed to the DSI/NRF South African Research Chairs Initiative (SARChI) Chair in Biotechnology Innovation & Engagement at Rhodes.
In March 2019 she contributed to a series of public lectures on "Truth and Trustworthiness in Science" co-ordinated by the ASSAf and the British High Commission in South Africa. The discussion she was involved with was entitled: "Misunderstandings and Misuses: Science Journalism".
Limson is the editor of the South African Agency for Science and Technology Advancement (SAASTA) Nanotechnology Public Engagement Programme’s newsletter, called "Nanotech Public Engagement", and is a fellow of The African Academy of Sciences.
Awards and recognition.
Limson has received numerous awards, including:


5-MeO-MPMI

5-MeO-MPMI (also known as 5-Methoxy-"N"-methyl-(α,"N"-trimethylene)tryptamine) is a tryptamine derivative that is a psychedelic drug. It was first developed by the team led by JE Macor in 1992, and subsequently investigated by the team led by David Nichols from Purdue University in the late 1990s. This compound produces psychedelic-appropriate responding in animal tests with a similar potency to the amphetamine-derived psychedelic DOI, and has two enantiomers, with only the ("R")-enantiomer being active.


4-Fluoro-5-methoxy-DMT

4-Fluoro-5-Methoxy-"N","N"-dimethyltryptamine (4-F-5-MeO-DMT) was first described by David E. Nichols team in 2000. It is a potent 5-HT1A agonist. Substitution with the 4-fluorine markedly increased 5-HT1A selectivity over 5-HT2A/2C receptors with potency greater than that of the 5-HT1A agonist 8-OH-DPAT.
The analog compound with the "N","N"-dialkyl substituents constrained into a pyrrolidine ring, is a slightly stronger agonist for the 5-HT1A receptor and retains the selectivity over the 5-HT2A/2C receptors.


5,6-MDO-DiPT

5,6-MDO-DiPT, or 5,6-methylenedioxy-"N","N"-diisopropyltryptamine, is a lesser-known psychedelic drug. It is the 5,6-methylenedioxy analog of DiPT. 5,6-MDO-DiPT was first synthesized by Alexander Shulgin. It is mentioned in his book "TiHKAL" ("Tryptamines I Have Known and Loved"), but 5,6-MDO-DiPT was never tested to determine what effects it produces, if any. Very little data exists about the pharmacological properties, metabolism, and toxicity of 5,6-MDO-DiPT.


ELISpot

The enzyme-linked immune absorbent spot (ELISpot) is a type of assay that focuses on quantitatively measuring the frequency of cytokine secretion for a single cell. The ELISpot Assay is also a form of immunostaining since it is classified as a technique that uses antibodies to detect a protein analyte, with the word analyte referring to any biological or chemical substance being identified or measured.
The FluoroSpot Assay is a variation of the ELISpot assay. The FluoroSpot Assay uses fluorescence in order to analyze multiple analytes, meaning it can detect the secretion of more than one type of protein.
History.
Cecil Czerkinsky first described ELISpot in 1983 as a new way to quantify the production of an antigen-specific immunoglobulin by hybridoma cells.
In 1988, Czerkinsky developed an ELISA spot assay that quantified the secretion of a lymphokine by T cells.
In the same year, dual-color ELISpot was combined with computer imaging for the first time, which allowed for the enumeration and analysis of spots.
1988 also marked the first use of membrane-bottomed plates for performing these assays.
Mechanism of ELISpot.
Mechanism of FluoroSpot.
The FluoroSpot assay is very similar to the ELISpot assay. The main difference is that the FluoroSpot assay is able to analyze the presence of multiple analytes on one plate of wells, whereas the ELISpot assay can only analyze one analyte at a time. The FluoroSpot assay accomplishes this by using fluorescence rather than an enzymatic reaction for detection. The steps for a FluoroSpot assay are also similar, with a few differences.
Since the FluoroSpot assay identifies and quantifies the presence of multiple analytes, it is possible that the absorption of one analyte can affect the secretion of another analyte; this is called capture effects. The affect an analyte has on another analyte could be positive or negative (the production of the second analyte can either increase or decrease). To counteract capture effects, it is possible to use co-stimulation in order to bypass the decreased production of an analyte. This is when a second antibody that stimulates the production of the same analyte is added to the wells.
Applications of ELISpot and FluoroSpot.
The ELISpot and FluoroSpot assays can be used in many research fields: vaccine development, cancer, allergies, monocytes/macrophages/dendritic cells characterization, apolipoproteins analysis, and veterinary research. With the ELISpot, you can study antigen-specific cytokine responses, antibody specific secreting cells, tumor antigens, granzyme B and Perforin release by T cells, vaccine efficacy, epitope mapping, cytotoxic T-cell activity, detection of IL-4, IL-5, and IL-13, vaccine-induced antibody responses, antigen-specific memory B cells, and much more.
More specifically, the T-cell ELISpot assay is used to characterize T-cell subsets. This is because the assay can detect the production of cytokines IFN-y, IL-2, TNF-alpha, IL-4, IL-5, and IL-13. The first three cytokines are produced by Th1 cells, while the last three are produced by Th2 cells. Measuring T-cell responses through cytokine production also makes it possible to study vaccine efficacy.
With T-cell FluoroSpot, you can monitor tumor-infiltrating lymphocytes. You can also analyze the IFN-y cytokine and granzyme B secretion in order to assess cytotoxic T-cell responses. Both of these are used for cancer research.
With B-cell FluoroSpot, vaccine efficacy can also be observed by quantifying the secretion of IgG, IgA, and IgM before and after a vaccination. This analysis of multiple immunoglobulins is made possible because of the fluorescence method used in the FluoroSpot.


Alkylphosphocholine

Alkylphosphocholines are phospholipid-like molecules that have been synthesised, which have remarkable biological and therapeutic activities. They are phosphocholine esters of aliphatic long chain alcohols differing in chain length, unsaturation and position of the cis-double bond.


Carol Kumamoto

Carol Kumamoto is an American microbiologist who is Professor of Molecular Biology & Microbiology at Tufts University. She investigates the filamentous growth of "Candida albicans", a fungal pathogen that causes several diseases. She is also interested in how "C. albicans" interacts with its host during colonisation and invasive diseases. She is a Fellow of the American Association for the Advancement of Science and the American Society for Microbiology.
Early life and education.
Kumamoto studied biology at the University of California, Los Angeles (UCLA), graduating in 1976. She remained there for her doctoral degree before moving to Harvard Medical School for a postdoctoral research position in 1980. She joined Stanford University as a research associate and joined Tufts University in 1986. Her early work involved monitoring the mechanism of action of "E.coli". She studied the mutant strains of "E.Coli" that were defective in exporting the periplasmatic protein maltose binding protein into the periplasm from the cytoplasm. She showed that these mutants were defective in the gene SecB and went on to study the mechanism of action of this protein; showing that it had chaperone activity and was selective in its binding to exported protein precursors.
Research and career.
Kumamoto is a Professor of Molecular Biology & Microbiology at Tufts University. Her research considers "Candida albicans" ("C. albicans"), a fungal pathogen that can causes several diseases. "C. albicans" typically colonises the gastrointestinal tract of humans without causing any problems, but when the host organism becomes immunocompromised the pathogen produces invasive lesions that are associated with "candidiasis". "Candidiasis" is complicated to diagnose and can be fatal. Kumamoto demonstrated that low levels of the protein Efg1p permit fast growth of "C. albicans", whereas high levels suppress the growth.
In particular, Kumamoto studies the environmental conditions that cause "C. albicans" to grow in filamentous, elongated cells, which are able to invade and destroy biological tissue and enter the bloodstream. Early in her career, Kumamoto demonstrated that this filamentous growth occurs when the organism is grown in contact with an agar medium. She showed that the CZF1 gene is a regulator of the filamentation response, and that Mkc1 and Cek1 (MAP kinases) are activated when cells are grown in contact with the agar. She has demonstrated that certain dietary fats, including coconut oil, can suppress the growth of "C. albicans" in the gut, decreasing the risk of fungal infections. It has been proposed that this finding will decrease the need for antifungal drugs and could be used to decrease the amount of "C. albicans" in the guts of premature infants. In 2019, Kumamoto reported the first results of a clinical trial that involved supplementing the diets of preterm infants with medium-chain triglycerides to reduce the amount of "C. albicans" in the gastrointestinal tract.
Kumamoto has also studied "candida auris" a fungus that was identified in 2009 that appears to withstand anti-fungal drugs. She is interested in the mechanism by which "C. albicans" interacts with its host during colonisation and invasive degrees. For example, Kumamoto has demonstrated that the interaction of "C. albicans" with other pathogens can influence its virulence.
Selected publications.
Kumamoto has written for "The Conversation".


Lysine 2-monooxygenase

In enzymology, a lysine 2-monooxygenase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are L-lysine and O2, whereas its 3 products are 5-aminopentanamide, CO2, and H2O.
This enzyme belongs to the family of oxidoreductases, specifically those acting on single donors with O2 as oxidant and incorporation of two atoms of oxygen into the substrate (oxygenases). The oxygen incorporated need not be derived from O with incorporation of one atom of oxygen (internal monooxygenases o internal mixed-function oxidases). The systematic name of this enzyme class is L-lysine:oxygen 2-oxidoreductase (decarboxylating). Other names in common use include lysine oxygenase, lysine monooxygenase, and L-lysine-2-monooxygenase. This enzyme participates in lysine degradation. It employs one cofactor, FAD.


United States Army Medical Research and Development Command

The United States Army Medical Research and Development Command (USAMRDC) is the United States Army's medical materiel developer, with responsibility for medical research, development, and acquisition and medical logistics management. The USAMRDC's expertise in these critical areas helps establish and maintain the capabilities the U.S. Army needs to fight and win on the battlefield.
Overview.
USAMRDC Headquarters at Fort Detrick, Maryland, supports 11 subordinate commands located throughout the world. Six USAMRDC medical research laboratories and institutes perform the core science and technology (S&T) research to develop medical solutions. These laboratories specialize in various areas of biomedical research, including infectious diseases, combat casualty care, operational medicine, clinical and rehabilitative medicine, chemical and biological defense, combat dentistry, and laser effects. The laboratories are staffed with highly qualified scientists and support personnel.
A large extramural research program and numerous cooperative research and development (R&D) agreements provide additional S&T capabilities by the leading R&D organizations in the civilian sector.
Five USAMRDC subordinate commands perform medical materiel advanced development, strategic and operational medical logistics, and contracting, to complete the lifecycle management of medical materiel.
About 6,000 military, civilian, and contractor personnel are assigned to support the Headquarters and subordinate units. Officers, enlisted Soldiers, and civilians-many of whom are among the most respected and knowledgeable specialists in their fields-provide subject matter expertise in medical, scientific, and technical areas throughout the Command.
Medical information and products developed by USAMRDC protect and sustain the health and safety of the force through deployment and combat. The USAMRDC motto, "Protect, Project, Sustain," emphasizes the Command's priorities in support of the warfighter.
History.
In March 1994, a merger of Medical Research and Development Command, the United States Army Medical Materiel Agency and the Health Facilities Planning Agency resulted in creation of the Medical Research, Development, Acquisition and Logistics Command (MRDALC), subordinate to the then provisional MEDCOM. The MRDALC was soon renamed the U.S. Army Medical Research and Materiel Command (USAMRMC).
MRDC’s subordinate commands.
At Fort Detrick:
Elsewhere:


Valspodar

"
Valspodar (PSC833) is an experimental cancer treatment and chemosensitizer. It is a derivative of ciclosporin D (cyclosporin D).
Its primary use is as an inhibitor of the efflux transporter P-glycoprotein. Previous studies in animal models have found it to be effective at preventing cancer cell resistance to chemotherapeutics, but these findings did not translate to clinical success.
Adverse effects.
Valspodar can cause nerve damage.


CsgD

CsgD is a transcription and response regulator protein referenced to as the master modulator of bacterial biofilm development. In "E. coli" cells, CsgD is tasked with aiding the transition from planktonic cell motility to the stationary phase of biofilm formation, in response to environmental growth factors. A transcription analysis assay (happening "in vitro") illustrated a heightened decrease in CsgD's DNA-binding capacity when phosphorylated at A.A. D59 of the protein's primary sequence. Therefore, in the protein's active form (unphosphorylated), CsgD is capable of carrying out its normal functions of regulating curli proteins (fimbria) and producing ECM polysaccharides (cellulose). Following a promoter-"lacZ" fusion assay of "CsgD binding" to specific target sites on E. coli's genome, two classes of binding targets were identified: "group I genes" and "group II genes". The group I genes, akin to "fliE" and "yhbT", exhibit repressed transcription following their interaction with CsgD, whilst "group II genes", including "yccT" and "adrA", illustrated active functionality. Other group I operons that illustrate repressed transcription include "fliE" and "fliEFGH," for motile flagellum formation. Other group II genes, imperative to the transition towards stationary biofilm development, include "csgBA", encoding for curli fimbriae, and "adrA", encoding for the synthesis of cyclic diguanylate. In this context, c-di-GMP functions as a bacterial secondary messenger, enhancing the production of extracellular cellulose and impeding flagellum production and rotation.


Rev-ErbA

The Rev-ErbA proteins are members of the nuclear receptor family of intracellular transcription factors. There are two forms of the receptor, alpha and beta, each encoded by a separate gene ( and respectively).
The rev-Erb-α gene is highly unusual in that it is encoded on the opposite strand of the alpha-thyroid hormone receptor (TR) gene.
The rev-Erb-α protein is a key regulatory component of the circadian clock. In addition, rev-Erb-α appears also to regulate the breakdown of cartilage.


Zodwa Dlamini (biochemist)

 Zodwa Dlamini is a South African biochemist and Ex-deputy Vice Chancellor for Research at the Mangosuthu University of Technology. She researches molecular oncology. She is a former Vice President of the South African Medical Research Council and is a member of the Council for Scientific Advisers for the International Centre for Genetic Engineering and Biotechnology.
Early life and education.
Dlamini was born in the KwaZulu-Natal North Coast. She worked as an administration clerk in provincial health before starting a degree in Pharmacy. She was not happy, and switched to Biochemistry and Microbiology, earning Bachelor's degrees at the University of the Western Cape. She moved to the University of Natal for her Master's and PhD.
Career.
After her PhD, Dlamini returned to the University of the Western Cape as a postdoctoral fellow in molecular oncology. In 2002 she joined the University of the Witwatersrand, where she studied the toxicity of traditional African beer. In 2007 she was awarded a National Cancer Institute investigator opportunity award. She joined the University of Limpopo as an Associate Professor in 2014.
She was appointed the lead of the research directorate at the Mangosuthu University of Technology in 2015. In July 2017 Dlamini was confirmed as a member of the Council for Scientific Advisers for the International Centre for Genetic Engineering and Biotechnology. She is a Visiting Professor at the University of Bristol.
Dlamini is concerned about education within the local community and supports schools in accounting, maths and physics. She believes that Mangosuthu University of Technology can improve the lives of the people in Umlazi. She is a member of the steering committee of the Academy of Science of South Africa's STEM education group.
In 2017 she led a delegation of Mangosuthu University of Technology scientists to the Dublin Institute of Technology. She is interested in precision medicine. Dlamini has contributed to two books about cancer, including "Current Immunotherapeutic Treatments in Colon Cancer" and "South African Herbal Extracts as Potential Chemopreventive Agents: Screening for Anticancer Splicing Activity". Her current research looks at abnormal MicroRNA in cancers associated with HIV. The project received R6 million over three years from the South African Medical Research Council.


23S methyl RNA motif

The 23S methyl RNA motif is a conserved RNA structure found upstream of genes predicted to encode rRNA methyltransferases, possibly for 23S rRNA. However, in one case it is far (3 kilobases) from the rRNA methyltransferase gene. Nonetheless, it was proposed that this RNA could be a cis-regulatory element, an attractive hypothesis in view of the fact that rRNA methyltransferases can bind RNA, and therefore presumably the 23S methyl RNA. This regulatory strategy is common in bacteria to control levels of ribosomal subunits, using RNA motifs called ribosomal leaders. Additionally, 23S methyl RNAs have rho-independent transcription terminators downstream, which could be a mechanism of regulation.
The 23S methyl RNA is found exclusively in the order of bacteria called Lactobacillales.


Margaret Anderson (indexer)

Margaret Dampier Anderson (née Whetham; 1900–1997) was a British biochemist and scientific indexer. She published four scientific articles in the 1920s before marrying in 1927 and began indexing books beginning in 1960.
Life.
Margaret Whetham was born on 21 April 1900, the daughter of William Cecil Dampier Whetham, a Cambridge-educated scientist and agricultural academic, and his wife Catherine Durning Holt, a daughter of Liverpool merchant Robert Durning Holt who had also pursued an education at Cambridge. One of her many great aunts (the “Potter sisters”) was social reformer Beatrice Webb. She had one brother and four sisters, including Edith Holt Whetham. Her family owned a small manor house in Devon, and also inherited a small estate in Hilfield, Dorset where they spent family vacations.
Margaret Whetham attended University College, Exeter and then earned her M.A. from Newnham College, Cambridge in 1926. While in graduate school, she worked with Marjory Stephenson including work on the washed suspension technique, which had originated with Louis Pasteur, for extracting enzymes from bacteria. Whetham co-authored four scientific papers with Stephenson and held the Old Students Jubilee Research Fellowship in 1926–27. She married Dr. Alan Bruce Anderson, a clinical pathologist, on 12 September 1927, and had five children and sixteen grandchildren including Tiffany Margaret Hall.
Work.
She abstracted scientific articles for several years before beginning to work as a freelance scientific indexer in 1960, creating indexes for 567 books over her career. Anderson joined the Society of Indexers two years later and served as its treasurer, membership secretary, member of the board of assessors, and vice-president over the next several decades. She was awarded the Wheatley Medal by the Society in 1975 for her index to "Copy-editing: The Cambridge Handbook". Eight years later, the Society presented her with its Carey Award for "outstanding services to indexing".


2-Phosphoglyceric acid

2-Phosphoglyceric acid (2PG), or 2-phosphoglycerate, is a glyceric acid which serves as the substrate in the ninth step of glycolysis. It is catalyzed by enolase into phosphoenolpyruvate (PEP), the penultimate step in the conversion of glucose to pyruvate.


Mycena kentingensis

Mycena kentingensis is a species of agaric fungus in the family Mycenaceae. Found in Taiwan and described as new to science in 2013, the fruit bodies of the fungus are bioluminescent. Closely related species include "M. stylobates" and "M. adscendens".


SDS-PAGE

SDS-PAGE (sodium dodecyl sulphate–polyacrylamide gel electrophoresis), is a discontinuous electrophoretic system developed by Ulrich K. Laemmli which is commonly used as a method to separate proteins with molecular masses between 5 and 250 kDa. The combined use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel allows to eliminate the influence of structure and charge, and proteins are separated solely on the basis of differences in their molecular weight.
Properties.
SDS-PAGE is an electrophoresis method that allows protein separation by mass. The medium (also referred to as ′matrix′) is a polyacrylamide-based discontinuous gel. The polyacrylamide-gel is typically sandwiched between two glass plates in a slab gel. Although tube gels (in glass cylinders) were used historically, they were rapidly made obsolete with the invention of the more convenient slab gels. In addition, SDS (sodium dodecyl sulfate) is used. About 1.4 grams of SDS bind to a gram of protein, corresponding to one SDS molecule per two amino acids. SDS acts as a surfactant, masking the proteins' intrinsic charge and conferring them very similar charge-to-mass ratios. The intrinsic charges of the proteins are negligible in comparison to the SDS loading, and the positive charges are also greatly reduced in the basic pH range of a separating gel. Upon application of a constant electric field, the protein migrate towards the anode, each with a different speed, depending on its mass. This simple procedure allows precise protein separation by mass.
SDS tends to form spherical micelles in aqueous solutions above a certain concentration called the critical micellar concentration (CMC). Above the critical micellar concentration of 7 to 10 millimolar in solutions, the SDS simultaneously occurs as single molecules (monomer) and as micelles, below the CMC SDS occurs only as monomers in aqueous solutions. At the critical micellar concentration, a micelle consists of about 62 SDS molecules. However, only SDS monomers bind to proteins via hydrophobic interactions, whereas the SDS micelles are anionic on the outside and do not adsorb any protein. SDS is amphipathic in nature, which allows it to unfold both polar and nonpolar sections of protein structure. In SDS concentrations above 0.1 millimolar, the unfolding of proteins begins, and above 1 mM, most proteins are denatured. Due to the strong denaturing effect of SDS and the subsequent dissociation of protein complexes, quaternary structures can generally not be determined with SDS. Exceptions are proteins that are stabilised by covalent cross-linking e.g. -S-S- linkages and the SDS-resistant protein complexes, which are stable even in the presence of SDS (the latter, however, only at room temperature). To denature the SDS-resistant complexes a high activation energy is required, which is achieved by heating. SDS resistance is based on a metastability of the protein fold. Although the native, fully folded, SDS-resistant protein does not have sufficient stability in the presence of SDS, the chemical equilibrium of denaturation at room temperature occurs slowly. Stable protein complexes are characterised not only by SDS resistance but also by stability against proteases and an increased biological half-life.
Alternatively, polyacrylamide gel electrophoresis can also be performed with the cationic surfactants CTAB in a CTAB-PAGE, or 16-BAC in a BAC-PAGE.
Procedure.
The SDS-PAGE method is composed of gel preparation, sample preparation, electrophoresis, protein staining or western blotting and analysis of the generated banding pattern.
Gel production.
When using different buffers in the gel (discontinuous gel electrophoresis), the gels are made up to one day prior to electrophoresis, so that the diffusion does not lead to a mixing of the buffers. The gel is produced by radical polymerisation in a mold consisting of two sealed glass plates with spacers between the glass plates. In a typical mini-gel setting, the spacers have a thickness of 0.75 mm or 1.5 mm, which determines the loading capacity of the gel. For pouring the gel solution, the plates are usually clamped in a stand which temporarily seals the otherwise open underside of the glass plates with the two spacers. For the gel solution, acrylamide is mixed as gel-former (usually 4% V/V in the stacking gel and 10-12 % in the separating gel), methylenebisacrylamide as a cross-linker, stacking or separating gel buffer, water and SDS. By adding the catalyst TEMED and the radical initiator ammonium persulfate (APS) the polymerisation is started. The solution is then poured between the glass plates without creating bubbles. Depending on the amount of catalyst and radical starter and depending on the temperature, the polymerisation lasts between a quarter of an hour and several hours. The lower gel (separating gel) is poured first and covered with a few drops of a barely water-soluble alcohol (usually buffer-saturated butanol or isopropanol), which eliminates bubbles from the meniscus and protects the gel solution of the radical scavenger oxygen. After the polymerisation of the separating gel, the alcohol is discarded and the residual alcohol is removed with filter paper. After addition of APS and TEMED to the stacking gel solution, it is poured on top of the solid separation gel. Afterwards, a suitable sample comb is inserted between the glass plates without creating bubbles. The sample comb is carefully pulled out after polymerisation, leaving pockets for the sample application. For later use of proteins for protein sequencing, the gels are often prepared the day before electrophoresis to reduce reactions of unpolymerised acrylamide with cysteines in proteins.
By using a gradient mixer, gradient gels with a gradient of acrylamide (usually from 4 to 12%) can be cast, which have a larger separation range of the molecular masses. Commercial gel systems (so-called "pre-cast gels") usually use the buffer substance Bis-tris methane with a pH value between 6.4 and 7.2 both in the stacking gel and in the separating gel. These gels are delivered cast and ready-to-use. Since they use only one buffer (continuous gel electrophoresis) and have a nearly neutral pH, they can be stored for several weeks. The more neutral pH slows the hydrolysis and thus the decomposition of the polyacrylamide. Furthermore, there are fewer acrylamide-modified cysteines in the proteins. Due to the constant pH in collecting and separating gel there is no stacking effect. Proteins in BisTris gels can not be stained with ruthenium complexes. This gel system has a comparatively large separation range, which can be varied by using MES or MOPS in the running buffer.
Sample preparation.
During sample preparation, the sample buffer, and thus SDS, is added in excess to the proteins, and the sample is then heated to 95 °C for five minutes, or alternatively 70 °C for ten minutes. Heating disrupts the secondary and tertiary structures of the protein by disrupting hydrogen bonds and stretching the molecules. Optionally, disulfide bridges can be cleaved by reduction. For this purpose, reducing thiols such as β-mercaptoethanol (β-ME, 5% by volume), dithiothreitol (DTT, 10 millimolar) or dithioerythritol (DTE, 10 millimolar) are added to the sample buffer. After cooling to room temperature, each sample is pipetted into its own well in the gel, which was previously immersed in electrophoresis buffer in the electrophoresis apparatus.
In addition to the samples, a molecular-weight size marker is usually loaded onto the gel. This consists of proteins of known sizes and thereby allows the estimation (with an error of ± 10%) of the sizes of the proteins in the actual samples, which migrate in parallel in different tracks of the gel. The size marker is often pipetted into the first or last pocket of a gel.
Electrophoresis.
For separation, the denatured samples are loaded onto a gel of polyacrylamide, which is placed in an electrophoresis buffer with suitable electrolytes. Thereafter, a voltage (usually around 100 V, 10-20 V per cm gel length) is applied, which causes a migration of negatively charged molecules through the gel in the direction of the positively charged anode. The gel acts like a sieve. Small proteins migrate relatively easily through the mesh of the gel, while larger proteins are more likely to be retained and thereby migrate more slowly through the gel, thereby allowing proteins to be separated by molecular size. The electrophoresis lasts between half an hour to several hours depending on the voltage and length of gel used.
The fastest-migrating proteins (with a molecular weight of less than 5 kDa) form the buffer front together with the anionic components of the electrophoresis buffer, which also migrate through the gel. The area of the buffer front is made visible by adding the comparatively small, anionic dye bromophenol blue to the sample buffer. Due to the relatively small molecule size of bromophenol blue, it migrates faster than proteins. By optical control of the migrating colored band, the electrophoresis can be stopped before the dye and also the samples have completely migrated through the gel and leave it.
The most commonly used method is the discontinuous SDS-PAGE. In this method, the proteins migrate first into a collecting gel with neutral pH, in which they are concentrated and then they migrate into a separating gel with basic pH, in which the actual separation takes place. Stacking and separating gels differ by different pore size (4-6 % T and 10-20 % T), ionic strength and pH values (pH 6.8 or pH 8.8). The electrolyte most frequently used is an SDS-containing Tris-glycine-chloride buffer system. At neutral pH, glycine predominantly forms the zwitterionic form, at high pH the glycines lose positive charges and become predominantly anionic. In the collection gel, the smaller, negatively charged chloride ions migrate in front of the proteins (as leading ions) and the slightly larger, negatively and partially positively charged glycinate ions migrate behind the proteins (as initial trailing ions), whereas in the comparatively basic separating gel both ions migrate in front of the proteins. The pH gradient between the stacking and separation gel buffers leads to a stacking effect at the border of the stacking gel to the separation gel, since the glycinate partially loses its slowing positive charges as the pH increases and then, as the former trailing ion, overtakes the proteins and becomes a leading ion, which causes the bands of the different proteins (visible after a staining) to become narrower and sharper - the stacking effect. For the separation of smaller proteins and peptides, the TRIS-Tricine buffer system of Schägger and von Jagow is used due to the higher spread of the proteins in the range of 0.5 to 50 kDa.
Gel staining.
At the end of the electrophoretic separation, all proteins are sorted by size and can then be analyzed by other methods, e. g. protein staining such as Coomassie staining (most common and easy to use), silver staining (highest sensitivity), stains all staining, Amido black 10B staining, Fast green FCF staining, fluorescent stains such as epicocconone stain and SYPRO orange stain, and immunological detection such as the Western Blot. The fluorescent dyes have a comparatively higher linearity between protein quantity and color intensity of about three orders of magnitude above the detection limit, i. e. the amount of protein can be estimated by color intensity. When using the fluorescent protein dye trichloroethanol, a subsequent protein staining is omitted if it was added to the gel solution and the gel was irradiated with UV light after electrophoresis.
In Coomassie Staining, Gel is Fixed in a 50% ethanol 10% glacial acetic acid solution for 1 hr. Then the solution is changed for fresh one and after 1 to 12 hrs Gel is changed to a Staining solution (50% methanol, 10% Glacial acetic Acid, 0.1% Coomassie Brilliant Blue) followed by destaining changing several times a destaining solution of 40% methanol, 10% glacial acetic acid.
Analysis.
Protein staining in the gel creates a documentable banding pattern of the various proteins. Glycoproteins have differential levels of glycosylations and adsorb SDS more unevenly at the glycosylations, resulting in broader and blurred bands. Membrane proteins, because of their transmembrane domain, are often composed of the more hydrophobic amino acids, have lower solubility in aqueous solutions, tend to bind lipids, and tend to precipitate in aqueous solutions due to hydrophobic effects when sufficient amounts of detergent are not present. This precipitation manifests itself for membrane proteins in a SDS-PAGE in "tailing" above the band of the transmembrane protein. In this case, more SDS can be used (by using more or more concentrated sample buffer) and the amount of protein in the sample application can be reduced. An overloading of the gel with a soluble protein creates a semicircular band of this protein (e. g. in the marker lane of the image at 66 kDa), allowing other proteins with similar molecular weights to be covered. A low contrast (as in the marker lane of the image) between bands within a lane indicates either the presence of many proteins (low purity) or, if using purified proteins and a low contrast occurs only below one band, it indicates a proteolytic degradation of the protein, which first causes degradation bands, and after further degradation produces a homogeneous color ("smear") below a band. The documentation of the banding pattern is usually done by photographing or scanning. For a subsequent recovery of the molecules in individual bands, a gel extraction can be performed.
Archiving.
After protein staining and documentation of the banding pattern, the polyacrylamide gel can be dried for archival storage. Proteins can be extracted from it at a later date. The gel is either placed in a drying frame (with or without the use of heat) or in a vacuum dryer. The drying frame consists of two parts, one of which serves as a base for a wet cellophane film to which the gel and a one percent glycerol solution are added. Then a second wet cellophane film is applied bubble-free, the second frame part is put on top and the frame is sealed with clips. The removal of the air bubbles avoids a fragmentation of the gel during drying. The water evaporates through the cellophane film. In contrast to the drying frame, a vacuum dryer generates a vacuum and heats the gel to about 50 °C.
Molecular mass determination.
For a more accurate determination of the molecular weight, the relative migration distances of the individual protein bands are measured in the separating gel. The measurements are usually performed in triplicate for increased accuracy. The relative mobility (called Rf value or Rm value) is the quotient of the distance of the band of the protein and the distance of the buffer front. The distances of the bands and the buffer front are each measured from the beginning of the separation gel. The distance of the buffer front roughly corresponds to the distance of the bromophenol blue contained in the sample buffer. The relative distances of the proteins of the size marker are plotted semi-logarithmically against their known molecular weights. By comparison with the linear part of the generated graph or by a regression analysis, the molecular weight of an unknown protein can be determined by its relative mobility. Bands of proteins with glycosylations can be blurred. Proteins with many basic amino acids (e. g. histones) can lead to an overestimation of the molecular weight or even not migrate into the gel at all, because they move slower in the electrophoresis due to the positive charges or even to the opposite direction. Accordingly, many acidic amino acids can lead to accelerated migration of a protein and an underestimation of its molecular mass.
Applications.
The SDS-PAGE in combination with a protein stain is widely used in biochemistry for the quick and exact separation and subsequent analysis of proteins. It has comparatively low instrument and reagent costs and is an easy-to-use method. Because of its low scalability, it is mostly used for analytical purposes and less for preparative purposes, especially when larger amounts of a protein are to be isolated.
Additionally, SDS-PAGE is used in combination with the western blot for the determination of the presence of a specific protein in a mixture of proteins - or for the analysis of post-translational modifications. Post-translational modifications of proteins can lead to a different relative mobility (i.e. a "band shift") or to a change in the binding of a detection antibody used in the western blot (i.e. a band disappears or appears).
In mass spectrometry of proteins, SDS-PAGE is a widely used method for sample preparation prior to spectrometry, mostly using in-gel digestion. In regards to determining the molecular mass of a protein, the SDS-PAGE is a bit more exact than an analytical ultracentrifugation, but less exact than a mass spectrometry or - ignoring post-translational modifications - a calculation of the protein molecular mass from the DNA sequence.
In medical diagnostics, SDS-PAGE is used as part of the HIV test and to evaluate proteinuria. In the HIV test, HIV proteins are separated by SDS-PAGE and subsequently detected by Western Blot with HIV-specific antibodies of the patient, if they are present in his blood serum. SDS-PAGE for proteinuria evaluates the levels of various serum proteins in the urine, e.g. Albumin, Alpha-2-macroglobulin and IgG.
Variants.
SDS-PAGE is the most widely used method for gel electrophoretic separation of proteins. Two-dimensional gel electrophoresis sequentially combines isoelectric focusing or BAC-PAGE with a SDS-PAGE. Native PAGE is used if native protein folding is to be maintained. For separation of membrane proteins, BAC-PAGE or CTAB-PAGE may be used as an alternative to SDS-PAGE. For electrophoretic separation of larger protein complexes, agarose gel electrophoresis can be used, e.g. the SDD-AGE. Some enzymes can be detected via their enzyme activity by zymography.
Alternatives.
While being one of the more precise and low-cost protein separation and analysis methods, the SDS-PAGE denatures proteins. Where non-denaturing conditions are necessary, proteins are separated by a native PAGE or different chromatographic methods with subsequent photometric quantification, for example affinity chromatography (or even tandem affinity purification), size exclusion chromatography, ion exchange chromatography. Proteins can also be separated by size in a tangential flow filtration or an ultrafiltration. Single proteins can be isolated from a mixture by affinity chromatography or by a pull-down assay. Some historically early and cost effective but crude separation methods usually based upon a series of extractions and precipitations using kosmotropic molecules, for example the ammonium sulfate precipitation and the polyethyleneglycol precipitation.
History.
In 1948, Arne Tiselius was awarded the Nobel Prize in Chemistry for the discovery of the principle of electrophoresis as the migration of charged and dissolved atoms or molecules in an electric field. The use of a solid matrix (initially paper discs) in a zone electrophoresis improved the separation. The discontinuous electrophoresis of 1964 by L. Ornstein and B. J. Davis made it possible to improve the separation by the stacking effect. The use of cross-linked polyacrylamide hydrogels, in contrast to the previously used paper discs or starch gels, provided a higher stability of the gel and no microbial decomposition. The denaturing effect of SDS in continuous polyacrylamide gels and the consequent improvement in resolution was first described in 1965 by David F. Summers in the working group of James E. Darnell to separate poliovirus proteins. The current variant of the SDS-PAGE was described in 1970 by Ulrich K. Laemmli and initially used to characterise the proteins in the head of bacteriophage T4. This Laemmli paper is widely cited for the invention of modern SDS-PAGE, but the technique was actually invented by Jake Maizel, who was doing a sabbatical in the MRC laboratory when Laemmli joined the lab as a postdoctoral fellow. Maizel shared his prior technology with Laemmli and together they made further improvements. Laemmli and Maizel had planned to follow up with a Methods paper but this never materialized. Maizel recounts the history of development of SDS-PAGE in brief commentary.


Trends in Cell Biology


Phosphoadenylyl-sulfate reductase (thioredoxin)

In enzymology, a phosphoadenylyl-sulfate reductase (thioredoxin) () is an enzyme that catalyzes the chemical reaction
The 3 substrates of this enzyme are adenosine 3',5'-bisphosphate, sulfite, and thioredoxin disulfide, whereas its two products are 3'-phosphoadenylyl sulfate and thioredoxin.
This enzyme belongs to the family of oxidoreductases, specifically those acting on a sulfur group of donors with a disulfide as acceptor. The systematic name of this enzyme class is adenosine 3',5'-bisphosphate,sulfite:thioredoxin-disulfide oxidoreductase (3'-phosphoadenosine-5'-phosphosulfate-forming). Other names in common use include PAPS reductase, thioredoxin-dependent, PAPS reductase, thioredoxin:adenosine 3'-phosphate 5'-phosphosulfate reductase, 3'-phosphoadenylylsulfate reductase, thioredoxin:3'-phospho-adenylylsulfate reductase, phosphoadenosine-phosphosulfate reductase, adenosine 3',5'-bisphosphate,sulfite:oxidized-thioredoxin, and oxidoreductase (3'-phosphoadenosine-5'-phosphosulfate-forming). This enzyme participates in sulfur metabolism.
Structural studies.
As of late 2007, 3 structures have been solved for this class of enzymes, with PDB accession codes , , and .


CsrB/RsmB RNA family

The CsrB RNA is a non-coding RNA that binds to approximately 9 to 10 dimers of the CsrA protein. The CsrB RNAs contain a conserved motif CAGGXXG that is found in up to 18 copies and has been suggested to bind CsrA. The Csr regulatory system has a strong negative regulatory effect on glycogen biosynthesis, glyconeogenesis and glycogen catabolism and a positive regulatory effect on glycolysis. In other bacteria such as "Erwinia carotovora" the RsmA protein has been shown to regulate the production of virulence determinants, such extracellular enzymes. RsmA binds to RsmB regulatory RNA which is also a member of this family.
RsmB RNA was shown to be upregulated by GacS/A system, and increase downstream T3SS gene expression. FlhDC, the master regulator of flagellar genes, also activates rsmB RNA production. A regulatory network have been revealed connecting rsmB, FlhDC and T3SS.
It has been shown to play role in the biocontrol activity of "Rahnella aquatilis" HX2 (a biocontrol agent producing antibacterial substance).