Flavin-containing monooxygenase

The flavin-containing monooxygenase (FMO) protein family specializes in the oxidation of xeno-substrates in order to facilitate the excretion of these compounds from living organisms. These enzymes can oxidize a wide array of heteroatoms, particularly soft nucleophiles, such as amines, sulfides, and phosphites. This reaction requires an oxygen, an NADPH cofactor, and an FAD prosthetic group. FMOs share several structural features, such as a NADPH binding domain, FAD binding domain, and a conserved arginine residue present in the active site. Recently, FMO enzymes have received a great deal of attention from the pharmaceutical industry both as a drug target for various diseases and as a means to metabolize pro-drug compounds into active pharmaceuticals. These monooxygenases are often misclassified because they share activity profiles similar to those of cytochrome P450 (CYP450), which is the major contributor to oxidative xenobiotic metabolism. However, a key difference between the two enzymes lies in how they proceed to oxidize their respective substrates; CYP enzymes make use of an oxygenated heme prosthetic group, while the FMO family utilizes FAD to oxidize its substrates.
History.
Prior to the 1960s, the oxidation of xenotoxic materials was thought to be completely accomplished by CYP450. However, in the early 1970s, Dr. Daniel Ziegler from the University of Texas at Austin discovered a hepatic flavoprotein isolated from pig liver that was found to oxidize a vast array of various amines to their corresponding nitro state. This flavoprotein named "Ziegler's enzyme" exhibited unusual chemical and spectrometric properties. Upon further spectroscopic characterization and investigation of the substrate pool of this enzyme, Dr. Ziegler discovered that this enzyme solely bound FAD molecule that could form a C4a-hydroxyperoxyflavin intermediate, and that this enzyme could oxidize a wide variety of substrates with no common structural features, including phosphines, sulfides, selenium compounds, amongst others. Once this was noticed, Dr. Ziegler's enzyme was reclassified as a broadband flavin monooxygenase.
In 1984, the first evidence for multiple forms of FMOs was elucidated by two different laboratories when two distinct FMOs were isolated from rabbit lungs. Since then, over 150 different FMO enzymes have been successfully isolated from a wide variety of organisms. Up until 2002, only 5 FMO enzymes were successfully isolated from mammals. However, a group of researchers found a sixth FMO gene located on human chromosome 1. In addition to the sixth FMO discovered as of 2002, the laboratories of Dr. Ian Philips and Elizabeth Sheppard discovered a second gene cluster in humans that consists of 5 additional pseudogenes for FMO on human chromosome 1.
Evolution of FMO gene family.
The FMO family of genes is conserved across all phyla that have been studied so far, therefore some form of the FMO gene family can be found in all studied eukaryotes. FMO genes are characterized by specific structural and functional constraints, which led to the evolution of different types of FMO's in order to perform a variety of functions. Divergence between the functional types of FMO's (FMO 1–5) occurred before the amphibians and mammals diverged into separate classes. FMO5 found in vertebrates appears to be evolutionarily older than other types of FMO's, making FMO5 the first functionally distinct member of the FMO family. Phylogenetic studies suggest that FMO1 and FMO3 are the most recent FMO's to evolve into enzymes with distinct functions. Although FMO5 was the first distinct FMO, it is not clear what function it serves since it does not oxygenate the typical FMO substrates involved in first-pass metabolism.
Analyses of FMO genes across several species have shown extensive silent DNA mutations, which indicate that the current FMO gene family exists because of selective pressure at the protein level rather than the nucleotide level. FMO's found in invertebrates are found to have originated polyphyletically; meaning that a phenotypically similar gene evolved in invertebrates which was not inherited from a common ancestor.
Classification and characterization.
FMOs are one subfamily of class B external flavoprotein monooxygenases (EC 1.14.13), which belong to the family of monooxygenase oxidoreductases, along with the other subfamilies Baeyer-Villiger monooxygenases and microbial N-hydroxylating monooxygenases. FMO's are found in fungi, yeast, plants, mammals, and bacteria.
Mammals.
Developmental and tissue specific expression has been studied in several mammalian species, including humans, mice, rats, and rabbits. However, because FMO expression is unique to each animal species, it is difficult to make conclusions about human FMO regulation and activity based on other mammalian studies. It is likely that species-specific expression of FMO's contributes to differences in susceptibility to toxins and xenobiotics as well as the efficiency with excreting among different mammals.
Six functional forms of human FMO genes have been reported. However, FMO6 is considered to be a pseudogene. FMOs 1–5 share between 50–58% amino acid identity across the different species. Recently, five more human FMO genes were discovered, although they fall in the category of pseudogenes.
Yeast.
Unlike mammals, yeast ("Saccharomyces cerevisiae") do not have several isoforms of FMO, but instead only have one called yFMO. This enzyme does not accept xenobiotic compounds. Instead, yFMO helps to fold proteins that contain disulfide bonds by catalyzing O2 and NADPH-dependent oxidations of biological thiols, just like mammalian FMO's. An example is the oxidation of glutathione to glutathione disulfide, both of which form a redox buffering system in the cell between the endoplasmic reticulum and the cytoplasm. yFMO is localized in the cytoplasm in order to maintain the optimum redox buffer ratio necessary for proteins containing disulfide bonds to fold properly. This non-xenobiotic role of yFMO may represent the original role of the FMO's before the rise of the modern FMO family of enzymes found in mammals.
Plants.
Plant FMO's play a role in defending against pathogens and catalyze specific steps in the biosynthesis of auxin, a plant hormone. Plant FMO's also play a role in the metabolism of glucosinolates. These non-xenobiotic roles of plant FMO's suggest that other FMO functions could be identified in non-plant organisms.
Structure.
Crystal structures have been determined for yeast ("Schizosaccharomyces pombe") FMO (PDB: 1VQW) and bacterial ("Methylophaga aminisulfidivorans") FMO (PDB: 2XVH). The crystal structures are similar to each other and they share 27% sequence identity. These enzymes share 22% and 31% sequence identity with human FMOs, respectively.
FMOs have a tightly bound FAD prosthetic group and a binding NADPH cofactor. Both dinucleotide binding motifs form Rossmann folds. The yeast FMO and bacterial FMO are dimers, with each monomer consisting of two structural domains: the smaller NADPH binding domain and the larger FAD-binding domain. The two domains are connected by a double linker. A channel between the two domains leads to the active site where NADPH binds both domains and occupies a cleft that blocks access to the flavin group of FAD, which is bound to the large domain along the channel together with a water molecule. The nicotinamide group of NADPH interacts with the flavin group of FAD, and the NADPH binding site overlaps with the substrate binding site on the flavin group.
FMOs contain several sequence motifs that are conserved across all domains:
The FMO identifying motif interacts with the flavin of FAD. The F/LATGY motif is a sequence motif common in "N"-hydroxylating enzymes. The arginine residue interacts with the phosphate group of NADPH.
Function.
The general function of these enzymes is to metabolise xenobiotics. Hence, they are considered to be xenobiotic detoxication catalysts. These proteins catalyze the oxygenation of multiple heteroatom-containing compounds that are present in our diet, such as amine-, sulfide-, phosphorus-, and other nucleophilic heteroatom-containing compounds. FMOs have been implicated in the metabolism of a number of pharmaceuticals, pesticides and toxicants, by converting the lipophilic xenobiotics into polar, oxygenated, and readily excreted metabolites.
Substrate diversity.
FMO substrates are structurally diverse compounds. However, they all share similar characteristics:
Zwitterions, anions and dications are considered to be unfavorable substrates. There are several drugs reported to be typical substrates for FMOs.
The majority of drugs function as alternate substrate competitive inhibitors to FMOs (i.e. good nucleophiles that compete with the drug for FMO oxygenation), since they are not likely to serve as FMO substrates. Only a few true FMO competitive inhibitors have been reported. Those include indole-3-carbinol and "N","N"-dimethylamino stilbene carboxylates. A well-known FMO inhibitor is methimazole (MMI).
Mechanism.
The FMO catalytic cycle proceeds as follows:
Cellular expression in humans.
Expression of each type of FMO relies on several factors including, cofactor supply, physiological & environmental factors, as well as diet. Because of these factors, each type of FMO is expressed differently depending on the species and tissue. In humans, expression of FMO's is mainly concentrated to the human liver, lungs, and kidneys, where most of the metabolism of xenobiotics occur. However, FMO's can also be found in the human brain and small intestine. While FMO1-5 can be found in the brain, liver, kidneys, lungs, and small intestine, the distribution of each type of FMO differs depending on the tissue and the developmental stage of the person.
Expression in adult tissues.
In an adult, FMO1 is predominately expressed in the kidneys and to a lesser extent in the lungs and small intestine. FMO2 is the most abundant of the FMO's and is mostly expressed in the lungs and kidneys, with lower expression in the liver and small intestine. FMO3 is highly concentrated in the liver, but is also expressed in the lungs. FMO4 is expressed mostly in the liver and kidneys. FMO5 is highly expressed in the liver, but also has substantial expression in the lungs and small intestine. Though FMO2 is the most expressed FMO in the brain, it only constitutes about 1% of that found in the lungs, making FMO expression in the brain fairly low.
Expression in fetal Tissues.
The distribution of FMO's in various types of tissues changes as a person continues to develop, making the fetal distribution of FMO's quite different than adult distribution of FMO's. While the adult liver is dominated by the expression of FMO3 and FMO5, the fetal liver is dominated by the expression of FMO1 and FMO5. Another difference is in the brain, where adults mostly express FMO2 and fetuses mostly express FMO1.
Clinical significance.
Drug development.
Drug metabolism is one of the most important factors to consider when developing new drugs for therapeutic applications. The degradation rate of these new drugs in an organism's system determines the duration and intensity of their pharmacological action. During the past few years, FMOs have gained a lot of attention in drug development since these enzymes are not readily induced or inhibited by the chemicals or drugs surrounding their environment. CYPs are the primary enzymes involved in drug metabolism. However, recent efforts have been directed towards the development of drug candidates that incorporate functional groups that can be metabolized by FMOs. By doing this, the number of potential adverse drug-drug interactions is minimized and the reliance on CYP450 metabolism is decreased. Several approaches have been made to screen potential drug interactions. One of them includes human FMO3 (hFMO3), which is described as the most vital FMO regarding drug interactions. In order to successfully screen hFMO3 in a high throughput fashion hFMO3 was successfully fixed to graphene oxide chips in order to measure the change in electrical potential generated as a result of the drug being oxidized when it interacts with the enzyme.
Hypertension.
There is evidence that FMOs are associated to the regulation of blood pressure. FMO3 is involved in the formation of TMA N-oxides (TMAO). Some studies indicate that hypertension can develop when there are no organic osmolytes (i.e. TMAO) that can counteract an increase in osmotic pressure and peripheral resistance. Individuals with deficient FMO3 activity have a higher prevalence of hypertension and other cardiovascular diseases, since there is a decrease in formation of TMA N-oxides to counterbalance the effects of a higher osmotic pressure and peripheral resistance.
Fish odor syndrome.
The trimethylaminuria disorder, also known as fish odor syndrome, causes abnormal FMO3-mediated metabolism or a deficiency of this enzyme in an individual. A person with this disorder has a low capacity to oxidize the trimethylamine (TMA) that comes from their diet to its odourless metabolite TMAO. When this happens, large amounts of TMA are excreted through the individual's urine, sweat, and breath, with a strong fish-like odor. As of today, there is no known cure or treatment for this disorder. However, doctors recommend patients to avoid foods containing choline, carnitine, nitrogen, sulfur and lecithin.
Other diseases.
FMOs have also been associated with other diseases, such as cancer and diabetes. Yet, additional studies are imperative to elucidate what is the relationship between FMO function and these diseases, as well as to define these enzymes’ clinical relevance.


A-86929

A-86929 is a synthetic compound that acts as a selective dopamine receptor D1 agonist. It was developed as a possible treatment for Parkinson's disease, as well as for other applications such as treatment of cocaine addiction, but while it had reasonable efficacy in humans it also caused dyskinesias and has not been continued. It has mainly been used as its diacetate ester prodrug adrogolide (ABT-431), which has better bioavailability.


ATPC RNA motif

The ATPC RNA motif is a conserved RNA structure found in certain cyanobacteria. It is apparently ubiquitous in "Prochlorococcus marinus", and is present in many species in the genus "Synechococcus". The RNA is always found within an operon encoding subunits of ATP synthase, and it is always located downstream of the gene encoding the A subunit of ATP synthase, and upstream of the C subunit gene. This location is consistent with a cis-regulatory element, but also with a non-coding RNA that is transcribed with the ATP synthase genes.
Simple RNA structures called stem-loops have been reported in the ATP synthase operons of various cyanobacteria, but not structures such as the 3-stem junction that is the main feature of the ATPC RNA motif.


Sequence assembly

In bioinformatics, sequence assembly refers to aligning and merging fragments from a longer DNA sequence in order to reconstruct the original sequence. This is needed as DNA sequencing technology cannot read whole genomes in one go, but rather reads small pieces of between 20 and 30,000 bases, depending on the technology used. Typically the short fragments, called reads, result from shotgun sequencing genomic DNA, or gene transcript (ESTs).
The problem of sequence assembly can be compared to taking many copies of a book, passing each of them through a shredder with a different cutter, and piecing the text of the book back together just by looking at the shredded pieces. Besides the obvious difficulty of this task, there are some extra practical issues: the original may have many repeated paragraphs, and some shreds may be modified during shredding to have typos. Excerpts from another book may also be added in, and some shreds may be completely unrecognizable.
Genome assemblers.
The first sequence assemblers began to appear in the late 1980s and early 1990s as variants of simpler sequence alignment programs to piece together vast quantities of fragments generated by automated sequencing instruments called DNA sequencers. As the sequenced organisms grew in size and complexity (from small viruses over plasmids to bacteria and finally eukaryotes), the assembly programs used in these genome projects needed increasingly sophisticated strategies to handle:
Faced with the challenge of assembling the first larger eukaryotic genomes—the fruit fly "Drosophila melanogaster" in 2000 and the human genome just a year later,—scientists developed assemblers like Celera Assembler and Arachne able to handle genomes of 130 million (e.g., the fruit fly "D. melanogaster") to 3 billion (e.g., the human genome) base pairs. Subsequent to these efforts, several other groups, mostly at the major genome sequencing centers, built large-scale assemblers, and an open source effort known as AMOS was launched to bring together all the innovations in genome assembly technology under the open source framework.
EST assemblers.
Expressed sequence tag or EST assembly was an early strategy, dating from the mid-1990s to the mid-2000s, to assemble individual genes rather than whole genomes. The problem differs from genome assembly in several ways. The input sequences for EST assembly are fragments of the transcribed mRNA of a cell and represent only a subset of the whole genome. A number of algorithmical problems differ between genome and EST assembly. For instance, genomes often have large amounts of repetitive sequences, concentrated in the intergenic regions. Transcribed genes contain many fewer repeats, making assembly somewhat easier. On the other hand, some genes are expressed (transcribed) in very high numbers (e.g., housekeeping genes), which means that unlike whole-genome shotgun sequencing, the reads are not uniformly sampled across the genome.
EST assembly is made much more complicated by features like (cis-) alternative splicing, trans-splicing, single-nucleotide polymorphism, and post-transcriptional modification. Beginning in 2008 when RNA-Seq was invented, EST sequencing was replaced by this far more efficient technology, described under de novo transcriptome assembly.
De-novo vs. mapping assembly.
In sequence assembly, two different types can be distinguished:
In terms of complexity and time requirements, de-novo assemblies are orders of magnitude slower and more memory intensive than mapping assemblies. This is mostly due to the fact that the assembly algorithm needs to compare every read with every other read (an operation that has a naive time complexity of O(n2). Referring to the comparison drawn to shredded books in the introduction: while for mapping assemblies one would have a very similar book as a template (perhaps with the names of the main characters and a few locations changed), de-novo assemblies present a more daunting challenge in that one would not know beforehand whether this would become a science book, a novel, a catalogue, or even several books. Also, every shred would be compared with every other shred.
Handling repeats in de-novo assembly requires the construction of a graph representing neighboring repeats. Such information can be derived from reading a long fragment covering the repeats in full or only its two ends. On the other hand, in a mapping assembly, parts with multiple or no matches are usually left for another assembling technique to look into.
Influence of technological changes.
The complexity of sequence assembly is driven by two major factors: the number of fragments and their lengths. While more and longer fragments allow better identification of sequence overlaps, they also pose problems as the underlying algorithms show quadratic or even exponential complexity behaviour to both number of fragments and their length. And while shorter sequences are faster to align, they also complicate the layout phase of an assembly as shorter reads are more difficult to use with repeats or near identical repeats.
In the earliest days of DNA sequencing, scientists could only gain a few sequences of short length (some dozen bases) after weeks of work in laboratories. Hence, these sequences could be aligned in a few minutes by hand.
In 1975, the "dideoxy termination" method (AKA "Sanger sequencing") was invented and until shortly after 2000, the technology was improved up to a point where fully automated machines could churn out sequences in a highly parallelised mode 24 hours a day. Large genome centers around the world housed complete farms of these sequencing machines, which in turn led to the necessity of assemblers to be optimised for sequences from whole-genome shotgun sequencing projects where the reads
With the Sanger technology, bacterial projects with 20,000 to 200,000 reads could easily be assembled on one computer. Larger projects, like the human genome with approximately 35 million reads, needed large computing farms and distributed computing.
By 2004 / 2005, pyrosequencing had been brought to commercial viability by 454 Life Sciences. This new sequencing method generated reads much shorter than those of Sanger sequencing: initially about 100 bases, now 400-500 bases. Its much higher throughput and lower cost (compared to Sanger sequencing) pushed the adoption of this technology by genome centers, which in turn pushed development of sequence assemblers that could efficiently handle the read sets. The sheer amount of data coupled with technology-specific error patterns in the reads delayed development of assemblers; at the beginning in 2004 only the Newbler assembler from 454 was available. Released in mid-2007, the hybrid version of the MIRA assembler by Chevreux et al. was the first freely available assembler that could assemble 454 reads as well as mixtures of 454 reads and Sanger reads. Assembling sequences from different sequencing technologies was subsequently coined "hybrid assembly".
From 2006, the Illumina (previously Solexa) technology has been available and can generate about 100 million reads per run on a single sequencing machine. Compare this to the 35 million reads of the human genome project which needed several years to be produced on hundreds of sequencing machines. Illumina was initially limited to a length of only 36 bases, making it less suitable for de novo assembly (such as de novo transcriptome assembly), but newer iterations of the technology achieve read lengths above 100 bases from both ends of a 3-400bp clone. Announced at the end of 2007, the SHARCGS assembler by Dohm et al. was the first published assembler that was used for an assembly with Solexa reads. It was quickly followed by a number of others.
Later, new technologies like SOLiD from Applied Biosystems, Ion Torrent and SMRT were released and new technologies (e.g. Nanopore sequencing) continue to emerge. Despite the higher error rates of these technologies they are important for assembly because their longer read length helps to address the repeat problem. It is impossible to assemble through a perfect repeat that is longer than the maximum read length; however, as reads become longer the chance of a perfect repeat that large becomes small. This gives longer sequencing reads an advantage in assembling repeats even if they have low accuracy (~85%).
Greedy algorithm.
Given a set of sequence fragments, the object is to find a longer sequence that contains all the fragments.
The result need not be an optimal solution to the problem.
Programs.
For a lists of "de-novo" assemblers, see De novo sequence assemblers. For a list of mapping aligners, see List of sequence alignment software § Short-read sequence alignment.


Martin F. Jarrold

Martin F. Jarrold is a physical and analytical chemist known for contributions to ion-mobility spectrometry, heat capacity measurements of metal clusters, and charge detection mass spectrometry. Martin is the Robert & Marjorie Mann Chair in the Department of Chemistry at Indiana University.
Background.
Martin F. Jarrold grew up in England. He attended the University of Warwick in Coventry, England and obtained both a Bachelor of Science (1977) and PhD (1980). Jarrold came to the United States as a NATO Postdoctoral Fellow at the University of California in Santa Barbara. Joining the Physics Research Division of AT&T Bell Laboratories in Murray Hill, New Jersey after UCSB. There, he focused on determining the physical and chemical properties of semiconductor clusters. In 1992, he became Professor of Chemistry at Northwestern University, and was later named Dow Chemical Company Research Professor in 2000. While at Northwestern, he worked on advanced mass spectrometry-based-methods to study peptides and proteins. In the summer of 2002, he moved to Indiana University as Professor and a Robert & Marjorie Mann Chair in the Department of Chemistry.
Research.
Jarrold's research focuses include use of ion-mobility spectrometry to analyze proteins, peptides, clusters, and other biomolecules. Through ion mobility spectrometry, his group was able to see deviations from the native state and different conformations (different protein folding modes). Some of Jarrold's more recent research has investigated the formation of viral capsids by analyzing the intermediates in their formation with charge detection mass spectrometry (CDMS). Jarrold has been involved in the publication of over 250 articles.
Awards.
On June 8, 2018, Jarrold received the John B. Fenn Award at the American Society for Mass Spectrometry Conference for his work in ion mobility spectrometry. This award is given to those who have made distinguished contributions to the mass spectrometry field.


Molecular Biotechnology

Molecular Biotechnology is a peer-reviewed scientific journal published by Springer Science+Business Media. It publishes original research papers and review articles on the application of molecular biology to biotechnology. It was established in 1994 with John M. Walker as founding editor-in-chief. He was succeeded in 2011 by Giancarlo Basile.
Abstracting and indexing.
The journal is abstracted and indexed in:
According to the "Journal Citation Reports", the journal has a 2013 impact factor of 2.275.


Everton Genealogy Collection

The Everton Genealogy Collection was a noncirculating collection of genealogy and local history items that was part of the Logan Library in Logan, Utah. It included approximately 80,000 books, serials, pamphlets, compact discs, microfilms, atlases, maps, and databases. A large portion consisted of unpublished manuscripts that were rare or unique to the collection. The materials in the collection related to every state in the United States and the District of Columbia. While the focus was on the United States, there were resources for other nations as well.
History.
The Everton Genealogy Collection was started in 1947 by Walter Everton when he established the Everton Publishing Company. He and his successors in the company gathered items to be included in this collection. Some were donated, but most were added when they were sent to be reviewed in their magazine, "The Genealogical Helper". Over the course of nearly sixty years, the collection gradually grew to its present size.
In 2001, the publishing company was purchased by the Family History Network, Inc. and in 2004, then owner Bill Schjelderup, donated the collection to Logan City. In October 2006 the collection opened to the public in the former Logan Justice building. In early 2008, the collection was closed to the public while city departments shuffled buildings.
Current Status.
In 2010, a condensed collection opened to the public within the Logan Library in the space formerly occupied by the Logan Municipal Council. It is no longer identified as the Everton Collection but continues to be a large portion of the items housed in the library's Virginia Hansen Special Collections Room.


Pamatolol

Pamatolol is a beta adrenergic receptor antagonist.


Viscumamide

Viscumamide is a cyclic peptide isolated from endophytic fungi of mangrove.


TB9Cs3H2 snoRNA

TB9Cs3H2 is a member of the H/ACA-like class of non-coding RNA (ncRNA) molecule that guide the sites of modification of uridines to pseudouridines of substrate RNAs. It is known as a small nucleolar RNA (snoRNA) thus named because of its cellular localization in the nucleolus of the eukaryotic cell. TB9Cs3H2 is predicted to guide the pseudouridylation of LSU5 ribosomal RNA (rRNA) at residue Ψ1103.


Semaglutide

Semaglutide, sold under the brand names Ozempic and Rybelsus, is an anti-diabetic medication used for the treatment of type 2 diabetes.
Semaglutide acts like human glucagon-like peptide-1 (GLP-1) such that it increases insulin secretion, thereby increasing sugar metabolism. It is distributed as a metered subcutaneous injection in a prefilled pen or as an oral form. One of its advantages over other antidiabetic drugs is that it has a long duration of action, thus, only once-a-week injection is sufficient.
An injection version (Ozempic) was approved for medical use in the United States in December 2017, and in the European Union, Canada, and Japan in 2018. A version which is taken by mouth (Rybelsus) was approved for medical use in the United States in September 2019, and in the European Union in April 2020. It is the first glucagon-like peptide receptor protein treatment approved for use in the United States that does not need to be injected. It was developed by Novo Nordisk.
Side effects including nausea, vomiting, diarrhea, abdominal pain, and constipation may occur.
Medical uses.
Semaglutide is prepared for subcutaneous injection and is available in prefilled pen. It is recommended for once-weekly injection.
Adverse effects.
Side effects including nausea, vomiting, diarrhea, abdominal pain, and constipation may occur. In people with heart problems, it can cause damage to the back of the eye (retinopathy).
Side effects include medullary thyroid cancer, kidney problems, diabetic retinopathy, allergic reactions, low blood sugar, and pancreatitis.
Mechanism of action.
Semaglutide is a glucagon-like peptide-1 receptor agonist. It increases the production of insulin, a hormone that lowers the blood sugar level. It also appears to enhance growth of β cells in the pancreas, which are the sites of insulin production. On the other hand it inhibits glucagon, which is a hormone that increases blood sugar. It additionally reduces food intake by lowering appetite and slows down digestion in the stomach. In this way it works in body fat reduction.
Structure.
Semaglutide is chemically similar to human glucagon-like peptide-1 (GLP-1), with 94% similarity. The only differences are two amino acid substitutions at positions 8 and 34, where alanine and lysine are replaced by 2-aminoisobutyric acid and arginine respectively. Amino acid substitution at position 8 prevents chemical breakdown by an enzyme dipeptidyl peptidase-4. In addition, lysine at position 26 is in its derivative form (acylated with stearic diacid). Acylation with a spacer and C-18 fatty diacid chain increases the drug binding to blood protein (albumin), which enables longer presence in the blood circulation. Its half-life in the blood is about 7 days (165–184 hours), therefore, once-weekly injection is enough.
History.
Semaglutide was developed in 2012, by a team of researchers at Novo Nordisk as a longer-acting alternative to liraglutide. It was given the brand name Ozempic. Clinical trials were started in 2015, and phase III was completed in 2016.
Researchers at the University of Leeds and Novo Nordisk reported in 2017, that it can also be used for the treatment of obesity. It reduces hunger, food craving and body fat. A Phase 3 Randomized Controlled Trial found that once-weekly injection of 2.4 mg of the drug resulted in an average change of −14.9% body weight at 68 weeks compared to −2.4% for the placebo.
The US FDA New Drug Application (NDA) was filed in December 2016, and in October 2017, the FDA Advisory Committee voted 16–0 in favor. Approval was announced in December 2017. It can be used as both injection-type or oral-type drug. The marketing authorization in the European Union was granted in February 2018. The Japanese Ministry of Health, Labour and Welfare announced approval on 23 March 2018. Health Canada issued approval on 4 January 2018.
Semaglutide was approved for medical use in Australia in August 2019.


Biotechnology and Bioprocess Engineering

Biotechnology and Bioprocess Engineering is a peer-reviewed bimonthly scientific journal published by Springer Science+Business Media on behalf of the Korean Society for Biotechnology and Bioengineering. "Biotechnology and Bioprocess Engineering" covers all aspects of biotechnology and bioengineering. The editor-in-chief of the journal is Jong Won Yun (Daegu University) and Sang Yup Lee (KAIST). The founding editors-in-chief were Cha-Yong Choi (Seoul National University), Ho Nam Chang (Korea Advanced Institute of Science and Technology), and Sun Bok Lee (POSTECH).
Abstracting and indexing.
The journal is abstracted and indexed in EMBASE, EMBiology, Food Science and Technology Abstracts, IBIDS, International Abstracts in Operations Research, Science Citation Index Expanded, and Scopus. According to the "Journal Citation Reports", its 2019 impact factor is 2.213.


Exon trapping

Exon trapping is a molecular biology technique to identify potential exons in a fragment of eukaryote DNA of unknown intron-exon structure. This is done to determine if the fragment is part of an expressed gene.
The genomic fragment is inserted into the intron of a 'splicing vector' consisting of a known exon - intron - exon sequence of DNA, and the vector is then inserted into an eukaryotic cell. If the fragment does not contain exons (i.e., consists solely of intron DNA), it will be spliced out together with the vector's original intron. On the other hand, if exons are contained, they will be part of the mature mRNA after transcription (with all intron material removed). The presence of 'trapped exons' can be detected by an increase in size of the mRNA, or through RT-PCR to amplify the DNA of interest.
The technique has largely been supplanted by the approach of sequencing cDNA generated from mRNA and then using bioinformatics tools such as NCBI's BLAST server to determine the source of the sequence, thereby identifying the appropriate exon-intron splice sites.


PF-04479745

PF-04479745 is a research ligand developed by Pfizer. It is related to lorcaserin, and acts as a potent and selective agonist for the 5-HT2C receptor, with lower affinity and antagonist action at the related 5-HT2A and 5-HT2B receptor subtypes.


NSP5 (rotavirus)

NSP5 (nonstructural protein 5) encoded by genome segment 11 of group A rotaviruses. In virus-infected cells NSP5 accumulates in the viroplasms. NSP5 has been shown to be autophosphorylated.
Interaction of NSP5 with NSP2 was also demonstrated. In rotavirus-infected cells, the non-structural proteins NSP5 and NSP2 localize in complexes called viroplasms, where replication and assembly occur and they can drive the formation of viroplasm-like structures in the absence of other rotaviral proteins and rotavirus replication.
There is no atomic-resolution structure of NSP5 determined as of June 2019. However, the low resolution three-dimensional structure of the NSP2-NSP5 assembly has been observed by cryo-EM. NSP5 occupies the same site as RNA when binding to NSP2. The EM data from this 2006 study has not been published.


Ersentilide

Ersentilide is a beta adrenergic receptor antagonist.


(E,E)-2,4-Decadienal

("E","E")-2,4-Decadienal is an aromatic substance found in butter, cooked beef, fish, potato chips, roasted peanut, buckwheat and wheat bread crumb.
In an isolated state, it smells of deep fat flavor, characteristic of chicken aroma (at 10ppm). At lower concentration, it has the odor of citrus, orange or grapefruit.
It might be carcinogenic. It has been used as aroma in the EU, but use restrictions apply until the required data have been submitted.


Nitrite reductase

Nitrite reductase refers to any of several classes of enzymes that catalyze the reduction of nitrite. There are two classes of NIR's. A multi haem enzyme reduces NO2− to a variety of products. Copper containing enzymes carry out a single electron transfer to produce nitric oxide.
Iron based.
There are several types of iron based enzymes. Cytochrome cd1, or "Pseudomonas" cytochrome oxidase contains two c and two d type hemes with two polypeptide chains. Different forms of this reductase catalyze the formation of nitric oxide or nitrous oxide. A version of this compound was originally called [Ferrocytochrome c-551:oxidoreductase]. It was initially considered an oxidase. It catalyzes the reduction of NO2− to NO. This tetraheme enzyme has two subunits, each containing a c-type and a d-type heme. The reduced d hemes bind nitrite and convert it to product.
Cytochrome c nitrite reductase (ccNIR) is a multiheme enzyme that converts nitrite to ammonia on each active site. The active site iron is bound to a protoporphyrin IX ring that is covalently linked to the enzyme's proteins.
Proposed mechanism.
The ccNIR protein uses six electrons and seven hydrogens to reduce nitrite to ammonia. The active site of the enzyme contains an iron in a +2 oxidation state. The oxidation level allows nitrite to bond more strongly than to the +3 state due to increased pi backbonding. This electronic effect transfers electron density into the nitrite antibonding orbital between nitrogen and oxygen. The occupation of the LUMO decreases the strength of the N-O bond. A second electronic effect is the hydrogen bonding of both oxygens to nearby amino acids. These acids are often arginine and Histidine. The interactions lengthen the N-O bonds and facilitate cleavage of an oxygen from nitrogen.
The Fe-NO bond is linear and has six shared valence electrons. This is not a stable state for an Fe-NO bond. However, a bent seven electron configuration is too stable to undergo further reaction without considerable energy input. To compensate for this barrier, two rapid, consecutive, single electron reductions form an eight electron complex. The electron transfer occurs before a shift in geometry from a linear to bent geometry.
Two protonations of the nitrogen lead to an increased N-O bond distance. The resulting intermediate is a hydroxylamine. further protonation of the hydroxylamine leads to the breakage of the N-O bond to form water. The oxidation of iron from Fe(II) to Fe(III), coupled with a further protonation of nitrogen leads to the release of ammonia.
Copper based.
To date, there have been several types of Copper Nitrite Reductases discovered. These CuNIR are found in many different fungi and bacteria; for example, the bacterial genera "Pseudomonas", "Bordetella", "Alcaligenes", and "Achromobacter" all contain CuNIR. What is common to all CuNIR is the presence of at least one type 1 copper center in the protein. These centers are similar to Azurin in their bonding structure. Each type 1 Cu is strongly bonded to a thiolate sulfur from a cysteine, two imidazole nitrogens from different Histidine residues, and a sulfur atom of an axial Methionine ligand. This induces a distorted tetrahedral molecular geometry.
The cysteine ligated to the type 1 Cu center is located directly next to a Histidine in the primary structure of the amino acids. This Histidine is bound to the Type 2 Cu center responsible for binding and reducing nitrite. This Cys-His bridge plays an important role in facilitating rapid electron transfer from the type 1 center to the type 2.
Proposed mechanism.
The type 2 copper center of a copper nitrite reductase is the active site of the enzyme. The Cu is bound by nitrogens of two Histidines from one monomer, and bound by one Histidine from another monomer; the Cys-His bridge to the type 1 Cu. This gives the molecule a distorted tetrahedral geometry. In the resting state, the Cu is also binding a water molecule that is displaced by nitrite.
As nitrite displaces water, Cu is bound by both oxygens in a bidentate fashion. A nearby Aspartic acid residue hydrogen bonds to one of the newly formed oxygen ligands. An incoming electron reduces the Cu from oxidation state (II) to (I). This change facilitates a shift in nitrite binding so that the nitrogen is bound to Cu, and one oxygen has an extended bond length due to hydrogen bonding. A second hydrogen bond forms from Histidine or a nearby water molecule and leads to the cleavage of the N-O bond. The Cu is now five coordinate bonded to nitric oxide and water. Nitric oxide is released as Cu is oxidized to state (II) and returns to the resting configuration.
Assimilatory.
Assimilatory nitrate reductase is an enzyme of the assimilative metabolism involved in reduction of nitrate to nitrite. The nitrite is immediately reduced to ammonia (probably via hydroxylamine) by the activity of nitrite reductase.
The term assimilatory refers to the fact that the product of the enzymatic activity remains in the organism. In this case, the product is ammonia which has an inhibitive effect on assimilatory nitrate reductase, thus ensuring that the organism produces the ammonia according to its requirements.


William J. Rutter

William J. Rutter (born August 28, 1928) is an American biochemist who cofounded the early biotechnology company Chiron Corporation together with Edward Penhoet and Pablo DT Valenzuela. As Chairman of the Department of Biochemistry and Biophysics of the University of California, San Francisco, Rutter helped establish that department as a leader in the academic side of the biotechnology during the San Francisco Bay Area biotech boom of the 1980s.
Rutter spent a short time in the United States Navy and one year Brigham Young University, before completing a B.A. (1949) in biochemistry at Harvard University. He earned an M.S. (1950) from University of Utah and a PhD (1952) from the University of Illinois with a dissertation on galactosemia. Between 1952 and 1968, Rutter held positions at the University of Wisconsin, the Karolinska Institutet, University of Illinois, Stanford University, and University of Washington. In 1969, he moved to the University of California, San Francisco (UCSF), where he led the Department of Biochemistry and Biophysics until 1982. From 1983 until 1989, Rutter was Director of the Hormone Research Institute at UCSF.
In 1996, Rutter won the 2nd Annual Heinz Awards in Technology, the Economy and Employment and in 2003, he received the Biotechnology Heritage Award, from the Biotechnology Industry Organization (BIO) and the Chemical Heritage Foundation.


CCR2

C-C chemokine receptor type 2 (CCR2 or CD192 (cluster of differentiation 192) is a protein that in humans is encoded by the "CCR2" gene. CCR2 is a CC chemokine receptor.
Gene.
This CCR2 gene is located in the chemokine receptor gene cluster region. Two alternatively spliced transcript variants are expressed by the gene.
Function.
This gene encodes two isoforms of a receptor for monocyte chemoattractant protein-1 (CCL2), a chemokine which specifically mediates monocyte chemotaxis. Monocyte chemoattractant protein-1 is involved in monocyte infiltration in inflammatory diseases such as rheumatoid arthritis as well as in the inflammatory response against tumors. The receptors encoded by this gene mediate agonist-dependent calcium mobilization and inhibition of adenylyl cyclase.
Animal studies.
Alzheimer.
"CCR2" deficient mice have been shown to develop an accelerated Alzheimer's-like pathology in comparison to wild type mice. This is not the first time that immune function and inflammation have been linked to age-related cognitive decline (i.e. dementia).
Obesity.
Within the fat (adipose) tissue of CCR2 deficient mice, there is an increased number of eosinophils, greater alternative macrophage activation, and a propensity towards type 2 cytokine expression. Furthermore, this effect was exaggerated when the mice became obese from a high fat diet.
Myocardial Infarct.
CCR2 surface expression on blood monocytes changes in a time-of-day–dependent manner (being higher at the beginning of the active phase) and affects monocytes recruitment in tissues including the heart. As a consequence when an acute ischemic event happens during the active phase, monocytes are more susceptible to invade the heart. An excessive monocytes infiltration generates higher inflammation and increases the risk of heart failure.
Clinical significance.
In an observational study of gene expression in blood leukocytes in humans, Harries "et al." found evidence of a relationship between expression of "CCR2" and cognitive function (assessed using the mini-mental state examination, MMSE). Higher "CCR2" expression was associated with worse performance on the MMSE assessment of cognitive function. The same study found that "CCR2" expression was also associated with cognitive decline over 9-years in a sub-analysis on inflammatory related transcripts only. Harries "et al." suggest that "CCR2" signaling may have a direct role in human cognition, partly because expression of "CCR2" was associated with the ApoE haplotype (previously associated with Alzheimer's disease), but also because "CCL2" is expressed at high concentrations in macrophages found in atherosclerotic plaques and in brain microglia. The difference in observations between mice ("CCR2" depletion causes cognitive decline) and humans (higher "CCR2" associated with lower cognitive function) could be due to increased "demand" for macrophage activation during cognitive decline, associated with increased β-amyloid deposition (a core feature of Alzheimer's disease progression).


P.N.03

 short for Product Number Three, is a 2003 science-fiction third-person shooter video game developed by Capcom Production Studio 4 and published by Capcom for the GameCube. Set in a science-fictional space colony compound, the game follows a female mercenary fighting against berserk robots. It was directed by Shinji Mikami. To avoid similarities with "Devil May Cry" and "Resident Evil", the game designers emphasized defensive and rhythmic maneuvers, and a "delicate" "feminine" game world.
The game was developed on a tight schedule in an attempt to offset Capcom's poor fiscal year. However, it received mixed reviews and failed commercially. Several critics found the game repetitive and unintuitive. They were divided on its gameplay mechanics: some disliked its controls—specifically the inability to move and shoot simultaneously—while others compared it favorably to golden age arcade games. Mikami repurposed ideas from "P.N.03" for the game "Vanquish".
Gameplay and plot.
The player controls Vanessa Z. Schneider, a freelance mercenary working on colonized planets. She is contracted by a mysterious client to destroy Computerized Armament Management System (CAMS) robots that have gone berserk. Robots of this type had been responsible for killing Vanessa's parents. To combat the CAMS, Vanessa wears "Aegis suits", powered exoskeletons that allow her to fire energy beams from her palms and to perform powerful attacks called "energy drives". It depletes the energy meter, grants temporary invulnerability to Vanessa and damages multiple enemies. At checkpoints, the player uses points earned from defeated robots to purchase Aegis suits, suit upgrades and energy drives. The player may play "trial missions" (randomly generated levels) between missions to score extra points.
The CAMS robots attack in set patterns, and their next moves are indicated by visual and aural cues. The player avoids these attacks via evasive spins, rolls and other maneuvers. Vanessa cannot move while attacking, and so the player must often use the environment as cover. Defeated enemies sometimes drop items that replenish health and energy or that trigger a combo timer, which multiplies the points earned from enemies destroyed consecutively. Each mission takes place in a series of rooms that contain a fixed number of enemies and may include a boss robot. At the end of a mission, the player receives a score based on the number of rooms explored and enemies destroyed.
Late in the game, Vanessa discovers a clone of herself in a CAMS facility but is forced to abandon her when the building's self-destruct sequence is initiated. After destroying the CAMS central core, Vanessa encounters a digital projection of the client, whose appearance is identical to her own. She speculates that she is a clone of the client, but the client counters that none of their memories may be real. Vanessa debates whether to continue her work as a mercenary.
Development.
In November 2002, Capcom announced the Capcom Five, a group of video games developed by Capcom Production Studio 4 and overseen by "Resident Evil" creator Shinji Mikami. Among these titles was "P.N.03", directed by Mikami. Capcom conceived the Capcom Five to bring new intellectual property to the industry, which the company viewed at the time as stagnant. According to Hiroyuki Kobayashi, the "P.N.03" team's goal was to make a game both "fun to watch and fun to play". The team focused on the game's audiovisuals, action and speed, and they tried to balance the "tension experienced on the battlefield and the exhilaration of taking out the enemies". Mikami wanted "P.N.03" to evoke the same feelings as classic Nintendo games. While the staff felt the background graphics were important, they prioritized excitement in the game world.
During the planning stage, "P.N.03" was originally a wargame referred to as the "robot war game". Five days into development, the staff created a preliminary demonstration of the graphics. Mikami was unsatisfied with this prototype and decided to rework the project as a third-person shooting game. The director wanted to name the game "Jaguar" to reflect Vanessa's cat-like agility. However, other staff members disliked the name: some claimed that it failed to describe the game, while others thought that "White Jaguar" was a better title. Mikami chose "P.N.03" as a hint to the game's plot.
Early coverage of "P.N.03" by GameSpot and IGN highlighted the game's acrobatics and shooting. "P.N.03"s gameplay was initially similar to that of "Devil May Cry": Vanessa attacked with two pistols and performed acrobatic moves like Dante. Feeling that it resembled "Devil May Cry" too closely, Mikami altered the game to reward players for performing defensive moves. Mikami wanted Vanessa to use guns, but the developers couldn't complete the animated graphics for weapons in time for release. The guns were replaced with energy bolts fired from the character. In an effort to meet yearly sales goals, Capcom developed "P.N.03" quickly and released it in March 2003. Many of Capcom's releases for the year had underperformed. "Viewtiful Joe", another Capcom Five title, was delayed into the next fiscal year to decrease the possible sales that had to be offset. After leaving Capcom, Mikami later incorporated ideas he intended for "P.N.03" into the 2010 game "Vanquish".
Kobayashi aimed to avoid conventions established in "Resident Evil", such as that series' dark, masculine world. With "P.N.03", Kobayashi wanted to portray a "white" world with "feminine, delicate lines". To that end, the staff applied a minimalist approach to the visuals, used "fine drawn lines", and emphasized "visibility and creativity" in the game world. To maintain the game's "delicate image", the developers tried to make the visuals easy to view and understand. The team crafted the gameplay to avoid "button mashing"; instead, players were meant to observe enemy attack patterns and think before acting. Mechanical designer Shou Sakai tried to craft "things that would stick in [the player's] mind". Because of the tight development schedule, 3D models had to be created immediately after their designs were completed. Sakai described the lack of time as the "toughest part" of the process.
Mikami was indifferent to the main character's gender. During the early creation process, he left the decision to his staff, who ultimately chose a female character. Because "P.N.03" takes place in a space colony, Mikami wanted Vanessa's country of origin to be ambiguous. To that end, she was given a combination of French, German, and English names. Vanessa's movements were animated freehand, without motion capture technology. The designers posed Vanessa crouched on her hands and knees to resemble a jaguar, as an homage to the "Jaguar" title. The staff integrated rhythmic motions to her maneuvers to emulate dancing and to make her appear stylish. Kobayashi wanted Vanessa to be a "cool and sexy mercenary with a tough exterior that hides her dark past." In retrospect, he was proud of the character's style and movements.
Release.
The Capcom Five were first announced as games exclusive to the GameCube; however, Capcom later ported most of the titles to other consoles. At the end of 2002, the company confirmed that the game would be released in 2003 on GameCube. In January 2003, the company slated the game for a March release in Japan, but retracted the exclusivity announcement. At a press conference prior to the 2003 Electronic Entertainment Expo, however, Capcom reaffirmed that "P.N.03" would be available only for the GameCube. In the end, "P.N.03" was the only Capcom Five title to remain exclusive to the system.
"P.N.03" was the first of the Capcom Five to be released. Before the game's debut in Japan, Capcom distributed playable demos to stores and released screenshots to the media. Upon the game's release, Capcom shipped 25,000 copies to Japanese retailers. Under 11,000 units were sold, which made the game Japan's 26th best-selling title during the last week of March 2003. These low sales failed to help Capcom meet its yearly sales goals. The North American localization was announced in July 2003. Few aspects of the game were changed for the North American market. Capcom advertised a free T-shirt as a pre-order incentive in North America. Promotional sunglasses modeled after Vanessa's were also released.
Reception.
"P.N.03" failed commercially and received mixed reviews, with scores of 64% and 63 on review aggregate websites GameRankings and Metacritic, respectively. "Electronic Gaming Monthly"s three reviewers characterized the game as shallow, repetitive and devoid of plot and character development. Mark MacDonald of the magazine criticized Vanessa's inability to move and shoot at the same time. By contrast, the reviewer for "Edge" wrote, ""P.N.03" may be rather short and its premise simple, but grace under fire has rarely been done better." The writer favorably compared its gameplay to that of "Space Invaders", in that the game "rewards skill above all else and mastery brings huge satisfaction". The reviewer cited Vanessa as one of the game's high points. In a 2009 retrospective review, the "Edge" magazine staff echoed its previous praise of "P.N.03" but acknowledged the game's awkward control mechanics. The staff commented that Vanessa's potentially fluid movement is difficult to execute, but that the challenge of mastering the control scheme is part of the game's charm.
"GamePro"s Mike Weigand called the game "a long, intense, thumb-busting shooter that tests trigger fingers and patience levels." He found the environments and enemies repetitive, but he cited "strong aesthetics" and "a slick reward system" as redemptive features. Weigand summarized that, while the game lacks depth, it may be recommended to "those who crave a shoot-em-up with old-school inflections". Paul Byrnes of "GMR" found "P.N.03" to be a "boring and repetitive" missed opportunity. He felt that it lacked flow, thanks in large part to Vanessa's "clumsily staccato" movements and inability to move and shoot simultaneously. "Game Informer"s Andrew Reiner wrote that "P.N.03"s setting, animations and protagonist give it an "undeniable allure" of freshness and originality. However, he disliked the game's control system and wrote that "blowing away robots gets old rather quickly". Reiner called "P.N.03"s length, which he estimated to be four hours, its "most disappointing aspect".
Greg Kasavin of GameSpot wrote that "you'll almost certainly be unimpressed with the repetitive and cumbersome action at the heart of ["P.N.03"]." He disliked the game's separation into brief, discrete sections; and, while he saw the game's design as a reference to that of older games, he found that "P.N.03" lacked the "extremely precise controls and smooth, colorful graphics" of the titles that inspired it. Kasavin summarized it as "a short, uninspired game that's yet another would-be imitator of Capcom's own "Devil May Cry"." Matt Casamassina of IGN wrote that players will ""want" to like" the game, but that its "design flaws and sloppy execution" are impossible to overlook. He found it to be shallow and repetitive, and he wrote that its fast pace and "unresponsive control setup" combine to place the game "at war with itself." However, he believed that "P.N.03" is "not a disaster—merely a disappointment", and that it sits "solidly in average country." In a 2012 retrospective from Eurogamer, Chris Schilling called the game the outcast of Shinji Mikami's work, criticizing its plot and calling its control scheme its "biggest obstacle". He later added "if at times its appeal can be hard to pin down, that doesn't mean it should be so easily forgotten", mentioning the satisfaction of beating the bosses in the game.


Dimethylglycine dehydrogenase

In enzymology, a dimethylglycine dehydrogenase () is an enzyme that catalyzes the chemical reaction
The 3 substrates of this enzyme are N,N-dimethylglycine, acceptor, and H2O, whereas its 3 products are sarcosine, formaldehyde, 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 N,N-dimethylglycine:acceptor oxidoreductase (demethylating). Other names in common use include N,N-dimethylglycine oxidase, and N,N-dimethylglycine:(acceptor) oxidoreductase (demethylating). This enzyme participates in glycine, serine and threonine metabolism. It employs one cofactor, FAD.


Annette Herscovics

Annette Herscovics (1938–2008) was a scientist at McGill University, a Fellow of the Royal Society of Canada, and a pioneer in the field of glycobiology.
Personal life.
Herscovics was born in Paris, France, the daughter of Polish Jews. She survived the Holocaust as a hidden child in Nazi-occupied France. After immigrating to Canada, she obtained a PhD in biochemistry at McGill University in Montreal in 1963.
Herscovics died of cancer on September 6, 2008.
Career.
Herscovics worked in the Department of Anatomy at McGill from 1967–1971, during which she made several important discoveries in glycobiology. She discovered in 1969 that thyroglobulin undergoes carbohydrate modifications, part of a class of proteins known as glycoproteins.
After completing her post-doctoral work at McGill, Herscovics moved to Harvard Medical School in 1971, where she remained until 1981. During this time she published more than 20 original papers in her field.
In 1981, Herscovics returned to McGill University as an associate professor in the McGill Cancer Centre. She was appointed a professor in the Departments of Medicine and Biochemistry in 1987, and a professor in the Department of Oncology in 1992. During this time she made several other important discoveries, including how the carbohydrate modifications are relevant to disease, including cancer.
Herscovics was elected as a Fellow of the Royal Society of Canada in 1998. She has published 110 peer-reviewed papers during her academic career.


CAAT box

In molecular biology, a CCAAT box (also sometimes abbreviated a CAAT box or CAT box) is a distinct pattern of nucleotides with GGCCAATCT consensus sequence that occur upstream by 60–100 bases to the initial transcription site. The CAAT box signals the binding site for the RNA transcription factor, and is typically accompanied by a conserved consensus sequence. It is an invariant DNA sequence at about minus 70 base pairs from the origin of transcription in many eukaryotic promoters. Genes that have this element seem to require it for the gene to be transcribed in sufficient quantities. It is frequently absent from genes that encode proteins used in virtually all cells. This box along with the GC box is known for binding general transcription factors. Both of these consensus sequences belong to the regulatory promoter. Full gene expression occurs when transcription activator proteins bind to each module within the regulatory promoter. Protein specific binding is required for the CCAAT box activation. These proteins are known as CCAAT box binding proteins/CCAAT box binding factors.
A CCAAT box is a feature frequently found before eukaryote coding regions, but is not found in prokaryotes.
Consensus sequence.
In the direction of transcription of the template strand, the consensus sequence, or the calculated order of the most frequent residues, for the CAAT box was 3'-TG ATTGG (T/C)(T/C)(A/G)-5'. The use of parentheses denotes that either base is present, but it is not specified as to their relative frequencies. For example, "(T/C)" would mean that either thymine or cytosine are preferentially selected for. Within metazoa (animal kingdom), the core binding factor (CBF)-DNA complex retains a high degree of conservation within the CCAAT binding motif, as well as the sequences flanking this pentameric motif. The CCAAT motif in plants (spinach was used in an experiment) differs slightly from metazoa in that it is actually a CAAT binding motif; the promoter lacks one of the two C residues from the pentameric motif, and the artificial addition of the second C has no significant effects on binding activity. Some sequences lack the CAAT-box completely. Secondly, the surrounding nucleotides in plants do not match the consensus sequence above determined by Bi "et al."
Core promoter.
The CAAT box is what is known as a core promoter, also known as the basal promoter or simply the promoter, is a region of DNA that initiates transcription of a particular gene. This region, in particular for the CAAT box, is located about 60–100 bases upstream (towards the 5' end), however no less than 27 base pairs away, from the initial transcription site or a eukaryote gene in which a complex of general transcription factors bind with RNA polymerase II prior to the initiation of transcription. It is essential to the transcription that these core binding factors (also referred to as nuclear factor Y or NF-Y) are able to bind to the CCAAT motif. Experiments in many laboratories have shown that mutations to the CCAAT motif that cause a loss of CBF binding also decreases transcriptional activity in these promoters, suggesting that CBF-CCAAT complexes are essential for optimum transcriptional activity.
Binding.
In an experiment done with core binding factors (CBF) and DNA complexes, researchers were able to determine the preferential sequences of the promoter in a region over and immediately adjacent to the CAAT box, and two regions on either side of the CAAT box. By using PCR-mediated random binding selection process, researchers were able to show that the sequence "3' - (T/C)G ATTGG (T/C)(T/C)(A/G) - 5'" immediately flanking the ATTGG region (CCAAT in the complementary strand) was preferentially selected on the coding strand (opposite of the template strand). This was shown using an oligonucleotide sequence (R1) which contained 27 random nucleotides, flanked by a defined 20 nucleotide sequence on each side. While no single nucleotide was selected in every clone on either side of the ATTGG motif (CCAAT in the complementary strand), there were several nucleotides in positions selected with high frequency. Most notably from the sequence above was the G residue towards the 5' end of the ATTGG. The other residues also listed were notable, but there is a split between two residues. This same experiment also yielded the same sequence as shown above when using a different oligonucleotide (R2) that contained an ATTGG core and flanked by 12 5' random nucleotides and 10 3' random nucleotides. Both these sequences are very similar and confirmed in multiple experiments. For sequences that flanked the ATTGG motif with two adenine residues (AA) on its 5' end and G(A/G) on its 3' end, seems to have inhibited formation of the CBF-DNA complex and subsequently occurred in only 1% of the promoter sequences. In another experiment performed with the major late promoter (MLP) of adenoviruses from a variety of host species, it was shown that the mutation of the CAAT box and CCAAT sequence, which is thought to play a pivotal role in the (MLP) of subgroup C human adenoviruses, in species with a deficient CAAT sequence. The transcription initiation at mutant MLP species was significantly reduced compared with that of the wild type or species in which there was a CAAT mutant. The failure to restore the normally functional adenoviruses, exhibited by a CAAT box, is consistent with the idea that the CAAT box plays a vital role in the adenovirus MLP and is preferred over other transcriptional elements.
CCAAT in plants.
These core binding factors, or nuclear factors (NF-Y), are composed of three subunits – NF-YA, NF-YB, and NF-YC. Whereas in animals each NF-Y subunit is encoded by a single gene, there has been a diversification in plants in both structure and function. Families of NF-Y consist of between eight and 39 members per subunit. A large reason for this diversification is because of gene duplications and tandem duplications, which have helped contribute to the larger family sizes of NF-Y compared to the single encoded animal nuclear factors. Each subunit contains an evolutionarily conserved part – the C-terminal of NF-YA, the central part of NF-YB, and the N-terminal of NF-YC, greater than 70% of these across species remains conserved. Neighboring regions however are generally not conserved.
NF-YA subunit.
The NF-YA family encodes transcription factors that are variable in length (between 207–347 amino acids for "M. truncatula"). The NF-YA proteins are generally characterized by two domains that are strongly conserved in all higher eukaryotes investigated to date. The first domain (A1) contains 20 amino acids that forms an alpha helix that appears significant in its interactions with NF-YB and NF-YC. The second domain (A2) is adjacent to the A1 domain by a conserved linker sequence is a sequence of 21 amino acids vital in the specific DNA to CCAAT box binding. The A1 and A2 domains are conserved towards the C-terminus of mammals, but occupy a more central region in plant NF-YA subunits. In plants, the NF-YA subunit has evolved to regulate the development of a facultative root organ only present in leguminous plants and shown to be expressed in root tissue. It was shown to have drought-resistant-like properties, becoming upregulated during drought stress in the roots and leaves of "Arabidopsis". NF-YA mutants have shown a loss of function and a hypersensitivity to drought-like conditions, and contrastly, overexpression of NF-YA has resulted in drought resistance.
NF-YB subunit.
The NF-YB family is, similar to the NF-YA subunit, variable in length, however, on average much smaller than the NF-YA subunit (90–240 amino acids in "M. truncatula"). They have been characterized with a structure and amino acid composition similar to the histone fold motif (HFM). This is composed of three alpha-helices separated by two beta strand-loop domains. Similar to NF-YA, NF-YB has been shown to also improve drought resistance when overexpressed and also the promotion of flowering in "Arabidopsis".
NF-YC subunit.
The NF-YC proteins are an intermediate size between that of NF-YA and NF-YB proteins (117–292 amino acids in "M. truncatula") and also contain the HFM that is prevalent in NF-YB proteins. It has also been shown to be involved in flowering time in certain plants (overexpression leads to earlier flowering) where its influence is potentially regulated by the binding of the protein CONSTANS (CO) to the NF-YC subunit.
NF-Y complexes.
Because of the evolutionary change in NF-Y encoding genes in plants, they subsequently have a large range of potential trimeric complexes. For example, in "Arabidopsis", 36 NF-Y transcription factor subunits (including 10 NF-YA, 13 NF-YB, and 13 NF-YC subunits) have been identified and which could theoretically form 1690 unique complexes (which contains one of each type of subunit). This number, of course is higher than what actually happens since some subunits have specific binding patterns. Functional analyses on NF-Y encoding genes in plants have shown, as a result of their evolutionary diversification relative to their animal counterparts, have acquired diverse specific functions, such as embryo development, flowering time control, ER-stress, drought stress, and nodule and root development. This may only be a small portion of their capabilities, since the number of theoretically combinations of NF-Y complexes is so large and only a small portion can actually be created (less than 10% of all possible interactions were confirmed in both directions in yeast).
CCAAT enhancer binding proteins (C/EBPs).
Another aspect of the CCAAT binding motif is the CCAAT/enhancer binding proteins (C/EBPs). They are a group of transcription factors of 6 members (α-ζ), which are highly conserved and bind to the CCAAT motif. While research on these binding proteins is relatively recent, their function has been shown to have vital roles in cellular proliferation and differentiation, metabolism, inflammation, and immunity in various cells, but specifically hepatocytes, adipocytes, and hematopoietic cells. For example, in adipocytes, this has been shown in a variety of experiments with mice: ectopic expression of these C/EBPs (C/EBPα and C/EBPβ) were able to initiate the differentiation programs of the cell, even in the absence of adipogenic hormones, or the differentiation of preadipocytes to adipocytes (or fat cells). In addition, an overabundance of these C/EBPs (specifically, C/EBPδ) causes an accelerated response. And furthermore, in cells lacking C/EBP or in C/EBP-deficient mice, both are unable to undergo adipogenesis. This results in the mice dying from hypoglycemia, or the reduced lipid accumulation in adipose tissue. The C/EBPs follow a general basic-leucine zipper (bZIP) domain at the C-terminus and are able to form dimers with other C/EBPs or other transcription factors. This dimerization allows the C/EBPs to bind specifically to DNA through a palindromic sequence in the major groove of DNA. They are regulated through various means, including hormones, mitogens, cytokines, nutrients, and other various factors.


Insulin-like growth factor II IRES

The insulin-like growth factor II (IGF-II) internal ribosome entry site IRES is found in the 5' UTR of IGF-II leader 2 mRNA. This RNA element allows cap-independent translation of the mRNA and it is thought that this family may facilitate a continuous IGF-II production in rapidly dividing cells during development. Ribosomal scanning on human insulin-like growth factor II (IGF-II) is hard to comprehend due to one open reading frame and the ability for the hormone to fold into a stable structure.


GDP-D-glucose phosphorylase

GDP-D-glucose phosphorylase () is an enzyme with systematic name "GDP:alpha-D-glucose 1-phosphate guanylyltransferase". This enzyme catalyses the following chemical reaction
The enzyme may be involved in prevention of misincorporation of glucose in place of mannose residues into glycoconjugates.


CXC chemokine receptors

CXC chemokine receptors are integral membrane proteins that specifically bind and respond to cytokines of the CXC chemokine family. They represent one subfamily of chemokine receptors, a large family of G protein-linked receptors that are known as seven transmembrane (7-TM) proteins, since they span the cell membrane seven times. There are currently six known CXC chemokine receptors in mammals, named CXCR1 through CXCR6.
CXCR1 and CXCR2.
CXCR1 and CXCR2 are closely related receptors that recognize CXC chemokines that possess an E-L-R amino acid motif immediately adjacent to their CXC motif. CXCL8 (otherwise known as interleukin-8) and CXCL6 can both bind CXCR1 in humans, while all other ELR-positive chemokines, such as CXCL1 to CXCL7 bind only CXCR2. They are both expressed on the surface of neutrophils in mammals.
CXCR3.
CXCR3 is expressed predominantly on T cells (T lymphocytes), and also on other lymphocytes [some B cells (B lymphocytes) and NK cells] and is highly induced following cell activation. There are two isoforms, CXCR3-A and CXCR3-B. It has three highly related ligands in mammals, CXCL9, CXCL10 and CXCL11.
CXCR4.
CXCR4 (also known as fusin) is the receptor for a chemokine known as CXCL12 (or SDF-1) and, as with CCR5, is utilized by HIV-1 to gain entry into target cells. This receptor has a wide cellular distribution, with expression on most immature and mature hematopoietic cell types (e.g. neutrophils, monocytes, T and B cells, dendritic cells, Langerhans cells and macrophages). In addition, CXCR4 can also be found on vascular endothelial cells and neuronal/nerve cells.
CXCR5.
The chemokine receptor CXCR5 is expressed on B cells and CD4+ Tfh cells and is involved in lymphocyte homing and the development of normal lymphoid tissue. Its principal ligand is CXCL13 (or BLC).
CXCR6.
CXCR6 was formerly called three different names (STRL33, BONZO, and TYMSTR) before being assigned CXCR6 based on its chromosomal location (within the chemokine receptor cluster on human chromosome 3p21) and its similarity to other chemokine receptors in its gene sequence. CXCR6 binds the ligand CXCL16. However, CXCR6 is more closely related in structure to CC chemokine receptors than to other CXC chemokine receptors.
History.
ACKR3 was originally called RDC-1 (an orphan receptor) but has since been shown to cause chemotaxis in T lymphocytes in response to CXCL12 (the ligand for CXCR4) prompting the renaming of this molecule as CXCR7. ACKR3 designation has been accepted by the IUIS/WHO Subcommittee on Chemokine Nomenclature. This receptor has also been identified on memory B cells.


Kelba quadeemae

Kelba quadeemae is an extinct species of ptolemaiidan mammal, the sole species of the family Kelbidae, known from the Lower Miocene of East Africa. The genus name "Kelba" derives from the Arabic (pronounced "kalb" or "kelb") meaning "dog", and the specific name "quadeemae" from the Arabic "quadeem", meaning "ancient". "Kelba" is only known from a partial skull and teeth, but estimated to have been (around 15 kg), similar in size to a coyote but more heavily built. The teeth are rather unspecialized, suggesting a wide and varied diet, and show wear suggesting its diet included abrasive material.


Pacomio

José Pacomio Poqui, better known as Pacomio, was a Chumash revolutionary, carpenter and "comisario de policia" (Police commissioner) of Monterey, who led one of the most significant Native American uprisings against the Spanish and Mexican rule of California, the Chumash Revolt of 1824. Pacomio's revolt against the Mexican-Spanish establishment was perhaps the most brutal in California's history, and grimly showed the Mexican government that California's indigenous population was not as servile as once thought.
Early life.
Pacomio Poqui was born on La Ranchería de Esniscue, Alta California around 1794 to Chumash parents, and was baptized and given the Spanish name José at Mission La Purisima in 1803. During his time as a young boy at Mission La Purisima, he proved to be extraordinarily intelligent, which won him the favor of the mission's Franciscan friars. The friars taught the young Chumash how to read and speak Spanish, which was rarely practiced by the Spaniards. Most priests did not bother to teach the new indigenous converts their Spanish language, focusing instead on teaching them the basic symbols and ideology of Catholicism. Pacomio's fluent understanding of Spanish would become invaluable later in life.
In August 1807, at the age of thirteen, Pacomio married the eleven-year-old neophyte Gordiana. The couple would live childless in Mission La Purisima for twelve more years, until Gordiana's unexpected death in 1819. At the same time, Pacomio trained to become a carpenter, working under the watchful eyes of Salvador Carabantes and, after 1811, master stonemason José Antonio Ramírez. In December 1812, Mission La Purisima was totally destroyed by an earthquake, and the craftsmen of the former mission were instructed to build a new version three miles to the north of the original site. Pacomio and other neophyte craftsmen, like Mariano Francisco, Sebastián Tomás and Juan Nepomunceno, worked hard on the arduous task. Upon Gordiana's death, Pacomio and Tomás traveled to Monterey to work on the Presidio. After his duties in the Presidio expired, Pacomio took a liking to the city, and settled there. In February 1820 he married the widow Eusebia María, and in December had a daughter, María de Jesús. However, in 1824, growing tensions between his people, the Chumash, and the new Mexican Government forced him to leave his family in Monterey. Little did Pacomio know how significant those tensions were to become.
Chumash Revolt of 1824.
When Mexico gained control over Spanish California in 1821, following its own independence from Spain, things seemed hopeless for the former Spanish colonists and soldiers. Not only had a former Spanish colony won independence and taken over their land, but under the orders of the unpopular King Ferdinand VII, the Spanish government had stopped their funding of all of the Spanish missions and presidios in California. This news was too much for the Spanish settlers to bear, and former Spanish soldiers began to take their anger out on the Native Californians. On February 21, 1824, a young Chumash boy from Mission La Purisima was severely beaten by a Mexican soldier when he was visiting a relative imprisoned inside the Mission Santa Inés guardhouse. This brutal act caused the Chumash neophytes in the mission to take up arms against the soldiers, attacking them with arrows and setting multiple buildings on fire. Two of the Chumash assailants were killed. After a heated battle with many wounded and the arrival Chumash reinforcements, the mission's priest and soldiers barricaded themselves inside a building, where they waited to be rescued until the next day, by a detachment of Mexican troops from the Presidio of Santa Barbara. The soldiers forced the Santa Inés rebels into the neophyte housing of the mission, which they promptly burnt down to flush the Chumash out. Some of the Chumash ran off to the two nearest missions, Santa Barbara and La Purisima, to inform their fellow Chumash of the revolt, and to join them. One messenger met with the "Wot" (leader) of the Chumash tribe, and informed him of the successful revolt against Mission Santa Inés. The Chumash leader ordered a call to arms among his people and more attacks on the surrounding missions, as well as the evacuation of all women and children to the mountains. The Chumash warriors armed themselves with bows and arrows and machetes, preparing for the inevitable conflict against the Mexican soldiers.
As the Chumash revolted against the Mexican soldiers inside Mission Santa Inés, a group of 2,000 Chumash warriors made a frontal assault on Mission La Purisima, capturing it and taking the soldiers, priests and civilians inside as prisoners. During the assault of the mission, one Chumash warrior was killed, and in retaliation, the rebels massacred four innocent travelers that they had captured during the battle. On February 24, The surviving soldiers and civilians inside La Purisima were allowed to travel back to the safety of Mission Santa Inés unharmed. Only one Franciscan, Father Antonio Rodríguez, stayed behind with the Chumash insurgents.
On February 22, the next day after the initial revolt, the Chumash occupiers were reinforced by the warriors fleeing from the skirmish at Santa Inés, as well as Chumash from Mission San Fernando Rey de España. The insurgents began to fortify La Purisima, erecting wooden palisades and cutting gun loops out of the mission's walls, arming themselves with the mission's muskets. Simultaneously, Mission Santa Barbara was also captured by another force of Chumash, who forced the mission's soldiers, clergy and civilians to retreat to the nearby Santa Barbara Presidio. Soon after the capture of Mission Santa Barbara, however, a small force of Mexican troops and priests arrived at the mission from the presidio, attempting to negotiate the surrender of the Santa Barbara rebels. Predictably, the Chumash refused, and a heated battle raged inside the mission for multiple hours, ending with two Chumash killed and three wounded, and four Mexican soldiers wounded. The Mexican detachment fled back to the presidio, while the Chumash defenders sacked the mission of its valuables and supplies, and retreated into the back country.
Upon hearing the news of the revolt at Santa Inés, as well as the uprisings at the surrounding missions, Pacomio was overjoyed. His people had finally resisted against the cruel subjugation that they had faced for many years. Pacomio left his family after the Santa Inés revolt, to join up with the Chumash at Mission La Purisima. He assumed command over the force of 400 Chumash there, and began to train them in the art of European warfare. Pacomio issued his warriors gunpowder and muskets, drilling his men endlessly with the intention that they would become proficient with the foreign weaponry. He also trained them to use the mission's two swivel guns, in order to bring heavier firepower against the well-trained Mexican Army.
Nearly a month after the initial revolt on February 21, 1824, a company of 100 Mexican soldiers, cavalrymen and priests, as well as a four-pound cannon, arrived at Mission La Purisima in the morning, intent on violently crushing the rebellion. The Mexican Governor of Alta California, Luis Antonio Argüello, had had enough chaos in his country's new territory, and had given the order for the Chumash rebellion to be quelled with bloodshed, if necessary. A bloody battle broke out at Mission La Purisima, with Pacomio bravely leading the insurgents against the Mexicans. The Chumash let loose precise musket volleys, which scattered the Mexican ranks, and torrents of lethal arrows rained down upon the Mexican soldiers and cavalry. The Mexicans charged the Chumash with their cavalrymen, armed with muskets, who provided support and covering fire for the infantry. The four-pound cannon opened fire on the insurgents, killing many and causing disorder among the Chumash ranks. Eventually, as the battle progressed, Pacomio ordered the mission's two swivel guns to fire upon the enemy. However, the Chumash manning the guns had very little experience with them, and the two swivel guns misfired, exploding in the gunners' faces, killing them instantly. Dismayed, Pacomio commanded his men to hold strong, despite the loss of the cannons. After a morning of intense fighting, a ceasefire was negotiated by Father Rodríguez. The casualties at the end of the battle were appalling: five Mexican soldiers were killed, as well as many wounded, while sixteen Chumash were killed and wounded. After the Battle of La Purisima, Pacomio and his remaining warriors surrendered. The captured La Purisima rebels were rounded up and taken to Monterey, where they were tried in court. Seven Chumash were convicted of the murder of the four travelers at Mission La Purisima, and were publicly executed by Hanging. The four leaders of the revolt, Pacomio, Mariano, Benito and Bernarde, were sentenced to ten years of chain gang labor (Benito and Bernarde managed to escape the Monterey prison, and fled into the mountains to join their kin). After two months of unrest, the bloodiest Native American uprising in Californian history was over.
Later life and death.
After serving his sentence of ten years of hard labor for his role in the Chumash Revolt of 1824, Pacomio was reunited with his family in Monterey in 1834. He continued with his skill in carpentry, making furniture for the Monterey Presidio and Mission La Purisima. Pacomio's business began to flourish, as Monterey's population was booming. The population of 1,600 in 1830 would increase to nearly 2,000 in 1845. At the age of thirteen, in 1833, María de Jesús married Gregorio, a Chumash neophyte who participated in the Revolt. In 1836, however, soon after moving into her father's house with her husband, María died in childbirth at the age of sixteen.
Despite the loss of his daughter, Pacomio was becoming an influential man in the Monterey community and local government. His contemporaries described him as "a well-educated neophyte, skillful carpenter and cabinet-maker" as well as "an intelligent citizen and member of the Ayuntamiento (city council) of Monterey." Ironically, the man who had led the Chumash in revolt in 1824 was offered the position of Monterey's "comisario de policia", or Police commissioner, in 1836, which Pacomio gladly accepted. Throughout the rest of his life in Monterey, Pacomio never lost touch with his native heritage and identity. Citizens walking past his house regularly observed him outside, singing "Chumash songs while dancing, dressed only in a breech-clout, his body painted red, white and black, and with feathers on his head." Unfortunately, Pacomio's life was a relatively short one, with him and wife both perishing from the Monterey Smallpox Epidemic of 1844. Monterey had lost a man whose exploits had changed California's history, and ultimately, America's history as well.


Infante Pedro Carlos of Spain and Portugal

Don Pedro Carlos (Pedro Carlos Antonio Rafael José Javier Francisco Juan Nepomuceno Tomás de Villanueva Marcos Marcelino Vicente Ferrer Raimundo; 18 June 1786 – 4 July 1812) was an Infante of Spain and Portugal.
Family.
Infante Pedro Carlos was a son of Infante Gabriel of Spain and Infanta Mariana Victoria of Portugal. His paternal grandfather was King Charles III of Spain and his maternal grandfather was King Peter III of Portugal. Pedro Carlos was the only surviving child of the couple and an orphan at the age of two.
Life.
His father, a very intelligent man, was King Charles III's favorite son, but he and his wife died in 1788 of smallpox. King Charles III also died the same year; his successor Charles IV of Spain sent the child away to Portugal, as requested by his grandmother, Queen Maria I of Portugal. Maria was worried about the Braganza succession and Pedro Carlos was then her only grandchild. He was raised by Maria, who created him an infante of Portugal.
Pedro Carlos had inherited a large fortune from his father and was welcomed in Portugal. In 1792 his grandmother was officially declared insane and her son John, uncle of Pedro Carlos, became Regent of Portugal.
In 1807, France invaded Portugal, and the royal family fled to Brazil, including Pedro Carlos. They left on 29 November on board the "Príncipe Real".
They arrived in Salvador de Bahia on 2 January 1808 and headed from there to Rio de Janeiro, where the infante lived in the São Cristovão Palace.
Marriage.
In Brazil Pedro Carlos was engaged to his cousin Maria Teresa, Princess of Beira, daughter of Carlota Joaquina of Spain and John VI of Portugal, and they married on 13 May 1810 in Rio de Janeiro.
The couple was very happy during their two years of marriage, after which Pedro Carlos became ill and died in Alto da Boa Vista on 4 July 1812.
The couple had one child:


MinD

The MinD protein is one of three proteins encoded by the "minB" operon and also a part of the ParA family of ATPases. It is required to generate pole to pole oscillations prior to bacterial cell division as a means of specifying the midzone of the cell. It is a peripheral membrane ATPase involved in plasmid partitioning.
History.
When first discovered in "E.coli", MinD was thought to associate with MinC and form a stable cap at each bacterial pole, thereby specifying the mid-zone of the cell by alleviating inhibitory pressures in that region. Through the use of live-cell imaging with GFP fusion proteins, Raskin and de Boer revealed a dynamic interaction of the Min proteins, demonstrating that MinC and MinD instead rapidly oscillate between the two poles in a non-static manner.
Function.
The ATPase activity of MinD is activated by MinE while in the presence of phospholipids, suggesting that the binding to the membrane induces a conformational change allowing it to become susceptible to MinE activation.
MinD activity is dependent on local MinD concentration, suggesting an oligomerization process
and cooperativity.
"In vitro" studies.
"In vitro" studies of the Min system are done on a two dimensional supporting lipid bilayer. Fluorescent labelling of MinD has revealed that it tends to form a membrane-bound carpet in MinE mutants (ATP cannot be limiting). Upon re-addition of MinE, the system becomes unstable and dynamic. Localized foci of increased MinE concentration result in subsequent MinD membrane detachment and through several iterations of this attachment and detachment process, the emergence of standing waves was observed. This process is indicative of the Min system’s ability to self-organize. Fluorescence studies reveal the formation of a focused wave-front.
Single molecule dynamics revealed that MinD dimerizes while membrane-bound, resulting in a stronger membrane association at the rear of the standing wave and giving rise to a diffusivity gradient. This observation explains the presence of focused bands of fluorescence in these standing wave studies.
Additional study of this system and its interacting molecular partners is required to fully characterize the min system and to understand the molecular dynamics.


N-methylphosphoethanolamine cytidylyltransferase

In enzymology, a N-methylphosphoethanolamine cytidylyltransferase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are CTP and N-methylethanolamine phosphate, whereas its two products are diphosphate and CDP-N-methylethanolamine.
This enzyme belongs to the family of transferases, specifically those transferring phosphorus-containing nucleotide groups (nucleotidyltransferases). The systematic name of this enzyme class is CTP:N-methylethanolamine-phosphate cytidylyltransferase. Other names in common use include monomethylethanolamine phosphate cytidylyltransferase, and CTP:P-MEA cytidylyltransferase.


Muscarinic acetylcholine receptor M4

The muscarinic acetylcholine receptor M4, also known as the cholinergic receptor, muscarinic 4 (CHRM4), is a protein that, in humans, is encoded by the "CHRM4" gene.
Function.
M4 muscarinic receptors are coupled to Gi/o heterotrimeric proteins.
They function as inhibitory autoreceptors for acetylcholine. Activation of M4 receptors inhibits acetylcholine release in the striatum. The M2 subtype of acetylcholine receptor functions similarly as an inhibitory autoreceptor to acetylcholine release, albeit functioning actively primarily in the hippocampus and cerebral cortex.
Muscarinic acetylcholine receptors possess a regulatory effect on dopaminergic neurotransmission. Activation of M4 receptors in the striatum inhibit D1-induced locomotor stimulation in mice. M4 receptor-deficient mice exhibit increased locomotor simulation in response to D1 agonists, amphetamine and cocaine. Neurotransmission in the striatum influences extrapyramidal motor control, thus alterations in M4 activity may contribute to conditions such as Parkinson's disease.


Leucine dehydrogenase

In enzymology, a leucine dehydrogenase () is an enzyme that catalyzes the chemical reaction
The 3 substrates of this enzyme are L-leucine, H2O, and NAD+, whereas its 4 products are 4-methyl-2-oxopentanoate, NH3, NADH, and H+.
This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH-NH2 group of donors with NAD+ or NADP+ as acceptor. The systematic name of this enzyme class is L-leucine:NAD+ oxidoreductase (deaminating). Other names in common use include L-leucine dehydrogenase, L-leucine:NAD+ oxidoreductase, deaminating, and LeuDH. This enzyme participates in valine, leucine and isoleucine degradation and valine, leucine and isoleucine biosynthesis.
Structural studies.
As of late 2007, only one structure has been solved for this class of enzymes, with the PDB accession code .


Phenylalanine 2-monooxygenase

In enzymology, a phenylalanine 2-monooxygenase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are L-phenylalanine and O2, whereas its 3 products are 2-phenylacetamide, 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-phenylalanine:oxygen 2-oxidoreductase (decarboxylating). Other names in common use include L-phenylalanine oxidase (deaminating and decarboxylating), and phenylalanine (deaminating, decarboxylating)oxidase. This enzyme participates in phenylalanine metabolism.


Allen County Public Library

The Allen County Public Library (ACPL) is a public library system located in Fort Wayne, Allen County, Indiana, United States. Founded in 1895 as the Fort Wayne Public Library, the library served residents with 3,606 books out of a single room in City Hall. Today the library system includes 14 library branches within Fort Wayne and throughout the county. The Main Library Branch in downtown Fort Wayne is home to the Fred J. Reynolds Historical Genealogy Department, which holds the largest public genealogy collection in the United States.
Timeline.
1895 – Library opened in City Hall on January 28 with 3,606 volumes.
1898 – Library Board purchased the Brackenridge Home at Wayne Street and Webster Street for $14,000.
1904 – Carnegie-funded library building opened.
1923 – Service to county residents began. Fort Wayne Public Library became Public Library of Fort Wayne and Allen County.
1944 – Main Library expansion began with purchase of Hollywood Building on Washington Boulevard to house administrative offices.
1952 – Young Adult Department opened in basement of main library, the first such department in the country.
1968 – building at Wayne and Webster Streets was dedicated August 21, 1968.
1977 – Construction of the new addition to the Main Library began.
1980 – Public Library of Fort Wayne and Allen County officially became Allen County Public Library by act of the state legislature on January 1.
1985 – Estimated 5,500 people ate 2,731 pizzas during system's 90th birthday celebration.
1997 – Library Board and staff began system-wide space needs analysis for all library facilities.
2001 – Allen County taxpayers approved bond financing of the $84 million library expansion project.
2007 – Renovation and expansion of main library nears completion. Grand opening held on January 27, 2007, with Randall T. Shepard, Chief Justice of the Indiana Supreme Court, among the speakers.
Genealogy.
The Fred J. Reynolds Historical Genealogy Department, located in the Main Library Branch, is the largest public genealogy department in North America, home to more than 350,000 printed volumes and 513,000 items of microfilm and microfiche. Only the Family History Library in Salt Lake City, a private institution, is larger. Allen County Public Library is also a partner of WeRelate, a collaborative online genealogy database currently providing access to over one million records and two million person pages. The website also provides the largest documented place wiki available.
Branches.
Ten of the Allen County Public Library's branches are located within Fort Wayne. Four branches are located in municipalities within Allen County (the name of the communities these four branches are located in is indicated next to each library's name).
Facilities.
The Main Library has five public meeting rooms that accommodate from six to 500 people. These rooms are available for use by the community for meetings and events. In addition, the building has six small group study rooms, a computer training room, and a theater which seats 230. Library branches also have meeting rooms and study rooms. The Main Library and the Georgetown branch have Maker Labs.
Access Fort Wayne.
Access Fort Wayne (AFW) is a Public-access television cable TV station which provides a variety of unique services for the library. AFW originates three Public-access television channels from the Main Library. Access 1 is channel 25 on Verizon and 55 on Comcast. Access 2 is channel 27 on Verizon and 57 on Comcast. Government-access television (GATV), City TV is channel 28 on Verizon and 58 on Comcast. The channels can only be viewed on Comcast and Verizon inside of Allen County. AFW is a full production facility with two TV studios and editing facilities.
Audio Reading Service.
Founded in 1979 as the Northeast Indiana Radio Reading Service (NEIRRS), the Audio Reading Service is a radio reading service for people who have visual, physical, learning, or language challenges to reading normal printed materials because of blindness, low-vision, a literacy issue, language barrier, or a physical condition. The service is for people with macular degeneration, diabetic retinopathy, arthritis, stroke, Parkinson's disease, and other conditions that affect reading traditional print. Under the leadership of library staff, volunteers read local news, grocery ads, cultural events, obituaries and other features from area newspapers, plus nearly 80 magazines and other publications.
Previously transmitted on an SCA broadcast, the Audio Reading Service can be heard on the HD Radio station 89.1 FM HD3, broadcast from Fort Wayne. HD Radios are loaned out at no cost by the library. In addition, the service is available via podcast, live web streaming, the Sero mobile app, and TuneIn; as a secondary audio program (SAP-French) on PBS 39.4; and system-wide at select area healthcare facilities.
The Audio Reading Service broadcasts 24 hours a day, seven days a week, and reaches an estimated 80,000 individuals. It became a part of the Allen County Public Library in 2004 and changed its name to Audio Reading Service in 2015. It is a member of the International Association of Audio Information Services (IAAIS).
WELT 95.7 FM.
WELT-LP is a non-commercial radio station, produced and delivered by members of the Fort Wayne community. From the studios at the downtown branch of the Allen County Public Library, WELT-LP's volunteers and staff promote programs with news about politics and community issues, plus music and entertainment performed by local, regional and national artists in Fort Wayne venues.


AM-0902

AM-0902 is a drug which acts as a potent and selective antagonist for the TRPA1 receptor, and has analgesic and antiinflammatory effects.