Ethanolamine-phosphate cytidylyltransferase

In enzymology, an ethanolamine-phosphate cytidylyltransferase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are CTP and ethanolamine phosphate, whereas its two products are diphosphate and CDP-ethanolamine.
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:ethanolamine-phosphate cytidylyltransferase. Other names in common use include phosphorylethanolamine transferase, ET, CTP-phosphoethanolamine cytidylyltransferase, phosphoethanolamine cytidylyltransferase, and ethanolamine phosphate cytidylyltransferase. This enzyme participates in aminophosphonate metabolism and glycerophospholipid metabolism.


Site-specific recombination

Site-specific recombination, also known as conservative site-specific recombination, is a type of genetic recombination in which DNA strand exchange takes place between segments possessing at least a certain degree of sequence homology. Enzymes known as site-specific recombinases (SSRs) perform rearrangements of DNA segments by recognizing and binding to short, specific DNA sequences (sites), at which they cleave the DNA backbone, exchange the two DNA helices involved, and rejoin the DNA strands. In some cases the presence of a recombinase enzyme and the recombination sites is sufficient for the reaction to proceed; in other systems a number of accessory proteins and/or accessory sites are required. Many different genome modification strategies, among these recombinase-mediated cassette exchange (RMCE), an advanced approach for the targeted introduction of transcription units into predetermined genomic loci, rely on SSRs.
Site-specific recombination systems are highly specific, fast, and efficient, even when faced with complex eukaryotic genomes. They are employed naturally in a variety of cellular processes, including bacterial genome replication, differentiation and pathogenesis, and movement of mobile genetic elements. For the same reasons, they present a potential basis for the development of genetic engineering tools.
Recombination sites are typically between 30 and 200 nucleotides in length and consist of two motifs with a partial inverted-repeat symmetry, to which the recombinase binds, and which flank a central crossover sequence at which the recombination takes place. The pairs of sites between which the recombination occurs are usually identical, but there are exceptions (e.g. attP and attB of λ integrase).
Classification: tyrosine- "vs." serine- recombinases.
Based on amino acid sequence homologies and mechanistic relatedness, most site-specific recombinases are grouped into one of two families: the tyrosine (Tyr) recombinase family or serine (Ser) recombinase family. The names stem from the conserved nucleophilic amino acid residue present in each class of recombinase which is used to attack the DNA and which becomes covalently linked to it during strand exchange. The earliest identified members of the serine recombinase family were known as resolvases or DNA invertases, while the founding member of the tyrosine recombinases, lambda phage integrase (using attP/B recognition sites), differs from the now well-known enzymes such as Cre (from the P1 phage) and FLP (from the yeast "Saccharomyces cerevisiae"). Famous serine recombinases include enzymes such as gamma-delta resolvase (from the Tn"1000" transposon), Tn3 resolvase (from the Tn3 transposon), and "φ"C31 integrase (from the "φ"C31 phage).
Although the individual members of the two recombinase families can perform reactions with the same practical outcomes, the families are unrelated to each other, having different protein structures and reaction mechanisms. Unlike tyrosine recombinases, serine recombinases are highly modular, as was first hinted by biochemical studies and later shown by crystallographic structures. Knowledge of these protein structures could prove useful when attempting to re-engineer recombinase proteins as tools for genetic manipulation.
Mechanism.
Recombination between two DNA sites begins by the recognition and binding of these sites – one site on each of two separate double-stranded DNA molecules, or at least two distant segments of the same molecule – by the recombinase enzyme. This is followed by synapsis, i.e. bringing the sites together to form the synaptic complex. It is within this synaptic complex that the strand exchange takes place, as the DNA is cleaved and rejoined by controlled transesterification reactions. During strand exchange, each double-stranded DNA molecule is cut at a fixed point within the crossover region of the recognition site, releasing a deoxyribose hydroxyl group, while the recombinase enzyme forms a transient covalent bond to a DNA backbone phosphate. This phosphodiester bond between the hydroxyl group of the nucleophilic serine or tyrosine residue conserves the energy that was expended in cleaving the DNA. Energy stored in this bond is subsequently used for the rejoining of the DNA to the corresponding deoxyribose hydroxyl group on the other DNA molecule. The entire reaction therefore proceeds without the need for external energy-rich cofactors such as ATP.
Although the basic chemical reaction is the same for both tyrosine and serine recombinases, there are some differences between them. Tyrosine recombinases, such as Cre or FLP, cleave one DNA strand at a time at points that are staggered by 6–8bp, linking the 3’ end of the strand to the hydroxyl group of the tyrosine nucleophile (Fig. 1). Strand exchange then proceeds via a crossed strand intermediate analogous to the Holliday junction in which only one pair of strands has been exchanged.
The mechanism and control of serine recombinases is much less well understood. This group of enzymes was only discovered in the mid-1990s and is still relatively small. The now classical members gamma-delta and Tn3 resolvase, but also new additions like φC31-, Bxb1-, and R4 integrases, cut all four DNA strands simultaneously at points that are staggered by 2 bp (Fig. 2). During cleavage, a protein–DNA bond is formed via a transesterification reaction, in which a phosphodiester bond is replaced by a phosphoserine bond between a 5’ phosphate at the cleavage site and the hydroxyl group of the conserved serine residue (S10 in resolvase).
It is still not entirely clear how the strand exchange occurs after the DNA has been cleaved. However, it has been shown that the strands are exchanged while covalently linked to the protein, with a resulting net rotation of 180°. The most quoted (but not the only) model accounting for these facts is the "subunit rotation model" (Fig. 2). Independent of the model, DNA duplexes are situated outside of the protein complex, and large movement of the protein is needed to achieve the strand exchange. In this case the recombination sites are slightly asymmetric, which allows the enzyme to tell apart the left and right ends of the site. When generating products, left ends are always joined to the right ends of their partner sites, and vice versa. This causes different recombination hybrid sites to be reconstituted in the recombination products. Joining of left ends to left or right to right is avoided due to the asymmetric “overlap” sequence between the staggered points of top and bottom strand exchange, which is in stark contrast to the mechanism employed by tyrosine recombinases.
The reaction catalysed by Cre-recombinase, for instance, may lead to excision of the DNA segment flanked by the two sites (Fig. 3A), but may also lead to integration or inversion of the orientation of the flanked DNA segment (Fig. 3B). What the outcome of the reaction will be is dictated mainly by the relative locations and orientations of the sites that are to be recombined, but also by the innate specificity of the site-specific system in question. Excisions and inversions occur if the recombination takes place between two sites that are found on the same molecule (intramolecular recombination), and if the sites are in the same (direct repeat) or in an opposite orientation (inverted repeat), respectively. Insertions, on the other hand, take place if the recombination occurs on sites that are situated on two different DNA molecules (intermolecular recombination), provided that at least one of these molecules is circular. Most site-specific systems are highly specialised, catalysing only one of these different types of reaction, and have evolved to ignore the sites that are in the "wrong" orientation.


Western blot normalization

Normalization of Western blot data is an analytical step that is performed to compare the relative abundance of a specific protein across the lanes of a blot or gel under diverse experimental treatments, or across tissues or developmental stages. The overall goal of normalization is to minimize effects arising from variations in experimental errors, such as inconsistent sample preparation, unequal sample loading across gel lanes, or uneven protein transfer, which can compromise the conclusions that can be obtained from Western blot data. Currently, there are two methods for normalizing Western blot data: (i) housekeeping protein normalization and (ii) total protein normalization.
Procedure.
Normalization occurs directly on either the gel or the blotting membrane. First, the stained gel or blot is imaged, a rectangle is drawn around the target protein in each lane, and the signal intensity inside the rectangle is measured. The signal intensity obtained can then be normalized with respect to the signal intensity of the loading internal control detected on the same gel or blot. When using protein stains, the membrane may be incubated with the chosen stain before or after immunodetection, depending on the type of stain.
Housekeeping protein controls.
Housekeeping genes and proteins, including β-Actin, GAPDH, HPRT1, and RPLP1, are often used as internal controls in western blots because they are thought to be expressed constitutively, at the same levels, across experiments. However, recent studies have shown that expression of housekeeping proteins (HKPs) can change across different cell types and biological conditions. Therefore, scientific publishers and funding agencies now require that normalization controls be previously validated for each experiment to ensure reproducibility and accuracy of the results.
Fluorescent antibodies.
When using fluorescent antibodies to image proteins in western blots, normalization requires that the user define the upper and lower limits of quantitation and characterize the linear relationship between signal intensity and the sample mass volume for each antigen. Both the target protein and the normalization control need to fluoresce within the dynamic range of detection. Many HKPs are expressed at high levels and are preferred for use with highly-expressed target proteins. Lower expressing proteins are difficult to detect on the same blot.
Fluorescent antibodies are commercially available, and fully characterized antibodies are recommended to ensure consistency of results.
When fluorescent detection is not utilized, the loading control protein and the protein of interest must differ considerably in molecular weight so they are adequately separated by gel electrophoresis for accurate analysis.
Membrane stripping.
Membranes need to be stripped and re-probed using a new set of detection antibodies when detecting multiple protein targets on the same blot. Ineffective stripping could result in a weak signal from the target protein. To prevent loss of the antigen, only three stripping incubations are recommended per membrane. It could be difficult to completely eliminate signal from highly-abundant proteins, so it is recommended that one detects lowly-expressed proteins first.
Exogenous spike-in controls.
Since HKP levels can be inconsistent between tissues, scientists can control for the protein of interest by spiking in a pure, exogenous protein of a known concentration within the linear range of the antibody. Compared to HKP, a wider variety of proteins are available for spike-in controls.
Total protein normalization.
In total protein normalization (TPN), the abundance of the target protein is normalized to the total amount of protein in each lane. Because TPN is not dependent on a single loading control, validation of controls and stripping/reprobing of blots for detection of HKPs is not necessary. This can improve precision (down to 0.1 µg of total protein per lane), cost-effectiveness, and data reliability.
Fluorescent stains and stain-free gels require special equipment to visualize the proteins on the gel/blot. Stains may not cover the blot evenly; more stain might collect towards the edges of the blot than in the center. Non-uniformity in the image can result in inaccurate normalization.
Pre-antibody stains.
Anionic dyes such as Ponceau S and Coomassie Brilliant Blue, and fluorescent dyes like Sypro Ruby and Deep Purple, are used before antibodies are added because they do not affect downstream imumunodetection.
Ponceau S is a negatively charged reversible dye that stains proteins a reddish pink color and is removed easily by washing in water. The intensity of Ponceau S staining decreases quickly over time, so documentation should be conducted rapidly. A linear range of up to 140 µg is reported for Ponceau S with poor reproducibility due to its highly time-dependent staining intensity and low signal-to-noise ratio.
Fluorescent dyes like Sypro Ruby have a broad linear range and are more sensitive than anionic dyes. They are permanent, photostable stains that can be visualized with a standard UV or blue-light transilluminator or a laser scan. Membranes can then be documented either on film or digitally using a CCD camera. Sypro Ruby blot staining is time-intensive and tends to saturate above 50 μg of protein per lane.
Post-antibody stains.
Amido black is a commonly used permanent post-antibody anionic stain that is more sensitive than Ponceau S. This stain is applied after imumunodetection.
Stain-free technology.
Stain-free technology employs an in-gel chemistry for imaging. This chemical reaction does not affect protein transfer or downstream antibody binding. Also, it does not involve staining/destaining steps, and the intensity of the bands remain constant over time.
Stain-free technology cannot detect proteins that do not contain tryptophan residues. A minimum of two tryptophans is needed to enable detection. The linear range for stain-free normalization is up to 80 µg of protein per lane for 18-well and up to 100 µg per lane for 12-well Criterion mid-sized gels. This range is compatible with typical protein loads in quantitative western blots and enables loading control calculations over a wide protein-loading range. A more efficient stain-free method has also recently become available. When using high protein loads, stain-free technology has demonstrated greater success than stains.


Trepipam

Trepipam (INN) is a dopamine receptor agonist of the benzazepine group that was never marketed.


Charles Joseph of Lorraine

Charles Joseph John Anthony Ignace Felix of Lorraine (), also known as "Charles III" in his capacity as the bishop of Olomouc (24 November 1680 – 4 December 1715), was a German prelate.
Born in Vienna, he was the second son of Charles V, Duke of Lorraine. He was bishop of Olomouc (1695–1711) and Prince-Bishop of Osnabrück (1698–1715), for which he was the successful candidate of the House of Palatinate, opposed by Brandenburg and, following some reverses and to the accompaniment of an enormous payment to the chapter of Trier, Charles Joseph was made archbishop and prince-elector of Trier (1711–1715), a political position of notable importance in the Holy Roman Empire. Already in 1711, he was able to make use of his electoral rights in the election of Charles VI, Holy Roman Emperor. He participated in the negotiations surrounding the end of the War of the Spanish Succession and succeeded in having the French occupying forces leave the Archbishopric in 1714. Charles Joseph died of smallpox during a visit in Vienna.


Jack Throck Watson

Jack Throck Watson (May 2, 1939 – September 3, 2016) was an American biochemist who was a professor of biochemistry and chemistry at the Michigan State University (MSU), where he was also Director of the MSU Mass Spectrometry Facility. While at MIT, Watson developed a gas chromatography–mass spectrometry interface, known as the Watson–Biemann separator, that removes helium from the gas chromatograph column effluent, thereby allowing analysis of less volatile and more polar compounds. Watson later worked on methods for the structure elucidation of peptides and proteins using fast atom bombardment and matrix-assisted laser desorption ionization (MALDI) mass spectrometry. After retirement in 2006, he continued to work on his introductory mass spectrometry textbook and teach short-courses in mass spectrometry.
Early life and education.
Jack Watson was born in was born May 2, 1939 in Casey, Iowa, to Jesse H. and Anne Watson. Jack grew up in a town of about 1,000 residents in northern Iowa, Nora Springs. His father was the area's school superintendent and he had one brother. After graduating from Nora Springs High School 1957, he went to Iowa State University, majoring in Chemistry and being a part of the University’s Air Force ROTC program for four years which accounts for the four years he spent on active duty in California and Texas. Before serving his Air Force obligation, after graduation Iowa State with a degree in Chemical Technology in 1961, he went to graduate school at the Massachusetts Institute Technology (MIT).
At MIT, Watson was a PhD candidate in the laboratory of Klaus Biemann, one of the most notable experts in organic mass spectrometry at the time. As soon as he graduated from MIT, Watson reported for duty in the United States Air Force in the San Francisco Bay area. A friend of his from high school, introduced Watson to Judith Sjoberg. Not long after that, they were married and moved to Brooks Air Base in San Antonio, Texas.
After completing his tour of duty in the Air Force, Watson took a one-year postdoctoral position in Strasbourg France at the Institut de Chimie, Université de Strasbourg under the direction of Robert Wolf. During this time and through the licensing of the Watson-Biemann gas separator to Thomson-CSF, for use in a gas chromatograph-mass spectrometer they ware manufacturing at the time, Watson made everlasting ties to the French Mass Spectrometry community.
Career.
After completing his postdoctoral fellowship in France, in 1969, Watson returned the United States and held a position as an Assistant Professor in the Department of Pharmacology at Vanderbilt University, in Nashville Tennessee. Jack was promoted to Associate Professor with tenure in 1974. While at Vanderbilt, Watson published the first edition of "Introduction to Mass Spectrometry: Biomedical, Environmental, and Forensic Applications" in 1976. It was the first book to include journal titles as part of the cited literature.
Harold G. (Harry) Walsh had just joined the ACS as director of the Short Course program. Walsh approached Watson and asked him to teach a course. Walsh also asked that Watson select someone from the mass spectrometry industry to co-teach the course. Watson had met O. David Sparkman, an American working for the French Gas Chromatography/Mass Spectrometry company, Riber, in Paris, a few months earlier. Watson asked Sparkman to contribute to the data systems part of the course. They taught the first session at the annual Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy in the Spring of 1978. They taught the course two more times that year at the annual ACS meetings and continued teaching into the first decade of the next millennium.
In 1980 Watson accepted a joint appointment in the Departments of Biochemistry and Chemistry at Michigan State University, East Lansing Michigan. He also became the director (Principal Investigator) of the National Institutes of Health (NIH) P41 Regional Resource in Mass Spectrometry at MSU. He remained director of the NIH facility until the NIH no longer funded such facilities and retired from his teaching position in 2006. The MS Facility continued to operate after funding stopped through the efforts of Watson, the Biochemistry and Chemistry Departments.
Personal life.
A friend of his from high school introduced Watson to Judith Sjoberg. Not long after that, they were married and moved to Brooks Air Base in San Antonio, Texas. They had two children in Nashville, Jennifer, born in 1970, and Brent born in 1972.
Legacy.
A Fellowship in Watson's name has been established at Michigan State University where recipients will be graduate students in the Department of Biochemistry and Molecular Biology.This is the “Jack Throck Watson Graduate Fellowship in Biochemistry Endowment.”


Cerner

Cerner Corporation is an American supplier of health information technology (HIT) services, devices, and hardware. As of February 2018, its products were in use at more than 27,000 facilities around the world. The company had more than 28,000 employees globally, with over 13,000 in Kansas City, Missouri. Its headquarters are in the suburb of North Kansas City, Missouri.
History.
Cerner was founded in 1979 by Neal Patterson, Paul Gorup, and Cliff Illig, who were colleagues at Arthur Andersen. Its original name was PGI & Associates but was renamed Cerner in 1984 when it rolled out its first system, PathNet. It went public in 1986. Cerner's client base grew steadily in the late 1980s, reaching 70 sites in 1987, 120 sites in 1988, 170 sites in 1989, and reaching 250 sites in 1990. Installations were primarily of PathNet systems.
During this time, Cerner was developing components of a Health Network Architecture (HNA), an integrated IT system designed to automate health care processes. Clients could purchase individual components or the whole system at one time. By 1994, more than 30 clients had purchased the full HNA system, while 100 clients had purchased multiple components of the system.
In 1997, the company introduced Cerner Millennium, an upgrade to its HNA system which incorporated all of the company's software offerings into one unified architecture. The introduction of Millennium contributed to significant growth for the company, with revenue increasing to $1.1 billion in 2005 from $245.1 million in 1997.
Cerner acquired IMC Health Care, Inc. in early 2010 to continue expanding its wellness services to outside commercial employers, pharmacies, and wellness programs.
In July 2010, president Trace Devanny left the company and Patterson became the company's president, in addition to his roles as chairman and chief executive officer. In September 2013, Zane Burke was named president, assuming the title from Patterson.
On August 5, 2014, Cerner announced its intent to purchase Siemens Health Services, the health information technology business of Germany's Siemens AG, for $1.3 billion. The acquisition was completed on February 2, 2015.
On July 29, 2015, Leidos Partnership for Defense Health, which includes Cerner, Accenture, and Leidos, was awarded a 10-year, $4.3 billion contract to overhaul and manage the electronic health records for the Department of Defense.
CEO and co-founder Neal Patterson died July 9, 2017.
On January 10, 2018, Brent Shafer was named Chairman and CEO and took over leadership responsibilities in February 2018.
In July 2019, Cerner announced that it would partner with Amazon Web Services.
On September 4, 2019, Cerner laid off 255 employees as part of an ongoing cost-cutting effort and reorganization.
In February 2020, Cerner sold parts of its healthcare IT business in Germany and Spain to German company CompuGroup Medical SE for EUR 225 million (USD 247.5M).
In 2020, Cerner announced the hire of three new C-suite executives including Jerome Labat as chief technology officer (CTO), Darrell Johnson as chief marketing officer (CMO) and William Mintz as chief strategy officer (CSO).
Products.
The company's products include:
The Cerner CCL (Cerner Command Language), is a SQL-like programming language developed by Cerner.
Controversy.
In 2001, a memo authored by CEO Patterson and sent to about 400 managers was leaked online. The memo, written in harsh language, was meant to motivate the managers to get more productivity out of employees and promised layoffs, a hiring freeze, closing of an "Associate Center," and the implementation of a punch-card system if Patterson did not see evidence of changes. Patterson's metric was the fullness of the company's Kansas City office lot at the hours of 8 a.m. and 5 p.m. The memo was widely seen as inflammatory and poor management, and Cerner's stock price fell 22% over three days.
In 2005, Cerner and other companies paid for a report by the RAND Corporation which predicted great efficiencies from electronic health records, including savings of $81 billion a year or more, which RAND now says is overstated. This report helped drive growth in the electronic health record and billions of dollars in federal incentives to hospitals and doctors. Cerner's revenue tripled from $1 billion in 2005 to a projected $3 billion in 2013. The study was criticized by the Congressional Budget Office for overstating potential savings. A 2013 reassessment of the 2005 report by the RAND Corporation said that the conversion had failed to produce savings and had mixed results in efficiency and patient care.
Canada.
In 2016, the Nanaimo BC hospital ER began using the system. Physicians and nurses have called it a "huge failure" with it increasing some types of errors and slowing down the emergency department. The system cost CAN$ 230 million dollars.
United Kingdom.
In 2014, a Coroner ruled that a three-year-old heart patient died as a result of a delay to his treatment at Bristol Royal Hospital for Children. The coroner ruled that the newly implemented Cerner Millennium booking system was responsible for the child not being seen or receiving treatment.
United States.
In 2002, the installation of a computerized health system by Cerner in the UPMC Children's Hospital of Pittsburgh made it harder for the doctors and nurses to do their jobs in emergency situations and resulted in a "disaster," according to Phillip Longman, a senior fellow at the New America Foundation. Longman wrote, "According to a study conducted by the hospital and published in the journal "Pediatrics", mortality rates for one vulnerable patient population—those brought by emergency transport from other facilities—more than doubled, from 2.8 percent before the installation to almost 6.6 percent afterward." Defenders of Cerner in the study charged that the Pittsburgh hospital did not adequately prepare for the transition to the Computerized Provider Order Entry (CPOE) system because it had simultaneously modified its pharmacy process, did not provide adequate wireless bandwidth, and did not have order sets pre-programmed on day one. They stated that other hospitals that more carefully planned the implementation did not experience the same problems.
In 2010, Girard Medical Center, Crawford County, Kansas, hired Cerner to install an electronic records system. But after receiving $1.3 million, Cerner employees failed to get the system running in time to qualify for federal incentive payments, and in September 2011, notified the hospital that it was abandoning the project, according to a lawsuit Girard filed against Cerner. Cerner and executives at Girard agreed that Girard did not have adequate staff to manage the acquisition and implementation of the system. As of June 2014, the case remained in arbitration. The outcome of the Girard case will likely be kept confidential due to contract provisions.
In 2012, Trinity Health, a large hospital in North Dakota, sued Cerner, claiming that Cerner's patient accounting solution didn't work correctly; the parties settled for $106M in 2014.
In 2014, a grand jury in California found that Cerner knew that the Ventura County healthcare agency was unprepared to complete a $32 million installation. Also in 2014, a $31 million Cerner implementation at the Athens Regional Health System in Georgia had many problems, leading to forced resignations by the CEO and the CIO.
Locations.
Cerner's world headquarters campus is at 2800 Rockcreek Parkway, North Kansas City, Missouri. It acquired additional space in Kansas City, Missouri in 2005 and in 2006 it acquired another location in Kansas City. In 2013, Cerner announced plans to redevelop 236 acres in south Kansas City, Missouri into an office park. The site was previously occupied by Bannister Mall, which was demolished in 2009. Cerner broke ground on the new campus on November 11, 2014. The $4.45 billion project intends to employ 16,000 new Cerner workers within the decade.
Cerner has offices in about 25 countries worldwide.


Nature Cell Biology

Nature Cell Biology is a highly selective peer-reviewed scientific journal covering research in cell biology published by Nature Research. It was established on 1999. The founding editor was Annette Thomas. The current editor-in-chief is Alexia-Ileana Zaromytidou.
According to the "Journal Citation Reports", the journal had a 2019 impact factor of 20.042.


MinE

The MinE protein is one of three proteins of the Min system encoded by the "minB" operon required to generate pole to pole oscillations prior to bacterial cell division as a means of specifying the midzone of the cell, as seen in E.coli.
History.
MinE was initially thought to assemble as a static ring at the cell center, thereby preventing the MinCD inhibition complex from localizing and binding there and instead influencing the complex to occupy each bacterial pole.
Raskin and de Boer later revealed a dynamic interaction of the Min proteins, where the unstable interaction between the proteins resulted in pole to pole oscillations, resulting in lower concentrations of MinCD complex at the cell center.
Function.
MinE is a topological specificity factor that counters the activity of MinCD division inhibitor at the mid-cell division site. MinE functions as a dimer and will bind to membrane-bound MinCD complex, altering its binding dynamics. MinE has membrane binding activity and can also interact with MinD only when on the membrane, suggesting a conformational change in MinD upon interacting with phospholipids that makes it vulnerable to MinE activity. MinE both positively and negatively regulates MinD interaction with membrane.
Deletion analysis has revealed that there exist two domains of interest in MinE, each with separate function. The N-Terminal domain contains the anti-MinCD domain that is necessary and sufficient
 to interact with MinD and counteract the MinCD-mediated division inhibition and to stimulate the ATPase activity of MinD, resulting in MinD’s detachment from the membrane after ATP hydrolysis. MinE is also known to form a ring near each pole. The purpose of this ring structure is to catalyze the release of membrane-bound MinD in turn, imparting regional specificity of Min protein localization.
Structure.
Nuclear Magnetic Resonance (NMR) spectroscopy of truncated MinE revealed a long alpha-helix as well as two anti-parallel beta-strands. These structures are hypothesized to mediate the homodimerization by interaction of alpha/beta structures. These structural studies also support the theory that the MinE ring structure may actually form a multimeric filamentous structure as a function of the interaction of these alpha and beta units. It has also been hypothesized that MinE may potentially form higher order structures, and may not simply be limited to ring formation.
Additional structural study and molecular imaging is needed to elucidate the higher order polymerization structures of MinE and to determine their dynamics on the remaining Min system. Recently, computational models have been applied to explore the limits of this system.


Lifestyle Pets

Lifestyle Pets, formerly Allerca, was a Delaware-based company which in the mid-2000s claimed to have bred hypoallergenic cats, although it was later revealed that the company's founder had previously engaged in multiple fraudulent enterprises.
According to company literature, through selective breeding the company achieved Ashera cats with significantly reduced levels of a particular protein that humans suffer allergic reactions to. The journal "Nature" reported in September 2006 that in an Allerca-funded study, Sheldon Spector, a clinical allergy expert at the University of California, Los Angeles, found the company's cats to be less allergy-inducing than the control. However, neither the study nor its underlying data have been published, and Spector himself advises caution as regards his study's interpretation as he used an experimental setup that is generally regarded as less than reliable.
A July 2006 article in the "San Diego Union-Tribune" reported that no details about the cats have been released, nor have there been any peer-reviewed studies published about the company's work. Cat experts also questioned whether the company's breeding model - stated at the time to be up to 10,000 cats by 2009 - was viable. Time named the company's cats among its list of Best Inventions for 2006. Allerca refuses to provide its kittens to clients whose allergy test results "indicate a level that includes HIGH". A cat based on the Domestic shorthair cost US$6,950 while ones based on the Siamese and Ashera cost $16,900 and $26,950 respectively.
In February 2006, Allerca was evicted from its San Diego headquarters, an address that doubled as the founder Simon Brodie's home. Brodie's previous businesses have "left behind unhappy clients, unpaid employees, debts, lawsuits, court judgments and liens..." The company Transgenic Pets sued Allerca and Brodie for theft of trade secrets and business plans. Allerca settled. The settlement agreed that Allerca would not re-enter the genetically engineered allergen free cat market until May 31, 2006.
For a while, Allerca offered a franchising program, possibly in violation of California law. Reports from Allerca indicate that they accepted at least partial payment from potential franchisees.
The company announced that on January 1, 2010 they will cease their breeding activities. However, their website remains active, and recent online complaints suggest that they are still in business. A report by ABC news in July 2013 maintains that the cats sold by Lifestyle Pets are no more hypoallergenic than any other cat.
Fel d 1.
Most human cat allergies are caused by Fel d 1. Allerca (and earlier, Geneticas) scientists tried to delete or disable the gene. The company now says it has "discovered" a breed of cats that had a mutant version of the protein that did not induce an allergic response. Since a number of Fel d 1 alleles are known and documented whereas it is presently impossible to deduce allergenicity from their DNA or protein sequence alone, the alleged mutant cats could exist, but their hypoallergenicity has not been proven.
Secrecy and lack of peer review.
Lifestyle Pets never submitted their scientific claims for peer review. In addition, the company was quite secretive. In a 2006 interview with the "San Diego Union-Tribune", the CEO at the time would not say where the company was located, how it was funded, how many people it employed, how many cats it had produced, or even where the cats were housed.
Fraudulent background of founder.
In 1994, Simon Brodie was convicted of seven counts of false accounting in the UK and sentenced to approximately two years in prison in connection with the collapse of a hot-air balloon company named Cloudhoppers. The "San Diego Union-Tribune" reported that Brodie planned to sell franchises at £200,000 each, but instead defrauded consumers and ran up hundreds of thousands of pounds in debts.
In 1999, Brodie incorporate Cerentis LLC in Delaware, United States, and the company sold software training packages. Peter Wood, 40, of Essex, England, purchased one such package for more than $50,000 with the understanding that he would receive training and then an employment offer. He did receive training at a California company named Alphalogix, but Brodie disappeared before the three-week course was completed. According to Los Angeles County court records, Brodie also failed to pay Alphalogix $30,600 in training fees. Mr. Wood said, "He's an extremely accomplished con man. He's obviously very good at what we call smoke and mirrors. He portrays a real professional company. But there's no depth to it at all."
Other Brodie-affiliated companies have included Integra Associates, Samba Wireless, Geneticas Life Sciences, ForeverPet, Genetiate, GeneSentinel, and Cyntegra.
Some customers did not receive any cats.
Discussions on forums and blogs continue to debate the status of this company and whether it is "vaporware". This has given rise to at least one web site complaining that creature has not been provided as promised. A report page has been opened at the Better Business Bureau of Wilmington, Delaware. It shows the current rating as "Unsatisfactory." Reports of customers receiving their Allerca GD kitten are sporadic, but they can be found on the web.
In September a man filed a law suit against Allerca for return of $7900 he paid for a hypoallergenic cat. Delivery was initially promised in eight months, but after repeated delays of the promised delivery date through 2008 and 2009, and a statement in March 2009 that the delivery scheduled had been affected by "decreased kitten production," the customer requested return of his money.
Another customer who did not receive a cat went to the news media from Live Oak, California, in 2009.
The company announced that they would stop taking orders for new kittens at the end of 2009 and instead focus on "veterinary diagnostic products," and that they would continue fulfilling previous orders through 2010 and 2011.
Ashera cats.
Lifestyle Pets also markets a hybrid cat under the name "Ashera." Several of the cats sold under the Ashera name were actually of the Savannah cat breed.


AFPep

AFPep (alpha fetoprotein peptide) is an orally-active, cyclic, 9-amino acid, peptide with a molecular weight of 969 Daltons and is derived from the anti-oncogenic active site (residues 472–479) of alpha fetoprotein (AFP). Using the standard amino acid abbreviations, AFPep has the sequence "cyclo"(EKTOVNOGN), where O is hydroxyproline. This peptide has been shown in experimental animal models to be efficacious in the prevention and treatment of ER+ breast cancer.
Biological activity.
Background.
Multiple births by a woman are strongly associated with a lower risk of developing breast cancer later in her life.
One of the contributing factors for this association appears to be the alpha fetoprotein (AFP) produced by the fetal liver, which crosses the placenta and enters into the maternal circulation. Pregnancy-associated protection from breast cancer is directly proportional to level of exposure to AFP. Furthermore, it has been demonstrated that tumor growth can be inhibited by AFP in animal models of breast cancer. It is speculated that AFP may induce apoptosis in pre-malignant breast tissue cells which would have later developed into malignancies.
Anti cancer effects.
Through mimicking the effects of AFP, AFPep inhibits the proliferation of estrogen receptor-positive human breast cancer cells growing in culture. It is also able to inhibit the estrogen stimulated growth of human breast cancer cells growing as xenografts in immunodeficient mice. According to a recent study, AFPep prevents the development of carcinogen-induced breast cancer in an animal model. Hence AFPep may have utility for preventing or treating estrogen receptor-positive breast cancer.
Mechanism of action.
AFPep inhibits estrogen-stimulated growth of immature mouse uterus and thus is antiestrogenic. In culture, AFPep inhibits the estrogen induced proliferation of T47D cells but has no effect on the basal growth. AFPep also inhibits phosphorylation of the estrogen receptor and activates the phosphorylation of p53.
AFPep has been shown to bind the heat shock protein Hsp72. Hsp72 together with Hsp90 form a heterocomplex with the estrogen receptor. Hence AFPep through interaction with Hsp72 controls the ligand binding and transcriptional activation of the estrogen receptor.
Combination therapy with tamoxifen.
AFPep increases the efficacy and decreases the toxicities of tamoxifen.
Tamoxifen has been a very effective drug for the treatment of estrogen receptor-positive breast cancer. But tamoxifen has certain toxicities and side effects such as uterine hyperplasia which can lead to endometrial cancer. Moreover, some breast cancers acquire resistance to tamoxifen during the course of treatment and few are totally resistant to it. It has been established that AFPep when used in combination with tamoxifen, reduces the uterine hyperplasia and increases the antitumour effects of tamoxifen. A rational combination of AFPep and tamoxifen may prove to be a better chemopreventive or chemotherapeutic approach against estrogen receptor-positive breast cancer.
Route of administration.
AFPep active whether administered intraperitoneally, subcutaneously or orally.


Lipid bilayer phase behavior

One property of a lipid bilayer is the relative mobility (fluidity) of the individual lipid molecules and how this mobility changes with temperature. This response is known as the phase behavior of the bilayer. Broadly, at a given temperature a lipid bilayer can exist in either a liquid or a solid phase. The solid phase is commonly referred to as a “gel” phase. All lipids have a characteristic temperature at which they undergo a transition (melt) from the gel to liquid phase. In both phases the lipid molecules are constrained to the two dimensional plane of the membrane, but in liquid phase bilayers the molecules diffuse freely within this plane. Thus, in a liquid bilayer a given lipid will rapidly exchange locations with its neighbor millions of times a second and will, through the process of a random walk, migrate over long distances.
Motion constraints.
In contrast to this large in-plane mobility, it is very difficult for lipid molecules to flip-flop from one side of the lipid bilayer to the other. In a phosphatidylcholine-based bilayer this process typically occurs over a timescale of weeks. This discrepancy can be understood in terms of the basic structure of the bilayer. For a lipid to flip from one leaflet to the other, its hydrated headgroup must cross the hydrophobic core of the bilayer, an energetically unfavorable process. Unlike liquid phase bilayers, the lipids in a gel phase bilayer are locked in place and exhibit neither flip-flop nor lateral mobility. Due to this limited mobility, gel bilayers lack an important property of liquid bilayers: the ability to reseal small holes. Liquid phase bilayers can spontaneously heal small voids, much the same way a film of oil on water could flow in to fill a gap. This functionality is one of the reasons that cell membranes are usually composed of fluid phase bilayers. Motion constraints on lipids in lipid bilayers are also imposed by presence of proteins in biological membranes, especially so in the annular lipid shell 'attached' to surface of integral membrane proteins.
Physical origins.
The phase behavior of lipid bilayers is largely determined by the strength of the attractive Van der Waals interactions between adjacent lipid molecules. The extent of this interaction is in turn governed by how long the lipid tails are and how well they can pack together. Longer tailed lipids have more area over which to interact, increasing the strength of this interaction and consequently decreasing the lipid mobility. Thus, at a given temperature, a short-tailed lipid will be more fluid than an otherwise identical long-tailed lipid. Another way of expressing this would be to say that the gel to liquid phase transition temperature increases with increasing number of carbons in the lipid alkane chains. Saturated phosphatidylcholine lipids with tails longer than 14 carbons are solid at room temperature, while those with fewer than 14 are liquid. This phenomenon is analogous to the fact that paraffin wax, which is composed of long alkanes, is solid at room temperature, while octane (gasoline), a short alkane, is liquid.
Aside from chain length, transition temperature can also be affected by the degree of unsaturation of the lipid tails. An unsaturated double bond can produce a kink in the alkane chain, disrupting the regular periodic structure. This disruption creates extra free space within the bilayer which allows additional flexibility in the adjacent chains. It is this disruption of packing that leads to lower transition temperatures with increasing double bonds. This is a particularly powerful effect; decreasing the overall chain length by one carbon usually alters the transition temperature of a lipid by ten degrees Celsius or less, but adding a single double bond can decrease the transition temperature by fifty degrees or more (see table). An example of this effect can be noted in everyday life as butter, which has a large percentage saturated fats, is solid at room temperature while vegetable oil, which is mostly unsaturated, is liquid.
Mixed systems.
Bilayers need not be composed of a single type of lipid and, in fact, most natural membranes are a complex mixture of different lipid molecules. Such mixtures often exhibit properties intermediate to their components, but are also capable of a phenomenon not seen in single component systems: phase separation. If some of the components are liquid at a given temperature while others are in the gel phase, the two phases can coexist in spatially separated populations. This phase separation plays a critical role in biochemical phenomena because membrane components such as proteins can partition into one or the other phase and thus be locally concentrated or activated.
Cholesterol.
The presence of cholesterol exerts a profound but complicated influence on lipid bilayer properties because of its unique physical characteristics. Although it is a lipid, cholesterol bears little resemblance to a phospholipid. The hydrophilic domain of cholesterol is quite small, consisting of a single hydroxyl group. Adjacent to this hydroxyl group is a rigid planar structure composed of several fused rings. At the opposite end of the ring structure is a short single chain tail. It has been known for decades that the addition of cholesterol to a fluid phase bilayer decreases its permeability to water. The mode of this interaction has more recently been shown to be due to cholesterol intercalating between lipid molecules, filling in free space and decreasing the flexibility of surrounding lipid chains. This interaction also increases the mechanical rigidity of fluid membrane lipid bilayers and decreases their lateral diffusion coefficient. In contrast, the addition of cholesterol to gel phase bilayers disrupts local packing order, increasing the diffusion coefficient and decreasing the elastic modulus. Interactions of cholesterol with multi-component systems are even more complicated, as these can result in intricate phase diagrams. One lipid-cholesterol system that has recently been studied intently is the lipid raft. Lipid rafts are cholesterol-enriched gel domains that have been potentially implicated in certain cell signaling processes, but the subject remains controversial, with some researchers doubting even their existence in vivo.
Lipid polymorphism.
Mixed lipid liposomes can undergo changes into different phase dispersion structures, called lipid polymorphisms, for example, spherical micelles, lipid bilayer lamellae and hexagonal phase cylinders, depending on physical and chemical changes in their microenvironment.
Phase transition temperature of liposomes and biological membranes can be measured using calorimetry, magnetic resonance spectroscopy and other techniques.


Kelba (genus)


Molybdenum cofactor guanylyltransferase

Molybdenum cofactor guanylyltransferase (, "MobA", "MoCo guanylyltransferase") is an enzyme with systematic name "GTP:molybdenum cofactor guanylyltransferase". This enzyme catalyses the following chemical reaction:
Catalyses the guanylation of the molybdenum cofactor.


Methanethiol oxidase

In enzymology, a methanethiol oxidase () is an enzyme that catalyzes the chemical reaction
The 3 substrates of this enzyme are methanethiol, O2, and H2O, whereas its 3 products are formaldehyde, hydrogen sulfide, and H2O2.
This enzyme belongs to the family of oxidoreductases, specifically those acting on a sulfur group of donors with oxygen as acceptor. The systematic name of this enzyme class is methanethiol:oxygen oxidoreductase. Other names in common use include methylmercaptan oxidase, methyl mercaptan oxidase, (MM)-oxidase, and MT-oxidase.


D-ribitol-5-phosphate cytidylyltransferase

In enzymology, a D-ribitol-5-phosphate cytidylyltransferase () is an enzyme that catalyzes the chemical reaction
Thus, the two substrates of this enzyme are CTP and D-ribitol 5-phosphate, whereas its two products are diphosphate and CDP-ribitol.
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:D-ribitol-5-phosphate cytidylyltransferase. Other names in common use include CDP ribitol pyrophosphorylase, cytidine diphosphate ribitol pyrophosphorylase, ribitol 5-phosphate cytidylyltransferase, and cytidine diphosphoribitol pyrophosphorylase. This enzyme participates in pentose and glucuronate interconversions.


Aravind L. Iyer

Aravind L. Iyer is an evolutionary biologist at the National Center for Biotechnology Information in Bethesda, Maryland, United States. Since 2002, he has been Principal Investigator of the Computational Biology Branch. In his research publications he goes by the name of L. Aravind. He was brought up for the most part in Pune. He obtained his master's degree in Biotechnology and subsequently moved to the Texas A&M University to eventually complete his PhD there in December 1999.


GRIK2

Glutamate ionotropic receptor kainate type subunit 2, also known as ionotropic glutamate receptor 6 or GluR6, is a protein that in humans is encoded by the "GRIK2" (or "GLUR6") gene.
Function.
This gene encodes a subunit of a kainate glutamate receptor. This receptor may have a role in synaptic plasticity, learning, and memory. It also may be involved in the transmission of visual information from the retina to the hypothalamus. The structure and function of the encoded protein is influenced by RNA editing. Alternatively spliced transcript variants encoding distinct isoforms have been described for this gene.
Clinical significance.
Homozygosity for a GRIK2 deletion-inversion mutation is associated with non-syndromic autosomal recessive mental retardation.
Interactions.
GRIK2 has been shown to interact with:
RNA Editing.
Pre-mRNA for several neurotransmitter receptors and ion channels are substrates for ADARs, including AMPA receptor subunits (GluR2, GluR3, GluR4) and kainate receptor subunits (GluR5, GluR6). Glutamate-gated ion channels are made up of four subunits per channel, with each subunit contributing to the pore loop structure. The pore loop structure is similar to that found in K+ channels (e.g. the human Kv1.1 channel, whose pre-mRNA is also subject to A to I RNA editing). The diversity of ionotropic glutamate receptor subunits, as well as RNA splicing, is determined by RNA editing events of the individual subunits, explaining their extremely high diversity.
Type.
The type of RNA editing that occurs in the pre-mRNA of GluR6 is Adenosine to Inosine (A to I) editing.
A to I RNA editing is catalyzed by a family of adenosine deaminases acting on RNA (ADARs) that specifically recognize adenosines within double-stranded regions of pre-mRNAs and deaminate them to inosine. Inosines are recognised as guanosine by the cell's translational machinery. There are three members of the ADAR family ADARs 1-3 with ADAR1 and ADAR2 being the only enzymatically-active members. ADAR1 and ADAR2 are widely expressed in tissues, while ADAR3 is restricted to the brain, where it is though tot have a regulatory role. The double-stranded regions of RNA are formed by base-pairing between residues close to region of the editing site, with residues usually in a neighboring intron, though they can occasionally be located in an exonic sequence. The region that forms base pairs with the editing region is known as an Editing Complementary Sequence (ECS).
ADARs bind interact directly with the dsRNA substrate via their double-stranded RNA binding domains. If an editing site occurs within a coding sequence, the result could be a codon change. This can lead to translation of a protein isoform due to a change in its primary protein structure. Therefore, editing can also alter protein function. A to I editing occurs in a noncoding RNA sequences such as introns, untranslated regions (UTRs), LINEs, and SINEs (especially Alu repeats). The function of A to I editing in these regions is thought to involve creation of splice sites and retention of RNAs in the nucleus amongst others.
Location.
The pre-mRNA of GLUR6 is edited at amino acid positions 567, 571, and 621.
The Q/R position, which gets its name as editing results in an codon change from a glutamine (Q) codon (CAG) to an arginine (R) codon (CGG), is located in the "pore loop" of the second membrane domain (M2). The Q/R site of GluR6 pre-mRNA occurs in an asymmetrical loop of three exonic and four intronic nucleotides. The Q/R editing site is also observed in GluR2 and GluR5. The Q/R site is located in a homologous position in GluR2 and in GluR6.
GluR-6 is also edited at I/V and Y/C sites, which are found in the first membrane domain (M1). At the I/V site, editing results in a codon change from (ATT) isoleucine (I) to (GTT) valine (V), while at the Y/C site, the codon change is from (TAC) tyrosine (Y) to (TGC) cysteine (C).
The RNAfold program characterised a putative double-stranded RNA (dsRNA) conformation around the Q/R site of the GluR-6 pre-mRNA. This sequence is necessary for editing at the site to occur. The possible editing complementary sequence was observed from transcript analysis to be 1.9 kb downstream from the editing site within intron 12.
The ECS for the editing sites in M1 has yet to be identified but it is likely to occur at a considerable distance from the editing sites.
Regulation.
Editing of the Q/R site in GluR6 pre-mRNA has been demonstrated to be developmentally regulated in rats, ranging from 0% in rat embryo to 80% at birth. This is different from the AMPA receptor subunit GluR2, which is nearly 100% edited and is not developmentally regulated.
Significant amounts of both edited and non-edited forms of GluR6 transcripts are found in the adult brain. The receptor is 90% edited in all grey matter structures, while in white matter, the receptor is edited in just 10% of cases.
Frequency increases from 0% in rat embryo to 85% in adult rat.
Consequences.
Structure.
The primary GluR6 transcripts can be edited in up to three positions. Editing at each of the three positions affects Ca2+ permeability of the channel.
Function.
Editing plays a role in the electrophysiology of the channel.
Editing at the Q/R site has been deemed to be nonessential in GluR6. It has been reported that the unedited version of GluR6 functions in the regulation of synaptic plasticity. The edited version is thought to inhibit synaptic plasticity and reduce seizure susceptibility.
Mice lacking the Q/R site exhibit increased long term potentiation and are more susceptible to kainate induced seizures. The number of seizures is inversely correlated with the amount of RNA editing. Human GluR6 pre-mRNA editing is increased during seizures, possibly as an adaptive mechanism.
Up to 8 different protein isoforms can occur as a result of different combinations of editing at the three sites, giving rise to receptor variants with differing kinetics. The effect of Q/R site editing on calcium permeability appears to be dependent on editing of the I/V and Y/C sites. When both sites in TM1 (I/V and Y/C) are edited, Q/R site editing is required for calcium permeability. On the contrary, when neither the I/V nor the Y/C site is edited, receptors demonstrate high calcium permeability regardless of Q/R site editing. The co-assembly of these two isoforms generate receptors with reduced calcium permeability.
RNA editing of the Q/R site can affect inhibition of the channel by membrane fatty acids such as arachidonic acid and docosahexaenoic acid For Kainate receptors with only edited isforms, these are strongly inhibited by these fatty acids, however inclusion of just one non-edited subunit is enough to abolish this effect.
Dysregulation.
Kainate-induced seizures in mice are used as a model of temporal lobe epilepsy in humans. Despite mice deficient in editing at the Q/R site of GluR6 showing increased seizure susceptibility, tissue analysis of human epilepsy patients did not show reduced editing at this site.


Kinetic Concepts

Kinetic Concepts, Inc., (KCI) is a global corporation that produces medical technology related to wounds and wound healing. KCI produced the first product developed specifically for negative pressure wound therapy. In 2013, the company employed 5,000 people and marketed its products in more than 25 countries. Its headquarters is in San Antonio, Texas.
KCI, LifeCell and Systagenix operated under the Acelity brand.
In October 2019, Acelity and its KCI subsidiaries worldwide were acquired by 3M for $6.7 billion, including assumption of debt and other adjustments.
Operations.
KCI is composed of three business units: Active Healing Solutions, LifeCell and Therapeutic Support Systems, that operate in the wound care, regenerative medicine and therapeutic support systems markets. The largest of these business units is dedicated to wound care primarily negative pressure wound therapy for the treatment of traumatic wounds, pressure ulcers, chronic wounds and diabetic ulcers but also assistance with surgery. The therapeutic support systems developed and supplied by KCI are largely for the treatment and prevention of complications associated with patient immobility. These include support surfaces for hospital beds and home patients designed to address pulmonary complications in immobile patients. KCI also develops specialty beds for use in hospitals or in long-term care facilities. KCI expanded into the regenerative medicine market in 2008. Through subsidiary company LifeCell Corporation, KCI develops and supplies tissue-based products. These include tissue-based treatments used in surgical procedures to repair soft-tissue.
Corporate history.
KCI was founded in 1976 in San Antonio, Texas, by James R. Leininger, then an emergency room physician who wanted to help prevent the pulmonary complications associated with immobility. Over time, the company developed or acquired a line of therapeutic specialty beds, introducing a specialty bed for acute care patients with pulmonary complications. Initially KCI's product development focused on therapeutic beds and surfaces then expanded to introduce the first commercial negative pressure wound therapy products in the mid-1990s. KCI acquired regenerative medicine company, LifeCell, in 2008 in a non-hostile transaction for 1.7 billion. In January 2012, LifeCell was made a sister company to KCI. In 2013, LifeCell and KCI recombined into one company.
Financial information.
From the founding of the company in 1976 to 1988, the revenue of KCI grew to 153.2 million. The growth in revenue allowed KCI to go public and trade on the New York Stock Exchange (NYSE) in 1988. KCI stock was traded on the NYSE until 1997, when the company went private. In February 2004, KCI became a publicly listed company for the second time and was listed on the NYSE under the ticker symbol KCI.
By the late 2000s, the company's revenue was over $1 billion and reported an increase of 17 percent in 2007 to $1.61 billion. Revenue for 2008 was $1.88 billion, increasing to $1.99 billion in 2009 and in 2010 increased to $2.02 billion. On November 4, 2011, the company went private again at a price of $68.50 per share.


5-DBFPV

5-DBFPV (also known as 5-dihydrobenzofuranpyrovalerone and 3-desoxy-MDPV) is a stimulant of the cathinone class that has been sold online as a designer drug. It is an analogue of MDPV where the methylenedioxyphenyl group has been replaced by dihydrobenzofuran.
Legal status.
5-DBFPV is illegal in Sweden as of 26. January 2016.


Annexin Pharmaceuticals

Annexin Pharmaceuticals is a Swedish privately held biotech company developing new therapeutic approaches for inflammatory cardiovascular diseases. The concept for the therapy is based on the anti-inflammatory properties of Annexin A5, a body own protective protein that acts simultaneously against several key pathogenic mechanisms of cardiovascular diseases. The company is currently focusing on treatment of peripheral artery disease (PAD).


Jean Fan

Jean Fan is an American biomedical engineer. She won a Nature Research Award for Inspiring Women.
Life.
She graduated from Johns Hopkins University, where she studied with Rachel Karchin, and Harvard University.
She teaches at The Innovation Institute. She was the founder of cuSTEMized. She teaches at Johns Hopkins University.


L-amino-acid dehydrogenase

In enzymology, a L-amino-acid dehydrogenase () is an enzyme that catalyzes the chemical reaction
The 3 substrates of this enzyme are L-amino acid, H2O, and NAD+, whereas its 4 products are 2-oxo acid, 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-amino-acid:NAD+ oxidoreductase (deaminating).


Reverse transfection

Reverse transfection is a technique for the transfer of genetic material into cells. As DNA is printed on a glass slide for the transfection process (the deliberate introduction of nucleic acids into cells) to occur before the addition of adherent cells, the order of addition of DNA and adherent cells is reverse that of conventional transfection. Hence, the word “reverse” is used.
Process.
Transfection-mix preparation for slide printing.
A DNA-gelatin mixture may be used for printing onto a slide. Gelatin powder is first dissolved in sterile Milli-Q water to form a 0.2% gelatin solution. Purified DNA plasmid is then mixed with the gelatin solution, and the final gelatin concentration is kept greater than 0.17%. Besides gelatin, atelocollagen and fibronetin are also successful transfection vectors for introducing foreign DNA into the cell nucleus.
Slide printing of DNA-gelatin mixture.
After the DNA-gelatin mixture preparation, the mixture is pipetted onto a slide surface and the slide is placed in a covered petri dish. A desiccant is added to the dish to dry up the solution. Finally, cultured cells are poured into the dish for plasmid uptake. However, with the invention of different types of microarray printing systems, hundreds of transfection mixes (containing different DNA of interest) may be printed on the same slide for cell uptake of plasmids. There are two major types of microarray printing systems manufactured by different companies: contact and non-contact printing systems.
An example of a non-contact printing system is the Piezorray Flexible Non-contact Microarraying System. It uses pressure control and a piezoelectric collar to squeeze out consistent drops of approximately 333 pL in volume. The PiezoTip dispensers do not contact the surface to which the sample is dispensed; thus, contamination potential is reduced and the risk of disrupting the target surface is eliminated. An example of a contact printing system is the SpotArray 72 (Perkin Elmer Life Sciences) contact-spotting system. Its printhead can accommodate up to 48 pins, and creates compact arrays by selectively raising and lowering subsets of pins during printing. After printing, the pins are washed with a powerful pressure-jet pin washer and vacuum-dried, eliminating carryover. Another example of a contact printing system is the Qarray system (Genetix). It has three types of printing systems: QArray Mini, QArray 2 and QArray Max. After printing, the solution is allowed to dry up and the DNA-gelatin is stuck tightly in position on the array.
HybriWell in reverse transfection.
First, adhesive from the HybriWell is peeled off and the HybriWell is attached over the area of the slide printed with the gelatin-DNA solution. Second, 200ul of transfection mix is pipetted into one of the HybriWell ports; the mixture will distribute evenly over the array. The array is then incubated, with temperature and time dependent on the cell types used. Third, the transfection mix is pipetted away and the HybriWell removed with a thin-tipped forceps. Fourth, the printed slide treated with transfection reagent is placed into a square dish with the printed side facing up. Fifth, the harvested cells are gently poured onto the slides (not on the printed areas). Finally, the dish is placed in a 37°C, 5% CO2 humidified incubator and incubated overnight.
Other reverse-transfection reagents.
Effectene Reagent is used in conjunction with the enhancer and the DNA condensation buffer (Buffer EC) to achieve high transfection efficiency. In the first step of Effectene–DNA complex formation, the DNA is condensed by interaction with the enhancer in a defined-buffer system. Effectene Reagent is then added to the condensed DNA to produce condensed Effectene–DNA complexes. The Effectene–DNA complexes are mixed with the medium and directly added to the cells.
Effectene Reagent spontaneously forms micelle structures exhibiting no size or batch variation (as may be found with pre-formulated liposome reagents). This feature ensures reproducibility of transfection-complex formation. The process of highly condensing DNA molecules and then coating them with Effectene Reagent is an effective way to transfer DNA into eukaryotic cells.
Advantages and disadvantages.
The advantages of reverse transfection (over conventional transfection) are:
The disadvantages of reverse transfection are:


ChIA-PET

Chromatin Interaction Analysis by Paired-End Tag Sequencing (ChIA-PET or ChIA-PETS) is a technique that incorporates chromatin immunoprecipitation (ChIP)-based enrichment, chromatin proximity ligation, Paired-End Tags, and High-throughput sequencing to determine "de novo" long-range chromatin interactions genome-wide.
Genes can be regulated by regions far from the promoter such as regulatory elements, insulators and boundary elements, and transcription-factor binding sites (TFBS). Uncovering the interplay between regulatory regions and gene coding regions is essential for understanding the mechanisms governing gene regulation in health and disease (Maston et al., 2006). ChIA-PET can be used to identify unique, functional chromatin interactions between distal and proximal regulatory transcription-factor binding sites and the promoters of the genes they interact with.
ChIA-PET can also be used to unravel the mechanisms of genome control during processes such as cell differentiation, proliferation, and development. By creating ChIA-PET interactome maps for DNA-binding regulatory proteins and promoter regions, we can better identify unique targets for therapeutic intervention (Fullwood & Yijun, 2009).
Methodology.
The ChIA-PET method combines ChIP-based methods (Kuo & Allis, 1999), and Chromosome conformation capture (3C), to extend the capabilities of both approaches. ChIP-Sequencing (ChIP-Seq) is a popular method used to identify TFBS while 3C has been used to identify long-range chromatin interactions (Dekker et al., 2002). However, both suffer from limitations when used independently to identify de-novo long-range interactions genome wide. While ChIP-Seq is typically used for genome-wide identification of TFBS (Barski et al., 2007; Wei et al., 2006), it provides only linear information of protein binding sites along the chromosomes (but not interactions between them), and can suffer from high genomic background noise (false positives).
While 3C is capable of analyzing long-range chromatin interactions, it cannot be used genome
wide and, like ChIP-Seq, also suffers from high levels of background noise. Since the noise increases in relation to the distance between interacting regions (max 100kb), laborious and tedious controls are required for accurate characterization of chromatin interactions (Dekker et al., 2006).
The ChIA-PET method successfully resolves the issues of non-specific interaction noise found in ChIP-Seq by sonicating the ChIP fragments in order to separate random attachments from specific interaction complexes. The next step, which is referred to as enrichment, reduces complexity for genome-wide analysis and adds specificity to chromatin interactions bound by pre-determined TFs (transcription factors).
The ability of 3C approaches to identify long-range interactions is based on the theory of proximity ligation. In regards to DNA inter-ligation, fragments that are tethered by common protein complexes have greater kinetic advantages under dilute conditions, than those freely diffusing in solution or anchored in different complexes. ChIA-PET takes advantage of this concept by incorporating linker sequences onto the free ends of the DNA fragments tethered to the protein complexes. In order to build connectivity of the fragments tethered by regulatory complexes, the linker sequences are ligated during nuclear proximity ligation.
Therefore, the products of linker-connected ligation can be analyzed by ultra-high-throughput PET sequencing and mapped to the reference genome. Since ChIA-PET is not dependent on specific sites for detection as 3C and 4C are, it allows unbiased, genome-wide de-novo detection of chromatin interactions (Fullwood et al., 2009).
Workflow.
Wet-lab portion of the workflow:
Dry-lab portion of the workflow:
PET extraction, mapping, and statistical analyses
The PET tags are extracted and mapped to the reference human genome "in silico".
Identification of ChIP enriched peaks (binding sites)
Self-ligated PET are used for identifying ChIP enriched sites because they provide the most reliable mapping (20 + 20 bit/s) to the reference genome.
ChIP enrichment peak-finding algorithm
A called peak is considered a binding site if there are multiple overlapping self-ligated PETs.
The false discovery rate (FDR) is determined using statistical simulations to estimate the random background of PET-derived virtual DNA overlaps, and the estimated background noise.
Filtering of repetitive DNA (affects non-specific binding)
Satellite regions and binding sites present in regions with severe structural variations are removed.
ChIP enrichment count
The numbers of self-ligation and inter-ligation PETs (within + 250 bp window) are reported at each site. The total number of self-ligated and inter-ligated PETs at a specific site is called the ChIP enrichment count.
Figure 6. PET Classification: Uniquely aligned PET sequences can be classified by whether they are derived from one DNA fragment or two DNA fragments.
If the two tags of a PET are mapped on the same chromosome with the genomic span in the range of ChIP DNA fragments (less than 3 Kb), with expected self-ligation orientation and on the same strand, they are considered to be derived from a self-ligation of a single ChIP DNA fragment, and considered a self-ligation PET.
If a PET does not fit into these criteria, then the PET most likely resulted from a ligation product between two DNA fragments and referred to as an inter-ligation PET. The two tags of an inter-ligation PETs do not have fixed tag orientations, might not be found on the same strands, might have any genomic span, and might not map to the same chromosome.
If the two tags of an inter-ligation PET are mapped in the same chromosome but with a span > 3 Kb in any orientation, then these PETs are called intrachromosomal inter-ligation PETs.
PETs which are mapped to different chromosomes are called interchromosomal inter-ligation PETs.
Figure 7. Proposed mechanism showing how distal regulatory elements can initiate long-range chromatin interactions involving promoter regions of target genes.
The interactions form DNA loop structures with multiple TFBS at the anchoring center. Small loops might package genes near the anchoring center in a tight sub-compartment, which could increase the local concentration of regulatory proteins for enhanced transcriptional activation. This mechanism might also enhance transcription efficiency, allowing RNA pol II to cycle the tight circular gene templates. The large interaction loops are more likely to link together distant genes at either end of the loop residing near anchor sites for coordinated regulation, or could separate genes in long loops to prevent their activation. Adapted from Fullwood et al. (2009).
Strengths and weaknesses.
Advantages of the ChIA-PET method
Weaknesses
History.
Fullwood et al. (2009), used ChIA-PET to detect and map the chromatin interaction network mediated by estrogen receptor alpha (ER-alpha) in human cancer cells. The resulting global chromatin interactome map revealed that remote ER-alpha-binding sites were also anchored to gene promoters through long-range chromatin interactions suggesting that ER-alpha functions by extensive chromatin looping in order to bring genes together for coordinated transcriptional regulation.
Analysis and software.
Software typically used in a ChIA-PET experiment
ELAND
Maps ChIP enriched DNA fragments to the reference human genome.
RepeatMasker
In-silico masking of repetitive elements.
Monte Carlo simulation
Used to estimate the false discovery rates.
PET-Tool
A software suite for processing and managing of Paired-End di-Tag sequence data.
ChIA-PET Tool
A software suite for processing ChIA-PET data.ChIA-PET Tool web site ChIA-PET Tool paper
Alternatives.
Chromatin immunoprecipitation:
ChIP-Seq, ChIP-PET, ChIP-SAGE, ChIP-CHIP.
Chromosome conformation capture:
2-C, 3-C, 4-C, 5-C.
Paired-end tags:
PET.


Signal patch

A protein signal patch contains information to send a given protein to the indicated location in the cell. It is made up of amino acid residues that are distant to one another in the primary sequence, but come close to each other in the tertiary structure of the folded protein (see red patch in the diagram). Signal patches, unlike some signal sequences, are not cleaved from the mature protein after sorting. They are very difficult to predict. Nuclear localization signals are often signal patches although signal sequences also exist. They are found on proteins destined for the nucleus and enable their selective transport from the cytosol into the nucleus through the nuclear pore complexes.


Christopher James Davis

Christopher James Davis (1842 – 27 November 1870) was a British-Barbadian physician. Educated in Europe, he volunteered to help sick and injured during the Franco-Prussian war in 1870, and died from smallpox.
Childhood.
Davis was born in Bridgetown, Barbados, in a family of ten children. His family were Wesleyan Methodists. His father was British, his mother a Barbadian. "Of all my children," wrote his mother, "he seemed the most tender and considerate for me, and would weep himself to sleep if anything tired me."
Career.
Davis studied to become a school teacher and was a lay-preacher in the Wesleyan Methodist Church. The Plymouth Brethren movement had recently spread from British Guyana to Barbados and other West Indian colonies. Their teachings were imbibed by Davis and he began to propagate them. Three of his own family also joined the Brethren.
Limerick-born Dr. Thomas Mackern of Blackheath, South East London, visited the West Indies in 1866 to preach the gospel and to promote the literature work of the Plymouth Brethren in the islands. He met numerous Christians who had then recently broken with their denominational associations. He wrote, "They had begun 'amidst much opposition from professing Christians' to gather 'simply to the name of the Lord Jesus' – which meant that Brethren assemblies were functioning. The one chiefly responsible for this development was the young schoolmaster, C. J. Davis who for some time had been a local preacher among the Wesleyans."
By 1866 he had sailed to Britain to study medicine with the intention to return to practise medicine in the West Indies. In London he settled in the north London suburb of Stoke Newington in the home of a Mr Holland. He became one of the House Physicians at St Bartholomew's Hospital, where in the first year of study, he gained the examiner's prize for proficiency in practical anatomy, and a junior scholarship in anatomy, physiology and chemistry.
He was an able and earnest evangelist. He preached to large congregations with much accompanying blessing in many parts of the British Isles. He records in his gospel tracts some of the locations he visited; these include Margate, Woolwich and Sheffield in England, Dunoon and Aberdeen in Scotland. At the University of Aberdeen he pursued his medical studies where he completed his degree of MD.
He cut a very striking figure and quickly drew crowds when preaching in the open air. A contemporary friend, a Harley Street doctor, A. T. Schofield, wrote that Davis was "a tall and distinguished looking man". He used to preach from a kitchen table in Union Street, Aberdeen, the result of which was the establishment of a very large assembly of Brethren in that city. In Aberdeen he delivered a series of lectures which were published the year of his death as "Aids to Believers". He also wrote a tract on "The Lord's Coming", and an evangelistic book, "The Grace of God".
His "Aids to Believers" ran through at least 18 editions in the first half of the 20th century. His final evangelistic narrative, "The Teachers Taught", is in print with a number of publishers.
Franco-Prussian War.
During the Franco-Prussian war in 1870 he volunteered his services to assist the suffering and cholera-stricken peasantry of eastern France, especially at the Battle of Sedan. He devoted himself with skill and energy to the treatment of large numbers of sick and wounded and to the establishment of soup kitchens which gave food and life to multitudes of starving people. He was an enthusiast for his soup kitchens, so that, when on one occasion, the sister of the Protestant Pastor at Sedan (Miss Goulden) told him there was not sufficient soup, he took his watch from his pocket, gained as a prize at College and sold it to pay for the immediate needs.
For this special war-work Mr and Mrs Chrimes of Moorgate, Rotherham, England, made the young physician almoner of a thousand pounds for the poor and sick among whom he laboured. Dr Davis also ran an ambulance, which was regarded as the finest one in the neighbourhood.
He also took care of several hundred wounded Bavarians in dire need of aid. It was in this final benevolent service in war-torn Europe that he assumed the honourable nickname, "The Good Black Doctor".
Death.
Two Englishwomen, Emma Maria Pearson (1828–93) and Louisa Elisabeth MacLaughlin (1836–1921), wrote this of him:
"But his enthusiasm", said "The Lancet" medical journal, "carried him beyond his strength, and, returning in an exhausted condition from a short visit to England, where he had been to seek further funds, he was attacked with small-pox, of which he died on the 27th November, at the age of twenty-eight". Davis was greatly loved for his impartial service upon his death was given a military funeral which was followed by troops of both armies, headed by the Mayor of Sedan. His tombstone records the high esteem in which he was held. He was buried in the quiet graveyard of Fond de Givonne, just outside Sedan.
In one of his tracts, the "Grace Appearing and reigning; Glory to Appear", he wrote, "go out and visit the sick – feed the hungry; and if you have the gift, explain the word of life to the poor and ignorant … All can do good in some way; with time or worldly goods … this is our time to do as receivers of grace. In health or sickness to adorn the doctrine of Christ our Saviour."
William Joseph Lowe (1839–1927), another Plymouth Brother, also visited Sedan and wrote a description of his tour.