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Hexadimethrine Bromide
Hexadimethrine bromide (commercial brand name Polybrene) is a cationic polymer with several uses. In research, it is primarily used to increase the efficiency of transduction of certain cells with viruses (such as retrovirus or lentivirus) in cell culture. Hexadimethrine bromide acts by neutralizing the charge repulsion between virions and sialic acid on the cell surface. Use of Polybrene can improve transduction efficiency 100-1000 fold although it can be toxic to some cell types. Polybrene in combination with DMSO shock is used to transfect some cell types such as NIH-3T3 and CHO. It has other uses, including a role in protein sequencing. Hexadimethrine bromide also reverses heparin anticoagulation during open-heart surgery, and it was the original reversal agents used in the 1950s and 1960s. It was replaced by protamine sulfate in 1969, after it was shown that hexadimethrine bromide could potentially cause kidney failure Kidney failure, also known as renal failure or e ...
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Cationic
An ion () is an atom or molecule with a net electrical charge. The charge of an electron is considered to be negative by convention and this charge is equal and opposite to the charge of a proton, which is considered to be positive by convention. The net charge of an ion is not zero because its total number of electrons is unequal to its total number of protons. A cation is a positively charged ion with fewer electrons than protons (e.g. K+ (potassium ion)) while an anion is a negatively charged ion with more electrons than protons (e.g. Cl− (chloride ion) and OH− (hydroxide ion)). Opposite electric charges are pulled towards one another by electrostatic force, so cations and anions attract each other and readily form ionic compounds. Ions consisting of only a single atom are termed ''monatomic ions'', ''atomic ions'' or ''simple ions'', while ions consisting of two or more atoms are termed polyatomic ions or ''molecular ions''. If only a + or − is present, it indicate ...
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Polymer
A polymer () is a chemical substance, substance or material that consists of very large molecules, or macromolecules, that are constituted by many repeat unit, repeating subunits derived from one or more species of monomers. Due to their broad spectrum of properties, both synthetic and natural polymers play essential and ubiquitous roles in everyday life. Polymers range from familiar synthetic plastics such as polystyrene to natural biopolymers such as DNA and proteins that are fundamental to biological structure and function. Polymers, both natural and synthetic, are created via polymerization of many small molecules, known as monomers. Their consequently large molecular mass, relative to small molecule compound (chemistry), compounds, produces unique physical property, physical properties including toughness, high rubber elasticity, elasticity, viscoelasticity, and a tendency to form Amorphous solid, amorphous and crystallization of polymers, semicrystalline structures rath ...
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Transduction (genetics)
Transduction is the process by which foreign DNA is introduced into a cell by a virus or viral vector. An example is the viral transfer of DNA from one bacterium to another and hence an example of horizontal gene transfer. Transduction does not require physical contact between the cell donating the DNA and the cell receiving the DNA (which occurs in conjugation), and it is DNase resistant ( transformation is susceptible to DNase). Transduction is a common tool used by molecular biologists to stably introduce a foreign gene into a host cell's genome (both bacterial and mammalian cells). Discovery (bacterial transduction) Transduction was discovered in ''Salmonella'' by Norton Zinder and Joshua Lederberg at the University of Wisconsin–Madison in 1952. In the lytic and lysogenic cycles Transduction happens through either the lytic cycle or the lysogenic cycle. When bacteriophages (viruses that infect bacteria) that are lytic infect bacterial cells, they harness the ...
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Retrovirus
A retrovirus is a type of virus that inserts a DNA copy of its RNA genome into the DNA of a host cell that it invades, thus changing the genome of that cell. After invading a host cell's cytoplasm, the virus uses its own reverse transcriptase enzyme to produce DNA from its RNA genome, the reverse of the usual pattern, thus ''retro'' (backward). The new DNA is then retroviral integration, incorporated into the host cell genome by an integrase enzyme, at which point the retroviral DNA is referred to as a provirus. The host cell then treats the viral DNA as part of its own genome, transcribing and translating the viral genes along with the cell's own genes, producing the proteins required to assemble new copies of the virus. Many retroviruses cause serious diseases in humans, other mammals, and birds. Retroviruses have many subfamilies in three basic groups. * Oncovirus, Oncoretroviruses (cancer-causing retroviruses) include human T-lymphotropic virus (HTLV) causing a type of leuk ...
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Lentivirus
''Lentivirus'' is a genus of retroviruses that cause chronic and deadly diseases characterized by long incubation periods, in humans and other mammalian species. The genus includes the human immunodeficiency virus (HIV), which causes AIDS. Lentiviruses are distributed worldwide, and are known to be hosted in apes, cows, goats, horses, cats, and sheep as well as several other mammals. Lentiviruses can integrate a significant amount of viral complementary DNA into the DNA of the host cell and can efficiently infect nondividing cells, so they are one of the most efficient methods of gene delivery. They can become endogenous, integrating their genome into the host germline genome, so that the virus is henceforth inherited by the host's descendants. Classification Five serogroups of lentiviruses are recognized, reflecting the vertebrate hosts with which they are associated (primates, sheep and goats, horses, domestic cats, and cattle). The primate lentiviruses are distinguished ...
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Cell Culture
Cell culture or tissue culture is the process by which cell (biology), cells are grown under controlled conditions, generally outside of their natural environment. After cells of interest have been Cell isolation, isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich growth medium, medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (Carbon dioxide, CO2, Oxygen, O2), and regulates the physio-chemical environment (Buffer solution, pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as a suspension culture. T ...
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Virion
A virion (plural, ''viria'' or ''virions'') is an inert virus particle capable of invading a Cell (biology), cell. Upon entering the cell, the virion disassembles and the genetic material from the virus takes control of the cell infrastructure, thus enabling the virus to Replication (virus), replicate. The genetic material (''Viral core, core'', either DNA or RNA, along with occasionally present virus core protein) inside the virion is usually enclosed in a protection shell, known as the capsid. While the terms "virus" and "virion" are occasionally confused, recently "virion" is used solely to describe the virus structure outside of cells, while the terms "virus/viral" are broader and also include biological properties such as the infectivity of a virion. Components A virion consists of one or more nucleic acid genome molecules (single-stranded or double-stranded RNA or DNA) and coatings (a capsid and possibly a viral envelope). The virion may contain other proteins (for examp ...
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Sialic Acid
Sialic acids are a class of alpha-keto acid sugars with a nine-carbon backbone. The term "sialic acid" () was first introduced by Swedish biochemist Gunnar Blix in 1952. The most common member of this group is ''N''-acetylneuraminic acid (Neu5Ac or NANA) found in animals and some prokaryotes. Sialic acids are found widely distributed in animal tissues and related forms are found to a lesser extent in other organisms like in some micro-algae, bacteria and archaea. Sialic acids are commonly part of glycoproteins, glycolipids or gangliosides, where they decorate the end of sugar chains at the surface of cells or soluble proteins. However, sialic acids have been also observed in ''Drosophila'' embryos and other insects. Generally, plants seem not to contain or display sialic acids. In humans, the brain has the highest sialic acid content, where these acids play an important role in neural transmission and ganglioside structure in synaptogenesis. More than 50 kinds of sial ...
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Cell Membrane
The cell membrane (also known as the plasma membrane or cytoplasmic membrane, and historically referred to as the plasmalemma) is a biological membrane that separates and protects the interior of a cell from the outside environment (the extracellular space). The cell membrane consists of a lipid bilayer, made up of two layers of phospholipids with cholesterols (a lipid component) interspersed between them, maintaining appropriate membrane fluidity at various temperatures. The membrane also contains membrane proteins, including integral proteins that span the membrane and serve as membrane transporters, and peripheral proteins that loosely attach to the outer (peripheral) side of the cell membrane, acting as enzymes to facilitate interaction with the cell's environment. Glycolipids embedded in the outer lipid layer serve a similar purpose. The cell membrane controls the movement of substances in and out of a cell, being selectively permeable to ions and organic mole ...
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Protein Sequencing
Protein sequencing is the practical process of determining the amino acid sequence of all or part of a protein or peptide. This may serve to identify the protein or characterize its post-translational modifications. Typically, partial sequencing of a protein provides sufficient information (one or more sequence tags) to identify it with reference to databases of protein sequences derived from the conceptual Translation (biology), translation of genes. The two major direct methods of protein sequencing are #Identification by mass spectrometry, mass spectrometry and #Edman degradation, Edman degradation using a #Protein sequenator, protein sequenator (sequencer). Mass spectrometry methods are now the most widely used for protein sequencing and identification but Edman degradation remains a valuable tool for characterizing a protein's N-terminus, ''N''-terminus. Determining amino acid composition It is often desirable to know the unordered amino acid composition of a protein prio ...
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Protamine Sulfate
Protamine sulfate is a medication that is used to reverse the effects of heparin. It is specifically used in heparin overdose, in low molecular weight heparin overdose, and to reverse the effects of heparin during delivery and heart surgery. It is given by injection into a vein. The onset of effects is typically within five minutes. Common side effects include low blood pressure, slow heart rate, allergic reactions, and vomiting. Allergic reactions may be severe and include anaphylaxis. The risk is greater in males who have had a vasectomy. While there is no evidence of harm from using during pregnancy it has not been well studied in this group. Protamine works by binding with heparin. Protamine sulfate was approved for medical use in the United States in 1969. It is on the World Health Organization's List of Essential Medicines. It was originally made from the sperm of salmon (salmine, salmon protamine). It is now mainly made using recombinant biotechnology. Medica ...
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