A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
A reverse transcriptase encoded by the POL GENE of HIV. It is a heterodimer of 66 kDa and 51 kDa subunits that are derived from a common precursor protein. The heterodimer also includes an RNAse H activity (RIBONUCLEASE H, HUMAN IMMUNODEFICIENCY VIRUS) that plays an essential role the viral replication process.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
An enzyme that synthesizes DNA on an RNA template. It is encoded by the pol gene of retroviruses and by certain retrovirus-like elements. EC
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
Inhibitors of reverse transcriptase (RNA-DIRECTED DNA POLYMERASE), an enzyme that synthesizes DNA on an RNA template.
Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)
Exfoliate neoplastic cells circulating in the blood and associated with metastasizing tumors.
RNA present in neoplastic tissue.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Methods used for detecting the amplified DNA products from the polymerase chain reaction as they accumulate instead of at the end of the reaction.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
The determination of the pattern of genes expressed at the level of GENETIC TRANSCRIPTION, under specific circumstances or in a specific cell.
Ribonucleic acid that makes up the genetic material of viruses.
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
A technique that localizes specific nucleic acid sequences within intact chromosomes, eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes.
A form of RHABDOMYOSARCOMA arising primarily in the head and neck, especially the orbit, of children below the age of 10. The cells are smaller than those of other rhabdomyosarcomas and are of two basic cell types: spindle cells and round cells. This cancer is highly sensitive to chemotherapy and has a high cure rate with multi-modality therapy. (From Holland et al., Cancer Medicine, 3d ed, p2188)
A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
A form of arboviral encephalitis (which primarily affects horses) endemic to western and central regions of NORTH AMERICA. The causative organism (ENCEPHALOMYELITIS VIRUS, WESTERN EQUINE) may be transferred to humans via the bite of mosquitoes (CULEX tarsalis and others). Clinical manifestations include headache and influenza-like symptoms followed by alterations in mentation, SEIZURES, and COMA. DEATH occurs in a minority of cases. Survivors may recover fully or be left with residual neurologic dysfunction, including PARKINSONISM, POSTENCEPHALITIC. (From Joynt, Clinical Neurology, 1996, Ch26, pp8-9)
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in neoplastic tissue.
A form of RHABDOMYOSARCOMA occurring mainly in adolescents and young adults, affecting muscles of the extremities, trunk, orbital region, etc. It is extremely malignant, metastasizing widely at an early stage. Few cures have been achieved and the prognosis is poor. "Alveolar" refers to its microscopic appearance simulating the cells of the respiratory alveolus. (Holland et al., Cancer Medicine, 3d ed, p2188)
An enzyme of the oxidoreductase class that catalyzes the reaction between L-tyrosine, L-dopa, and oxygen to yield L-dopa, dopaquinone, and water. It is a copper protein that acts also on catechols, catalyzing some of the same reactions as CATECHOL OXIDASE. EC
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
Molecular products metabolized and secreted by neoplastic tissue and characterized biochemically in cells or body fluids. They are indicators of tumor stage and grade as well as useful for monitoring responses to treatment and predicting recurrence. Many chemical groups are represented including hormones, antigens, amino and nucleic acids, enzymes, polyamines, and specific cell membrane proteins and lipids.
A positive regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
A type of chromosome aberration characterized by CHROMOSOME BREAKAGE and transfer of the broken-off portion to another location, often to a different chromosome.
Established cell cultures that have the potential to propagate indefinitely.
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
The GENETIC TRANSLATION products of the fusion between an ONCOGENE and another gene. The latter may be of viral or cellular origin.
The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.
Hybridization of a nucleic acid sample to a very large set of OLIGONUCLEOTIDE PROBES, which have been attached individually in columns and rows to a solid support, to determine a BASE SEQUENCE, or to detect variations in a gene sequence, GENE EXPRESSION, or for GENE MAPPING.
Deoxyribonucleic acid that makes up the genetic material of viruses.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Remnant of a tumor or cancer after primary, potentially curative therapy. (Dr. Daniel Masys, written communication)
Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure and transcribes DNA into RNA. It has different requirements for cations and salt than RNA polymerase I and is strongly inhibited by alpha-amanitin. EC
The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.
Elements of limited time intervals, contributing to particular results or situations.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
A prediction of the probable outcome of a disease based on a individual's condition and the usual course of the disease as seen in similar situations.
Accumulation of a drug or chemical substance in various organs (including those not relevant to its pharmacologic or therapeutic action). This distribution depends on the blood flow or perfusion rate of the organ, the ability of the drug to penetrate organ membranes, tissue specificity, protein binding. The distribution is usually expressed as tissue to plasma ratios.
Immunologic techniques based on the use of: (1) enzyme-antibody conjugates; (2) enzyme-antigen conjugates; (3) antienzyme antibody followed by its homologous enzyme; or (4) enzyme-antienzyme complexes. These are used histologically for visualizing or labeling tissue specimens.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
The relationships of groups of organisms as reflected by their genetic makeup.
A cell line derived from cultured tumor cells.
A malignant neoplasm derived from cells that are capable of forming melanin, which may occur in the skin of any part of the body, in the eye, or, rarely, in the mucous membranes of the genitalia, anus, oral cavity, or other sites. It occurs mostly in adults and may originate de novo or from a pigmented nevus or malignant lentigo. Melanomas frequently metastasize widely, and the regional lymph nodes, liver, lungs, and brain are likely to be involved. The incidence of malignant skin melanomas is rising rapidly in all parts of the world. (Stedman, 25th ed; from Rook et al., Textbook of Dermatology, 4th ed, p2445)
A malignant neoplasm arising from tenosynovial tissue of the joints and in synovial cells of tendons and bursae. The legs are the most common site, but the tumor can occur in the abdominal wall and other trunk muscles. There are two recognized types: the monophasic (characterized by sheaths of monotonous spindle cells) and the biphasic (characterized by slit-like spaces or clefts within the tumor, lined by cuboidal or tall columnar epithelial cells). These sarcomas occur most commonly in the second and fourth decades of life. (From Dorland, 27th ed; DeVita Jr et al., Cancer: Principles & Practice of Oncology, 3d ed, p1363)
The ordered rearrangement of gene regions by DNA recombination such as that which occurs normally during development.
Removal and pathologic examination of specimens in the form of small pieces of tissue from the living body.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
DNA present in neoplastic tissue.
The soft tissue filling the cavities of bones. Bone marrow exists in two types, yellow and red. Yellow marrow is found in the large cavities of large bones and consists mostly of fat cells and a few primitive blood cells. Red marrow is a hematopoietic tissue and is the site of production of erythrocytes and granular leukocytes. Bone marrow is made up of a framework of connective tissue containing branching fibers with the frame being filled with marrow cells.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
Macromolecular molds for the synthesis of complementary macromolecules, as in DNA REPLICATION; GENETIC TRANSCRIPTION of DNA to RNA, and GENETIC TRANSLATION of RNA into POLYPEPTIDES.
A malignant tumor of the bone which always arises in the medullary tissue, occurring more often in cylindrical bones. The tumor occurs usually before the age of 20, about twice as frequently in males as in females.
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
A class of fibrous proteins or scleroproteins that represents the principal constituent of EPIDERMIS; HAIR; NAILS; horny tissues, and the organic matrix of tooth ENAMEL. Two major conformational groups have been characterized, alpha-keratin, whose peptide backbone forms a coiled-coil alpha helical structure consisting of TYPE I KERATIN and a TYPE II KERATIN, and beta-keratin, whose backbone forms a zigzag or pleated sheet structure. alpha-Keratins have been classified into at least 20 subtypes. In addition multiple isoforms of subtypes have been found which may be due to GENE DUPLICATION.
Proteins prepared by recombinant DNA technology.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.
The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
A negative regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
A type of IN SITU HYBRIDIZATION in which target sequences are stained with fluorescent dye so their location and size can be determined using fluorescence microscopy. This staining is sufficiently distinct that the hybridization signal can be seen both in metaphase spreads and in interphase nuclei.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.
Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).
Non-antibody proteins secreted by inflammatory leukocytes and some non-leukocytic cells, that act as intercellular mediators. They differ from classical hormones in that they are produced by a number of tissue or cell types rather than by specialized glands. They generally act locally in a paracrine or autocrine rather than endocrine manner.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
An essential ribonucleoprotein reverse transcriptase that adds telomeric DNA to the ends of eukaryotic CHROMOSOMES.
All of the processes involved in increasing CELL NUMBER including CELL DIVISION.
The type species of LENTIVIRUS and the etiologic agent of AIDS. It is characterized by its cytopathic effect and affinity for the T4-lymphocyte.
A process whereby multiple RNA transcripts are generated from a single gene. Alternative splicing involves the splicing together of other possible sets of EXONS during the processing of some, but not all, transcripts of the gene. Thus a particular exon may be connected to any one of several alternative exons to form a mature RNA. The alternative forms of mature MESSENGER RNA produce PROTEIN ISOFORMS in which one part of the isoforms is common while the other parts are different.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
Short fragments of DNA or RNA that are used to alter the function of target RNAs or DNAs to which they hybridize.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
A DNA-dependent DNA polymerase characterized in prokaryotes and may be present in higher organisms. It has both 3'-5' and 5'-3' exonuclease activity, but cannot use native double-stranded DNA as template-primer. It is not inhibited by sulfhydryl reagents and is active in both DNA synthesis and repair. EC
Tumors or cancer of the SKIN.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action during the developmental stages of an organism.
Proteins, glycoprotein, or lipoprotein moieties on surfaces of tumor cells that are usually identified by monoclonal antibodies. Many of these are of either embryonic or viral origin.
A factor synthesized in a wide variety of tissues. It acts synergistically with TGF-alpha in inducing phenotypic transformation and can also act as a negative autocrine growth factor. TGF-beta has a potential role in embryonal development, cellular differentiation, hormone secretion, and immune function. TGF-beta is found mostly as homodimer forms of separate gene products TGF-beta1, TGF-beta2 or TGF-beta3. Heterodimers composed of TGF-beta1 and 2 (TGF-beta1.2) or of TGF-beta2 and 3 (TGF-beta2.3) have been isolated. The TGF-beta proteins are synthesized as precursor proteins.
Studies which start with the identification of persons with a disease of interest and a control (comparison, referent) group without the disease. The relationship of an attribute to the disease is examined by comparing diseased and non-diseased persons with regard to the frequency or levels of the attribute in each group.
Sudden increase in the incidence of a disease. The concept includes EPIDEMICS and PANDEMICS.
Methods which attempt to express in replicable terms the extent of the neoplasm in the patient.
One of the mechanisms by which CELL DEATH occurs (compare with NECROSIS and AUTOPHAGOCYTOSIS). Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. It is characterized by distinctive morphologic changes in the nucleus and cytoplasm, chromatin cleavage at regularly spaced sites, and the endonucleolytic cleavage of genomic DNA; (DNA FRAGMENTATION); at internucleosomal sites. This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.
Simultaneous resistance to several structurally and functionally distinct drugs.
The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.
A soluble factor produced by MONOCYTES; MACROPHAGES, and other cells which activates T-lymphocytes and potentiates their response to mitogens or antigens. Interleukin-1 is a general term refers to either of the two distinct proteins, INTERLEUKIN-1ALPHA and INTERLEUKIN-1BETA. The biological effects of IL-1 include the ability to replace macrophage requirements for T-cell activation.
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
Serum glycoprotein produced by activated MACROPHAGES and other mammalian MONONUCLEAR LEUKOCYTES. It has necrotizing activity against tumor cell lines and increases ability to reject tumor transplants. Also known as TNF-alpha, it is only 30% homologous to TNF-beta (LYMPHOTOXIN), but they share TNF RECEPTORS.
The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.
Mature LYMPHOCYTES and MONOCYTES transported by the blood to the body's extravascular space. They are morphologically distinguishable from mature granulocytic leukocytes by their large, non-lobed nuclei and lack of coarse, heavily stained cytoplasmic granules.
The outer covering of the body that protects it from the environment. It is composed of the DERMIS and the EPIDERMIS.
Naturally occurring or experimentally induced animal diseases with pathological processes sufficiently similar to those of human diseases. They are used as study models for human diseases.
Observation of a population for a sufficient number of persons over a sufficient number of years to generate incidence or mortality rates subsequent to the selection of the study group.
Tumors or cancer of the human BREAST.
Epidermal cells which synthesize keratin and undergo characteristic changes as they move upward from the basal layers of the epidermis to the cornified (horny) layer of the skin. Successive stages of differentiation of the keratinocytes forming the epidermal layers are basal cell, spinous or prickle cell, and the granular cell.
An acute viral infection in humans involving the respiratory tract. It is marked by inflammation of the NASAL MUCOSA; the PHARYNX; and conjunctiva, and by headache and severe, often generalized, myalgia.
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
The relationship between the dose of an administered drug and the response of the organism to the drug.
A strain of albino rat developed at the Wistar Institute that has spread widely at other institutions. This has markedly diluted the original strain.
A 170-kDa transmembrane glycoprotein from the superfamily of ATP-BINDING CASSETTE TRANSPORTERS. It serves as an ATP-dependent efflux pump for a variety of chemicals, including many ANTINEOPLASTIC AGENTS. Overexpression of this glycoprotein is associated with multidrug resistance (see DRUG RESISTANCE, MULTIPLE).
Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs.
A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms. It may be present in higher organisms and has an intrinsic molecular activity only 5% of that of DNA Polymerase I. This polymerase has 3'-5' exonuclease activity, is effective only on duplex DNA with gaps or single-strand ends of less than 100 nucleotides as template, and is inhibited by sulfhydryl reagents. EC
A class of statistical procedures for estimating the survival function (function of time, starting with a population 100% well at a given time and providing the percentage of the population still well at later times). The survival analysis is then used for making inferences about the effects of treatments, prognostic factors, exposures, and other covariates on the function.
The male gonad containing two functional parts: the SEMINIFEROUS TUBULES for the production and transport of male germ cells (SPERMATOGENESIS) and the interstitial compartment containing LEYDIG CELLS that produce ANDROGENS.
A subtype of INFLUENZA A VIRUS with the surface proteins hemagglutinin 1 and neuraminidase 1. The H1N1 subtype was responsible for the Spanish flu pandemic of 1918.
A neoplasm characterized by abnormalities of the lymphoid cell precursors leading to excessive lymphoblasts in the marrow and other organs. It is the most common cancer in children and accounts for the vast majority of all childhood leukemias.
A cytokine that stimulates the growth and differentiation of B-LYMPHOCYTES and is also a growth factor for HYBRIDOMAS and plasmacytomas. It is produced by many different cells including T-LYMPHOCYTES; MONOCYTES; and FIBROBLASTS.
In screening and diagnostic tests, the probability that a person with a positive test is a true positive (i.e., has the disease), is referred to as the predictive value of a positive test; whereas, the predictive value of a negative test is the probability that the person with a negative test does not have the disease. Predictive value is related to the sensitivity and specificity of the test.
Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.
Small double-stranded, non-protein coding RNAs, 21-25 nucleotides in length generated from single-stranded microRNA gene transcripts by the same RIBONUCLEASE III, Dicer, that produces small interfering RNAs (RNA, SMALL INTERFERING). They become part of the RNA-INDUCED SILENCING COMPLEX and repress the translation (TRANSLATION, GENETIC) of target RNA by binding to homologous 3'UTR region as an imperfect match. The small temporal RNAs (stRNAs), let-7 and lin-4, from C. elegans, are the first 2 miRNAs discovered, and are from a class of miRNAs involved in developmental timing.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
A ribonuclease that specifically cleaves the RNA moiety of RNA:DNA hybrids. It has been isolated from a wide variety of prokaryotic and eukaryotic organisms as well as RETROVIRUSES.
Different forms of a protein that may be produced from different GENES, or from the same gene by ALTERNATIVE SPLICING.
Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.
They are oval or bean shaped bodies (1 - 30 mm in diameter) located along the lymphatic system.
Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen.
Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules.
Cells that line the inner and outer surfaces of the body by forming cellular layers (EPITHELIUM) or masses. Epithelial cells lining the SKIN; the MOUTH; the NOSE; and the ANAL CANAL derive from ectoderm; those lining the RESPIRATORY SYSTEM and the DIGESTIVE SYSTEM derive from endoderm; others (CARDIOVASCULAR SYSTEM and LYMPHATIC SYSTEM) derive from mesoderm. Epithelial cells can be classified mainly by cell shape and function into squamous, glandular and transitional epithelial cells.
Methods for maintaining or growing CELLS in vitro.
Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.
Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction.
A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms but may be present in higher organisms. Use also for a more complex form of DNA polymerase III designated as DNA polymerase III* or pol III* which is 15 times more active biologically than DNA polymerase I in the synthesis of DNA. This polymerase has both 3'-5' and 5'-3' exonuclease activities, is inhibited by sulfhydryl reagents, and has the same template-primer dependence as pol II. EC
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
The fission of a CELL. It includes CYTOKINESIS, when the CYTOPLASM of a cell is divided, and CELL NUCLEUS DIVISION.
Tumors or cancer of the LIVER.
Polymorphic cells that form cartilage.
A selective increase in the number of copies of a gene coding for a specific protein without a proportional increase in other genes. It occurs naturally via the excision of a copy of the repeating sequence from the chromosome and its extrachromosomal replication in a plasmid, or via the production of an RNA transcript of the entire repeating sequence of ribosomal RNA followed by the reverse transcription of the molecule to produce an additional copy of the original DNA sequence. Laboratory techniques have been introduced for inducing disproportional replication by unequal crossing over, uptake of DNA from lysed cells, or generation of extrachromosomal sequences from rolling circle replication.
Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
Small double-stranded, non-protein coding RNAs (21-31 nucleotides) involved in GENE SILENCING functions, especially RNA INTERFERENCE (RNAi). Endogenously, siRNAs are generated from dsRNAs (RNA, DOUBLE-STRANDED) by the same ribonuclease, Dicer, that generates miRNAs (MICRORNAS). The perfect match of the siRNAs' antisense strand to their target RNAs mediates RNAi by siRNA-guided RNA cleavage. siRNAs fall into different classes including trans-acting siRNA (tasiRNA), repeat-associated RNA (rasiRNA), small-scan RNA (scnRNA), and Piwi protein-interacting RNA (piRNA) and have different specific gene silencing functions.
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
A set of statistical methods used to group variables or observations into strongly inter-related subgroups. In epidemiology, it may be used to analyze a closely grouped series of events or cases of disease or other health-related phenomenon with well-defined distribution patterns in relation to time or place or both.
Deoxyribonucleic acid that makes up the genetic material of protozoa.
Electrophoresis in which agar or agarose gel is used as the diffusion medium.
Glycoproteins found on the membrane or surface of cells.
Measurable and quantifiable biological parameters (e.g., specific enzyme concentration, specific hormone concentration, specific gene phenotype distribution in a population, presence of biological substances) which serve as indices for health- and physiology-related assessments, such as disease risk, psychiatric disorders, environmental exposure and its effects, disease diagnosis, metabolic processes, substance abuse, pregnancy, cell line development, epidemiologic studies, etc.
Substances that inhibit or prevent the proliferation of NEOPLASMS.
A class of statistical methods applicable to a large set of probability distributions used to test for correlation, location, independence, etc. In most nonparametric statistical tests, the original scores or observations are replaced by another variable containing less information. An important class of nonparametric tests employs the ordinal properties of the data. Another class of tests uses information about whether an observation is above or below some fixed value such as the median, and a third class is based on the frequency of the occurrence of runs in the data. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed, p1284; Corsini, Concise Encyclopedia of Psychology, 1987, p764-5)
Agents used to treat AIDS and/or stop the spread of the HIV infection. These do not include drugs used to treat symptoms or opportunistic infections associated with AIDS.
A member of the CXC chemokine family that plays a role in the regulation of the acute inflammatory response. It is secreted by variety of cell types and induces CHEMOTAXIS of NEUTROPHILS and other inflammatory cells.
A malignant epithelial tumor with a glandular organization.
The regular and simultaneous occurrence in a single interbreeding population of two or more discontinuous genotypes. The concept includes differences in genotypes ranging in size from a single nucleotide site (POLYMORPHISM, SINGLE NUCLEOTIDE) to large nucleotide sequences visible at a chromosomal level.
Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.
A cytokine produced by a variety of cell types, including T-LYMPHOCYTES; MONOCYTES; DENDRITIC CELLS; and EPITHELIAL CELLS that exerts a variety of effects on immunoregulation and INFLAMMATION. Interleukin-10 combines with itself to form a homodimeric molecule that is the biologically active form of the protein.
The worsening of a disease over time. This concept is most often used for chronic and incurable diseases where the stage of the disease is an important determinant of therapy and prognosis.
An infant during the first month after birth.
Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.
The major interferon produced by mitogenically or antigenically stimulated LYMPHOCYTES. It is structurally different from TYPE I INTERFERON and its major activity is immunoregulation. It has been implicated in the expression of CLASS II HISTOCOMPATIBILITY ANTIGENS in cells that do not normally produce them, leading to AUTOIMMUNE DISEASES.
A dideoxynucleoside compound in which the 3'-hydroxy group on the sugar moiety has been replaced by an azido group. This modification prevents the formation of phosphodiester linkages which are needed for the completion of nucleic acid chains. The compound is a potent inhibitor of HIV replication, acting as a chain-terminator of viral DNA during reverse transcription. It improves immunologic function, partially reverses the HIV-induced neurological dysfunction, and improves certain other clinical abnormalities associated with AIDS. Its principal toxic effect is dose-dependent suppression of bone marrow, resulting in anemia and leukopenia.
Tumors or cancer of the COLON or the RECTUM or both. Risk factors for colorectal cancer include chronic ULCERATIVE COLITIS; FAMILIAL POLYPOSIS COLI; exposure to ASBESTOS; and irradiation of the CERVIX UTERI.
Differentiation antigens residing on mammalian leukocytes. CD stands for cluster of differentiation, which refers to groups of monoclonal antibodies that show similar reactivity with certain subpopulations of antigens of a particular lineage or differentiation stage. The subpopulations of antigens are also known by the same CD designation.
The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM.
Transfer of a neoplasm from its primary site to lymph nodes or to distant parts of the body by way of the lymphatic system.
Products of proto-oncogenes. Normally they do not have oncogenic or transforming properties, but are involved in the regulation or differentiation of cell growth. They often have protein kinase activity.
The transfer of a neoplasm from one organ or part of the body to another remote from the primary site.
The ability of viruses to resist or to become tolerant to chemotherapeutic agents or antiviral agents. This resistance is acquired through gene mutation.
The original member of the family of endothelial cell growth factors referred to as VASCULAR ENDOTHELIAL GROWTH FACTORS. Vascular endothelial growth factor-A was originally isolated from tumor cells and referred to as "tumor angiogenesis factor" and "vascular permeability factor". Although expressed at high levels in certain tumor-derived cells it is produced by a wide variety of cell types. In addition to stimulating vascular growth and vascular permeability it may play a role in stimulating VASODILATION via NITRIC OXIDE-dependent pathways. Alternative splicing of the mRNA for vascular endothelial growth factor A results in several isoforms of the protein being produced.
The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
Evaluation undertaken to assess the results or consequences of management and procedures used in combating disease in order to determine the efficacy, effectiveness, safety, and practicability of these interventions in individual cases or series.
The rate dynamics in chemical or physical systems.
The phosphate esters of DIDEOXYNUCLEOSIDES.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. The enzyme functions in the nucleolar structure and transcribes DNA into RNA. It has different requirements for cations and salts than RNA polymerase II and III and is not inhibited by alpha-amanitin. EC
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure where it transcribes DNA into RNA. It has specific requirements for cations and salt and has shown an intermediate sensitivity to alpha-amanitin in comparison to RNA polymerase I and II. EC
A species of ALPHARETROVIRUS causing anemia in fowl.
Tumors or cancer of the LUNG.
Proteins encoded by homeobox genes (GENES, HOMEOBOX) that exhibit structural similarity to certain prokaryotic and eukaryotic DNA-binding proteins. Homeodomain proteins are involved in the control of gene expression during morphogenesis and development (GENE EXPRESSION REGULATION, DEVELOPMENTAL).
Tumors or cancer of the PROSTATE.
Lymphocytes responsible for cell-mediated immunity. Two types have been identified - cytotoxic (T-LYMPHOCYTES, CYTOTOXIC) and helper T-lymphocytes (T-LYMPHOCYTES, HELPER-INDUCER). They are formed when lymphocytes circulate through the THYMUS GLAND and differentiate to thymocytes. When exposed to an antigen, they divide rapidly and produce large numbers of new T cells sensitized to that antigen.
The largest of polypeptide chains comprising immunoglobulins. They contain 450 to 600 amino acid residues per chain, and have molecular weights of 51-72 kDa.
Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)
A DNA repair enzyme that catalyzes DNA synthesis during base excision DNA repair. EC
The local recurrence of a neoplasm following treatment. It arises from microscopic cells of the original neoplasm that have escaped therapeutic intervention and later become clinically visible at the original site.
Either of the pair of organs occupying the cavity of the thorax that effect the aeration of the blood.
The movement of cells from one location to another. Distinguish from CYTOKINESIS which is the process of dividing the CYTOPLASM of a cell.
Studies used to test etiologic hypotheses in which inferences about an exposure to putative causal factors are derived from data relating to characteristics of persons under study or to events or experiences in their past. The essential feature is that some of the persons under study have the disease or outcome of interest and their characteristics are compared with those of unaffected persons.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
Biochemical identification of mutational changes in a nucleotide sequence.
Phosphate esters of THYMIDINE in N-glycosidic linkage with ribose or deoxyribose, as occurs in nucleic acids. (From Dorland, 28th ed, p1154)
The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle.
Lipid-containing polysaccharides which are endotoxins and important group-specific antigens. They are often derived from the cell wall of gram-negative bacteria and induce immunoglobulin secretion. The lipopolysaccharide molecule consists of three parts: LIPID A, core polysaccharide, and O-specific chains (O ANTIGENS). When derived from Escherichia coli, lipopolysaccharides serve as polyclonal B-cell mitogens commonly used in laboratory immunology. (From Dorland, 28th ed)
A mutation caused by the substitution of one nucleotide for another. This results in the DNA molecule having a change in a single base pair.

Alternative sulfonylurea receptor expression defines metabolic sensitivity of K-ATP channels in dopaminergic midbrain neurons. (1/55562)

ATP-sensitive potassium (K-ATP) channels couple the metabolic state to cellular excitability in various tissues. Several isoforms of the K-ATP channel subunits, the sulfonylurea receptor (SUR) and inwardly rectifying K channel (Kir6.X), have been cloned, but the molecular composition and functional diversity of native neuronal K-ATP channels remain unresolved. We combined functional analysis of K-ATP channels with expression profiling of K-ATP subunits at the level of single substantia nigra (SN) neurons in mouse brain slices using an RT-multiplex PCR protocol. In contrast to GABAergic neurons, single dopaminergic SN neurons displayed alternative co-expression of either SUR1, SUR2B or both SUR isoforms with Kir6.2. Dopaminergic SN neurons expressed alternative K-ATP channel species distinguished by significant differences in sulfonylurea affinity and metabolic sensitivity. In single dopaminergic SN neurons, co-expression of SUR1 + Kir6.2, but not of SUR2B + Kir6.2, correlated with functional K-ATP channels highly sensitive to metabolic inhibition. In contrast to wild-type, surviving dopaminergic SN neurons of homozygous weaver mouse exclusively expressed SUR1 + Kir6.2 during the active period of dopaminergic neurodegeneration. Therefore, alternative expression of K-ATP channel subunits defines the differential response to metabolic stress and constitutes a novel candidate mechanism for the differential vulnerability of dopaminergic neurons in response to respiratory chain dysfunction in Parkinson's disease.  (+info)

Anopheles gambiae Ag-STAT, a new insect member of the STAT family, is activated in response to bacterial infection. (2/55562)

A new insect member of the STAT family of transcription factors (Ag-STAT) has been cloned from the human malaria vector Anopheles gambiae. The domain involved in DNA interaction and the SH2 domain are well conserved. Ag-STAT is most similar to Drosophila D-STAT and to vertebrate STATs 5 and 6, constituting a proposed ancient class A of the STAT family. The mRNA is expressed at all developmental stages, and the protein is present in hemocytes, pericardial cells, midgut, skeletal muscle and fat body cells. There is no evidence of transcriptional activation following bacterial challenge. However, bacterial challenge results in nuclear translocation of Ag-STAT protein in fat body cells and induction of DNA-binding activity that recognizes a STAT target site. In vitro treatment with pervanadate (vanadate and H2O2) translocates Ag-STAT to the nucleus in midgut epithelial cells. This is the first evidence of direct participation of the STAT pathway in immune responses in insects.  (+info)

Expression of novel alternatively spliced isoforms of the oct-1 transcription factor. (3/55562)

Analysis of the alternatively spliced isoforms of the human and mouse oct-1 genes, combined with their exon-intron structure, show a high level of evolutionary conservation between these two species. The differential expression of several oct-1 isoforms was examined by reverse transcription-polymerase chain reaction performed on the 3' region of the murine oct-1 cDNA. Variations in the relative levels and patterns of expression of the isoforms were found among different tissues. Three novel isoforms originating from the 3'-distal region of oct-1, were isolated and sequenced: Two were derived from testis, and one from myeloma cells. Splicing out of different exons as revealed in the structure of these isoforms results in reading frameshifts that presumably lead to the expression of shortened Oct-1 proteins, with distinct C-terminal tails. Altogether, six out of the eight known murine oct-1 isoforms may have distinct C-termini, implying that these multiple tails have different functional roles in cellular differentiation and physiology.  (+info)

Chemokine mRNA expression in gastric mucosa is associated with Helicobacter pylori cagA positivity and severity of gastritis. (4/55562)

AIM: To investigate the association between the quantity of gastric chemokine mRNA expression, severity of gastritis, and cagA positivity in Helicobacter pylori associated gastritis. METHODS: In 83 dyspeptic patients, antral and corpus biopsies were taken for semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and histological grading of gastritis. Gastritis was evaluated by visual analogue scales. Quantities of chemokine (IL-8, GRO alpha, ENA-78, RANTES, MCP-1) RT-PCR products were compared with G3PDH products. Each sample was also evaluated for the presence of cagA and ureA mRNA by RT-PCR. RESULTS: mRNA expression of all five chemokines was significantly greater in H pylori positive than in H pylori negative mucosa. In H pylori positive patients, in the antrum C-X-C chemokine mRNA expression was significantly greater in cagA positive patients than in cagA negative patients, but there were no significant differences in C-C chemokine mRNA expression. In H pylori positive patients, chemokine mRNA expression in the corpus was less than in the antrum. In contrast to the antrum, only GRO alpha mRNA expression was significantly greater in cagA positive infection. Polymorphonuclear cell infiltration was correlated with C-X-C chemokine mRNA expression. Significant correlations were also found between bacterial density and C-X-C chemokine mRNA expression. CONCLUSIONS: In H pylori infection, C-X-C chemokines may play a primary role in active gastritis. Infection with cagA positive H pylori induces greater gastric chemokine mRNA expression in the antral mucosa, which may be relevant to the increased mucosal damage associated with cagA positive H pylori infection.  (+info)

The role of alternative splicing of the adhesion molecule, CD44, in lymphoid malignancy. (5/55562)

AIM: To investigate the expression of CD44 isoforms containing variant exon 6 (v6) in a well characterised cohort of patients with non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukaemia (CLL), and to correlate this with phenotype and disease course. METHODS: Cryostat sections of OCT embedded diagnostic nodal material from NHL patients and cryopreserved mononuclear preparations from CLL patients were used as sources of RNA. After reverse transcription, PCR was carried out with amplimers positioned at either side of the variant exon insertion site to amplify all possible CD44 isoforms. Those isoforms containing v6 were identified after Southern blotting and hybridisation with a radiolabelled oligonucleotide. RESULTS: Of 32 NHL samples analysed, 16 did not express CD44 isoforms containing v6, six expressed an isoform containing exon v6 alone, and 10 expressed v6 long isoforms which contained exon v6 in addition to other variant exons. These data did not correlate with lymphoma classification, disease staging, or the presence or absence of extranodal disease. However, those patients expressing v6 long CD44 isoforms had a worse overall survival than those that did not. The plateau of the survival curves was 50% compared with 82%. No v6 long isoforms were detected in the 21 CLL samples investigated. CONCLUSIONS: The expression of v6 long CD44 isoforms is associated with aggressive disease in NHL, independent of grade, stage, or presence of extranodal disease.  (+info)

Transcriptional regulation and induction of apoptosis: implications for the use of monomeric p53 variants in gene therapy. (6/55562)

The p53 tumour suppressor protein is a transcriptional activator, which can induce cell cycle arrest and apoptosis. p53 Gene mutations occur in more than 50% of all human tumours. Reintroduction of wild-type p53 but also of oligomerisation-independent p53 variants into tumour cells by gene transfer methods has been considered. We have investigated the biological properties of two carboxy-terminal deletion mutants of p53, p53 delta 300 (comprising amino acids 1-300) and p53 delta 326 (amino acids 1-326), to evaluate their potential deployment in gene therapy. Transactivation was measured in transiently transfected HeLa and SKBR3 cells. Both monomeric variants showed reduced activities compared with wild-type p53. Individual promoters were differently affected. In contrast to wild-type p53, monomeric variants were not able to induce apoptosis. We also provided wild-type p53 and p53 delta 326 with tetracycline-regulated promoters and stably introduced these constructs into Saos2 and SKBR3 cells. Upon induction, wild-type p53 expressing cells, but not p53 delta 326 expressing cells underwent apoptosis. Consistently, only wild-type p53 expressing cells accumulated p21/waf1/cip1 mRNA and protein and showed increased bax, Gadd45 and mdm2 mRNA. Neither wild-type p53 nor p53 delta 326 repressed the transcription of the IGF-1R gene in these cell lines. We conclude that the transactivation potential of monomeric, carboxy-terminally truncated p53 is not sufficient to cause induction of the endogenous target genes which trigger apoptosis.  (+info)

Astrocyte-specific expression of tyrosine hydroxylase after intracerebral gene transfer induces behavioral recovery in experimental parkinsonism. (7/55562)

Parkinson's disease is a neurodegenerative disorder characterized by the depletion of dopamine in the caudate putamen. Dopamine replacement with levodopa, a precursor of the neurotransmitter, is presently the most common treatment for this disease. However, in an effort to obtain better therapeutic results, tissue or cells that synthesize catecholamines have been grafted into experimental animals and human patients. In this paper, we present a novel technique to express tyrosine hydroxylase (TH) in the host's own astrocytes. This procedure uses a transgene in which the expression of a TH cDNA is under the control of a glial fibrillary acidic protein (GFAP) promoter, which confers astrocyte-specific expression and also increases its activity in response to brain injury. The method was tested in a rat model of Parkinson's disease produced by lesioning the striatum with 6-hydroxydopamine. Following microinjection of the transgene into the denervated striatum as a DNA-liposome complex, expression of the transgene was detected by RT-PCR and TH protein was observed specifically in astrocytes by using double-labeling immunofluorescence for GFAP and TH coupled with laser confocal microscopy. Efficacy was demonstrated by significant behavioral recovery, as assessed by a decrease in the pharmacologically induced turning behavior generated by the unilateral denervation of the rat striatum. These results suggest this is a valuable technique to express molecules of therapeutic interest in the brain.  (+info)

Increased expression of fibroblast growth factor 8 in human breast cancer. (8/55562)

Fibroblast growth factor 8 (FGF8) is an important developmental protein which is oncogenic and able to cooperate with wnt-1 to produce mouse mammary carcinoma. The level of expression of FGF8 mRNA was measured in 68 breast cancers and 24 non-malignant breast tissues. Elevated levels of FGF8 mRNA were found in malignant compared to non-malignant breast tissues with significantly more malignant tissues expressing FGF8 (P=0.019) at significantly higher levels (P=0.031). In situ hybridization of breast cancer tissues and analysis of purified populations of normal epithelial cells and breast cancer cell lines showed that malignant epithelial cells expressed FGF8 mRNA at high levels compared to non-malignant epithelial and myoepithelial cells and fibroblasts. Although two of the receptors which FGF8 binds to (FGFR2-IIIc, FGFR3-IIIc) are not expressed in breast cancer cells, an autocrine activation loop is possible since expression of fibroblast growth factor receptor (FGFR) 4 and FGFR1 are retained in malignant epithelial cells. This is the first member of the FGF family to have increased expression in breast cancer and a potential autocrine role in its progression.  (+info)

Fingerprint Dive into the research topics of Feline coronavirus quantitative reverse-transcriptase polymerase chain reaction on effusion samples in cats with and without feline infectious peritonitis. Together they form a unique fingerprint. ...
Computed tomography (CT) of the chest is commonly used in the clinical management and assessment of complications of pneumonia caused by the novel coronavirus-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-, as well as to exclude alternative diagnoses(1). Although chest CT plays an important role, it should not be utilized as a screening tool and it can not be used by itself to confirm or exclude the diagnosis of this entity known as COVID-19(2,3), first described in the city of Wuhan, in the province of Hubei, in China, and declared as a pandemic by the World Health Organization in March 2020(4).. The diagnosis of COVID-19 is confirmed by real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and partial or complete sequencing of the viral genome from nasopharyngeal aspirates, nasal and oral swabs, sputum or tracheal or bronchial lavage(5). The vast majority of patients with SARS-CoV-2 is asymptomatic, but the spectrum of clinical presentation is broad including ...
A quantitative real time reverse-transcription polymerase chain reaction (qRT-PCR) assay with specific primers recommended by the World Health Organization (WHO) has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin (HA) genes of the pandemic H1N1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold
TY - JOUR. T1 - Quantitative RT-PCR assay for HPV infection in cultured cells. AU - Culp, Timothy D.. AU - Christensen, Neil D.. PY - 2003/8/1. Y1 - 2003/8/1. N2 - Early events in the life cycle of the human papillomaviruses (HPV) have been difficult to investigate due to both the scarcity of authentic HPV virions and limitations in assays to detect and quantify nonpermissive infections in monolayer cell culture. We have developed a quantitative reverse transcription-PCR (QRT-PCR) assay for the E1 ^ E4 transcript of HPV-11. This assay is both sensitive, and capable of differentiating between infections caused by a wide range of virus input. The QRT-PCR assay measured accurately the relative amount of viral transcripts present in samples during validation experiments using RNAs from three cell lines. Infections in all three cell lines, using titrations of HPV-11 virions ranging from 20 to 600 particles per cell, produced linear expression profiles suggesting that these multiplicities of infection ...
TY - JOUR. T1 - Real-time quantitative reverse transcriptase polymerase chain reaction.. AU - Fan, Hongxin. AU - Robetorye, Ryan S.. PY - 2010. Y1 - 2010. N2 - The real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) has become the method of choice for the quantification of specific mRNAs. This method is fast, extremely sensitive, and accurate, requires only very small amounts of input RNA, and is relatively simple to perform. These characteristics have made it the method of choice for minimal residual disease monitoring such as in chronic myelogenous leukemia (CML). CML comprises approximately 20% of all leukemias and is characterized by a balanced (9;22) chromosomal translocation that results in the formation of a chimeric gene comprised of the BCR (breakpoint cluster region) gene and the ABL oncogene (BCR-ABL fusion gene). The chimeric gene encodes a fusion protein with constitutively increased tyrosine kinase activity, resulting in growth factor-independent ...
PubMedID: 23777485 | Quantitative expression analysis and prognostic significance of the BCL2-associated X gene in nasopharyngeal carcinoma: a retrospective cohort study. | BMC cancer | 1/1/2013
In final result, this analysis implies a surrounding function for coinfecting infections in clients introducing with SARI and highlights the crucial role of viral coinfection. Persisted use of the rRT-PCR multiplex assay in combination with the SARI surveillance program will boost our capability to find blood circulation of respiratory trojans in clients hospitalized for SARI and aid explain the factor of these respiratory system viruses among individuals with SARI in Southwest Photography equipment. Although there happen to be increasing problems about the prospective for A(H1N1)pdm09 to reassort with pre-existing real human influenza infections and give go up to a very transmissible or pathogenic pathogen [22], no blended infections have been noticed with either subtypes in individuals with SARI in this analysis.. A lymphocytic meningitis had been current in 6 of 21 HIV -seropositive people but in zero of the HIV -seronegative patients. Perivascular infiltrates consisting of lymphocytes and ...
The dysregulated circular RNAs (circRNAs) are relevant to the development of osteoarthritis (OA). The circRNA serpin family E member 2 (circSERPINE2) is dysregulated in OA, while the role and mechanism of circSERPINE2 in OA are largely unknown. The aim of our research is to explore how and whether circSERPINE2 regulates interleukin-1β (IL-1β)-caused chondrocyte damage in OA. In the present study, the chondrocytes (CHON-001 cells) were exposed to IL-1β to mimic the injury in OA. CircSERPINE2, microRNA-495 (miR-495) and transforming growth factor-β receptor 2 (TGFBR2) abundances were detected via quantitative reverse-transcription polymerase chain reaction (qRT-PCR) or Western blot. Cell apoptosis was assessed via viability, apoptotic rate and caspase-3 activity. Extracellular matrix was investigated by levels of Sry-type high-mobility-group box 9 (SOX9), collagen type II α 1 (COL2A1) and Aggrecan using Western blot. The interaction among circSERPINE2, miR-495 and TGFBR2 was assessed via ...
ONC201/TIC10 is a first-in-class small molecule inducer of TRAIL that causes early activation of the integrated stress response. Its promising safety profile and broad-spectrum efficacy in vitro have been confirmed in Phase I/II trials in several advanced malignancies. Binding and reporter assays have shown that ONC201 is a selective antagonist of the dopamine D2-like receptors, specifically, DRD2 and DRD3. We hypothesized that ONC201s interaction with DRD2 plays a role in ONC201s anticancer effects. Using cBioportal and quantitative reverse-transcription polymerase chain reaction analyses, we confirmed that DRD2 is expressed in different cancer cell types in a cell type-specific manner ...
Rani S, ODriscoll L., Reverse-transcriptase polymerase chain reaction to detect extracellular mRNAs., Methods Mol Biol., 784, 2011, 15 - 25 ...
The Deoxy+ OneStep RT-PCR Kit is a ready-to-use master mix which eliminates the need for optimization of reaction and cycling conditions for one-step RT-PCR. The reaction can be prepared by simply adding template RNA and primers to the master mix. The use of Yeasterns Hotstart DNA polymerase and Deoxy+ HiSpec RT enables reliable real-time RT-PCR quantification on any real-time PCR machines. Since it is a one-tube reaction, the procedure makes high-throughput analysis possible.. After reverse transcription, reactions are heated to 95°C for 10 minutes to inactivate the reverse transcriptase and simultaneously activate HotStart Taq DNA polymerase. This hot start to the PCR eliminates any nonspecific amplification products such as primer-dimers and reduces background smear, ensuring highly sensitive and reproducible RT-PCR.. ...
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How can one calculate the efficiency of isolated total RNA from different tissues treated at the same time, when it comes to doing quantitative real-time RT-PCR analysis afterwards? If one compare the real-time expression of a certain gene in different tissues, using housekeeping genes and an external standard reference gene.. How can one be sure that the possible difference in the expression pattern is not due to difference in the isolation efficiency?. ...
subject: discussion on Quantitative Competitive RT-PCR dear netters, I am busy writing a discussion for a report I hope graduate on. So at the moment I am looking for some additional comments and insights on Quantitative Competitive RT-PCR that will help me getting a more thorough discussion. Here are some of the questions I am taking in account: What are the strong and weak points in the QC RT-PCR system (internal control: insertion of approx. 4bp; 5 fluorescent labeled primer; genescan analysis/ABI 370)? What is the best way to validate the method you are trying to set up. And are the results obtained with this method reliable/reproducible enough to get accepted by reviewers. Is there some strong literature discussing the system including the disadvantages of it. Thanks in advance, David E. Sluijs Pathology, Academic Hospital Utrecht. please mail to: ,paspoort at dds.nl ...
Reverse transcription-polymerase chain reaction: …a laboratory technology known as reverse transcription-polymerase chain reaction (RT-PCR), a powerful tool used in research and in the diagnosis of diseases such as cancer.
Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, ...
A reverse transcription-polymerase chain reaction (RT-PCR) test is the only way to identify an infected COVID-19 person. Since it takes 24 to 36 hours to achieve results, the Indian Council for Medical Research (ICMR) recently introduced Chinese-made rapid tests kits which can produce results in 10 to 15 minutes. However, this exercise didnt prove successful because these test kits showed erratic results.. A lot of patients have complained that their first RT-PCR test was negative but later on, when the COVID-19 symptoms persisted and when they went for the second test, it came positive. They blame the diagnostic labs for these inconsistencies.. Diagnostic labs, which ICMR has approved for COVID-19 tests, say that the RT-PCR test can only detect 65 out of 100 positive patients because it has its own limitation. A whole gamut of factors decides the accuracy of the test.. This also explains why many people who have tested negative initially become positive later on.. Experts say that the presence ...
Mercedes Pérez-Ruiz, José-María Navarro-Marí, María-Fé Bautista-Marín, Irene Pedrosa-Corral, Sara Sanbonmatsu-Gámez, Ana G. Camacho, José Rojas, Jorge Ruiz-Ortiz, Javier Rodríguez-Granger, José Antonio Carrillo ...
Some gene transcripts represented by more than one probe set (each detected similar levels of expression) Cancer Genome Anatoy Project verified expression of many genes Q-RT-PCR -- some of 100 genes selected and measured for transcript levels using Q-RT-PCR --results in agreement with relative levels of microarray gene expression Immunohisotchemical analysis of tumor samples: established protein expression levels of select 100 correlated with mRNA levels from microarray Several genes identified have already been previously shown to be differentially expressed in metastatic prostate cancer ...
SOLIScript 1-step Multiplex Probe Kit is optimized for probe based one-step real-time quantitative RT-PCR assays. Learn about the product and order free samples here!
Lets set the record straight about DDRT-PCR! This technique recieves an amazing amount of bad press, which I find utterly astonishing given that has major advantages over other methods for identifying differentially expressed genes. It is an extremely powerful tool. I work in a research group of 6 scientists and we all routinely use DDRT-PCR for a variety of different purposes, using RNA extracted from a wide variety of material, ranging from cultured cells, specific tissues to whole embryos. We have been using the technique for the last couple of years with a great deal of success. My particular project is concerned with hunting for brain transcripts up- or down-regulated in developing mouse brain as a direct result of the targeted deletion of a specific gene. This is designed to give us clues as to the function of the encoded protein (the null has no phenotype). For this project I am using whole mouse brains from a variety of wt/null developmental timepoints, which DDRT-PCR allows me to ...
What could be the possible reasons for a RT-PCR experiment that was working fine - posted in PCR, RT-PCR and Real-Time PCR: What could be the possible reasons for a RT-PCR experiment that was working fine to stop working, if nothing was changed? I seem to see quite a few questions about this. And Ive been battling with this problem for quite a few months now.... Thank you! Sakumi
Methods:This study used 63 peripheral blood and bone marrow (PB/BM) samples from children with ALL. Immunophenotyping of PB and BM samples were performed using flow cytometry to illustrate the lineage. Moreover, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to amplify transcripts of leukemia-specific chromosome fusion gene ETV6/RUNX1 and to monitor the expression levels of the ETV6/RUNX1 in patients according to Van Dongen et al protocol ...
Two commercial PRRSV ELISA packages (IDEXX and Bionote) were evaluated for his or her sensitivity and specificity using 476 PRRS-positive serum examples collected from 7 animal challenge tests and 1,000 PRRS-negative sera. with the Chonbuk Country wide University Institutional GS-9350 Pet Care and Make use of Committee (acceptance amount: 2012-0025). 40 swine farms which have preserved PRRS-negative status within the last year were verified to be detrimental by real-time invert transcription-polymerase chain response (RT-PCR) and IDEXX PRRS 3XR Ab ELISA and had been selected for the analysis. Information concerning the primers and probe for the real-time RT-PCR is really as follows: ahead primer: TGTCAGATTCAGGGAGRATAAGTTAC; probe: TTTTGCACCACMGCCAGCCC; and invert primer: ATCARGCGCACAGTRTGATGC. RT-PCR was carried out using the AgPath-IDTM One-Step RT-PCR Package (Ambion, Austin, TX, U.S.A.) inside a 25 response quantity using 5 of extracted design template. PCR amplification included (a) GS-9350 ...
EurobioPlex SARS-CoV-2 is a CE marked test based on real-time reverse-transcription and amplification designed for qualitative determination of absence or…
Fig. 4 Tnnt temporal expression patterns in developing zebrafish embryos as detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Total RNA was isolated from different developmental stages, and primers specific for sMyHC, Tnnt1, Tnnt2, Tnnt3a, and Tnnt3b were used for RT-PCR. β-actin was used as an internal control. Numbers in the right panel indicate the molecular mass of RT-PCR products. ...
In this article explain the finer points of the delta-delta-CT method to calculate up-/down- regulations. The following text is a writeup of a course I gave at a local highschool.
Hepcidin, a peptide hormone that decreases intestinal iron absorption and macrophage iron release, is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. Endogenous stimulants of Hepcidin transcription include bone morphogenic protein 6 (BMP6) and interleukin-6 (IL-6) by effects on mothers against decapentaplegic homolog (Smad)4 or signal transducer and activator of transcription (Stat)3, respectively. We conducted a small-scale chemical screen in zebrafish embryos to identify small molecules that modulate hepcidin expression. We found that treatment with the isoflavone, genistein, from 28-52 hours postfertilization in zebrafish embryos enhanced Hepcidin transcript levels, as assessed by whole-mount in situ hybridization and quantitative real-time reverse-transcriptase polymerase chain reaction. Genisteins stimulatory effect was conserved in human hepatocytes: Genistein treatment of HepG2 cells increased both ...
Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Data from our previous
Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Data from our previous
Cancer stem cells are associated with metastatic potential, treatment resistance, and poor patient prognosis. Distant recurrence remains the major cause of mortality in rectal cancer patients with preoperative chemoradiotherapy (CRT). We investigated the role of three stem cell markers (CD133, OCT4, and SOX2) in rectal cancer and evaluated the association between these gene levels and clinical outcome in rectal cancer patients with preoperative CRT. Thirty-three patients with rectal cancer underwent preoperative CRT. Total RNAs of rectal cancer cells before and after CRT were isolated. Residual cancer cells after CRT were obtained from formalin-fixed paraffin-embedded (FFPE) specimens using microdissection. The expression levels of three stem cell genes were measured using real-time reverse-transcription polymerase chain reaction (RT-PCR). The association between these gene levels and radiation was evaluated using colon cancer cell lines. Immunohistochemical staining of these markers after CRT was also
Among diabetes-susceptibility genes in NOD mice, only Idd-1 has been clearly assigned: Idd-1 could be a gene complex composed of class II major histocompatibility complex (MHC) genes, I-A beta and I-E. Employing restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing, we revealed that ILI and CTS mice, which are nondiabetic but are derived from the same Jcl-ICR mice as NOD mice, share the same class II MHC genes with NOD mice suggesting that both ILI and CTS mice also possess susceptible Idd-1 genotype. This was supported by a breeding study. To compare the usage of T cell receptor (TCR) V beta genes in NOD mice with that in ILI mice, we employed quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) which revealed that TCR V beta usages of these mice were indistinguishable. RT-PCR method also revealed that the V beta transcript of T cells infiltrating into pancreas of NOD mice was not restricted but was rather diverse. Since NOD and ILI mice share the same
Five potential reference genes for RT-qPCR application, namely histone H3, beta-actin, GAPDH, ubiquitin and 18S rRNA, were evaluated for normalization of gene expression in four selected tissues (liver, kidney, thyroid and abdominal fat). Tissues were derived from fattening pigs exposed to different amounts and type of dietary iodine. Two software applications (geNorm and NormFinder) were used to evaluate the stability of the potential reference genes. All studied genes displayed high expression stability but different stability patterns between the investigated tissues. The results suggest GAPDH and 18S rRNA as reference genes applicable in all tissues investigated. Beta-actin and histone H3 are suitable reference genes for all tissues investigated except fat. In contrast, ubiquitin should be excluded from use as a reference gene in the porcine tissues analyzed due to variations in expression levels, despite the good expression stability.
A multiplex reverse transcription polymerase chain reaction (RT-PCR) protocol was evaluated for the simultaneous detection of Potato leaf roll virus (PLRV), Potato virus X (PVX) and Potato Virus Y (PVY). The multiplex RT-PCR detection of viruses was equally efficient whether RNAs were extracted using commercial kits or a sodium sulphite based nucleic acid extraction procedure. cDNAs were prepared either using a common primer (oligo dT) or specific antisense primer followed by specific primer pairs for PCR. The multiplex RT-PCR separation of strains of PVY was accomplished by using a competitive multiplex RT-PCR (with one antisense and two sense primers). The multi virus or multistrain detection approaches described here have potential application to other crop-virus combinations ...
Identification of potential reference genes. Potential reference genes are identified by (A) geNorm and (B) NormFinder. Low M-value or variability represents th
We report on the identification and genomic characterization of DEIN, a novel gene that exhibits a characteristic expression pattern in primary neuroblastoma. With a combined RACE and RT-PCR approach, the full-length sequences of the main transcript variants of this gene were characterized. According to SAGE, Northern blot, and quantitative real-time RT-PCR, variants A and B represent the major transcripts of DEIN in primary neuroblastoma. These isoforms are strongly expressed in stage IVS tumors, whereas lower expression levels were observed in stage IV tumors. In contrast, expression of transcript variant C did not differ in these stages, as determined by quantitative real-time RT-PCR, and seemed to be expressed at lower levels in neuroblastoma because it could not be detected by Northern blot hybridization and Ct values of quantitative real-time RT-PCR analysis were high (data not shown). In addition, low numbers of SAGE tag ATTTGTTACA corresponding to transcript D suggested that this isoform ...
To inform an example calculation, we use publicly published data for analytic specificity of the Quest Diagnostics reverse transcription polymerase chain reaction assay (likely 100%, minimum 95%, maximum 100%) and an informed but pessimistic assumption regarding the clinical sensitivity of the reverse transcription polymerase chain reaction assay (likely 90%, minimum 65%, maximum 99%). Estimation of population prevalence is challenging: the minimum in this scenario is based on a recent measurement of the prevalence of reverse transcription polymerase chain reaction positivity among asymptomatic individuals in Iceland (0.6%), while our maximum is based on a recently published estimate among asymptomatic parturients at a major academic center (13.8%).6,7 As is the case with nearly all measurements of disease prevalence, both of these estimates were measured in unique populations at specific points in time. We chose a most likely prevalence estimate of 1.0% based on preliminary, unpublished data ...
צעד אחד RT-PCR assay לגילוי וזיהוי genogroup של מבודד norovirus מ כיסאות ילדים, אשר מנצל primers ו בדיקות TaqMan ספציפיים מסגרת קריאה...
Reverse transcription polymerase chain reaction (RT‐PCR) expression patterns of the genes coding for Laccaria bicolor Zn finger protein and Ser/Threo protein
Quiescent and proliferating leukemia cells accumulate highly lethal DNA double-strand breaks that are repaired by 2 major mechanisms: BRCA-dependent homologous recombination and DNA-dependent protein kinase-mediated (DNA-PK-mediated) nonhomologous end-joining, whereas DNA repair pathways mediated by poly(ADP)ribose polymerase 1 (PARP1) serve as backups. Here we have designed a personalized medicine approach called gene expression and mutation analysis (GEMA) to identify BRCA- and DNA-PK-deficient leukemias either directly, using reverse transcription-quantitative PCR, microarrays, and flow cytometry, or indirectly, by the presence of oncogenes such as BCR-ABL1. DNA-PK-deficient quiescent leukemia cells and BRCA/DNA-PK-deficient proliferating leukemia cells were sensitive to PARP1 inhibitors that were administered alone or in combination with current antileukemic drugs. In conclusion, GEMA-guided targeting of PARP1 resulted in dual cellular synthetic lethality in quiescent and proliferating ...
Tissue RNA PrepMate™ uses a modified guanidium salt-lysis method to optimally extract total RNA from tissue. The amount of extracted total RNA can differ according to the tissue or cell type, so the starting amount should be adjusted to your needs. The extracted viral RNA can be used as a template of the RT-PCR or quantitative real time RT-PCR and cDNA synthesis, cDNA library construction, Microarray ...
Pregnancy in mammals featuring hemochorial placentation introduces a major conflict with the mothers immune system, which is dedicated to repelling invaders bearing foreign DNA and RNA. Numerous and highly sophisticated ...
This is an historical archive of the activities of the MRC Anatomical Neuropharmacology Unit (MRC ANU) that operated at the University of Oxford from 1985 until March 2015. The MRC ANU established a reputation for world-leading research on the brain, for training new generations of scientists, and for engaging the general public in neuroscience. The successes of the MRC ANU are now built upon at the MRC Brain Network Dynamics Unit at the University of Oxford.. ...
Naina Mishra. Tribune News Service. Chandigarh, July 29. The UT Administration is set to introduce the rapid antigen test for the first time for the diagnosis of Covid-19.. According to sources, the UT will procure around 2,000 kits in the coming days. A tender has been floated on the GeM portal.. The UT will use the antigen test for asymptomatic patients only. Those who test positive will be considered Covid patients. As per the ICMR advisory, symptomatic patients with negative antigen results have to go in for the RT-PCR test for confirmation. However, no RT-PCR test confirmation is required for asymptomatic individuals testing negative in the antigen test.. However, the antigen test has been widely condemned for its low sensitivity as compared to the RT-PCR test. In its advisory, the ICMR said it had evaluated the antigen kits performance in two labs and had found its sensitivity to be 50.6 per cent and 84 per cent.. Recently, the Delhi High Court had asked the Delhi Government why it was ...
Recently we have reported that intermediate-frequency magnetic field (IF-MF) exposure transiently altered the mRNA expression levels of memory function-related genes in the hippocampi of adult male mice. However, the effects of IF-MF exposure during brain development on neurological biomarkers have not yet been clarified. In the present study, we investigated the effect of IF-MF exposure during development on neurological and immunological markers in the mouse hippocampus in 3- and 7-week-old male mice. Pregnant C57BL/6J mice were exposed to IF-MF (21 kHz, 3.8 mT) for one hour per day from organogenesis period day 7 to 17. At adolescence, some IF-MF-exposed mice were further divided into exposure, recovery, and sham-exposure groups. The adolescent-exposure groups were exposed again to IF-MF from postnatal day 27 to 48. The expression of mRNA in the hippocampi was examined using a real-time RT-PCR method, and microglia activation was examined by immunohistochemical analysis. The expression levels of NR1
Using the laboratory RT-PCR method, we detect the presence of SARS-CoV-2 virus in the body. A negative result can be used to end the quarantine and is required for travelling to and from abroad. The test will take place in a general practitioner's office in Prague. You will usually find out the result the next day after collection. The result will be sent to you by e-mail.
Molecular Cytogenetics and Multiplex Reverse-Transcriptase Polymerase Chain Reaction for Risk Stratification in Acute Myeloid Leukemia ...
Background: Current methodology of gene expression analysis limits the possibilities of comparison between cells/tissues of organs in which cell size and/or number changes as a consequence of the study (e.g. starvation). A method relating the abundance of specific mRNA copies per cell may allow direct comparison or different organs and/or changing physiological conditions. Methods: With a number of selected genes, we analysed the relationship of the number of bases and the fluorescence recorded at a present level using cDNA standards. A lineal relationship was found between the final number of bases and the length of the transcript. The constants of this equation and those of the relationship between fluorescence and number of bases in cDNA were determined and a general equation linking the length of the transcript and the initial number of copies of mRNA was deduced for a given pre-established fluorescence setting. This allowed the calculation of the concentration of the corresponding mRNAs per ...
There are currently no tests available over the counter or at a doctors office that can quickly detect and distinguish between the H7N9 virus and other flu viruses. However, a more sophisticated test that specifically detects H7N9 virus has been developed by CDC for use by qualified public health laboratories in the United States and internationally. This test involves collecting a respiratory tract (i.e., nose, throat, lung) sample from a sick patient. The sample is then sent to a public health laboratory where a procedure known as rRT-PCR (real-time reverse transcriptase polymerase chain reaction) is conducted. rRT-PCR is very accurate and sensitive at detecting flu viruses. This procedure typically provides results within 4 hours; however, the time involved in processing and reporting results may vary depending on the laboratory ...
Main focus of GENE QUANTIFICATION web page is to describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time RT-PCR and competitive RT-PCR. It illustrates the usefulness of absolute and relative quantification assays in real-time PCR and real-time RT-PCR.
In order to identify biomarkers to predict the progression of Type 1 Diabetic Kidney Disease (DKD), we profiled the transcriptome (the set of RNA molecules produced) of leukocytes from 33 type 1 diabetic patients at the time of their enrollment in the study. Patients were followed for a minimum of five years and data were collected including information about the progression of DKD. We will identify genes that correlate to various outcomes with a focus on the decline of kidney function. To validate the expression, we will use a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR,) a laboratory method that is used to quantify RNA. Validated genes will be further studied in the future with more patients.. ...
ELISA and real-time qRT-PCR analyses demonstrated an increase of VEGF protein and mRNA levels in the inner ear after local EP2 or EP4 agonist application, indicating that the stimulation of EP2 and/or EP4 activates VEGF production in the inner ear ...
Cancer cells. (A) Relative expression levels of Nox1, 2, 3, 4, and 5 mRNAs in A549 cells were determined by real-time RT-PCR and are presented as mean delta Ct
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Test with Confidence. Test with ZEUS. Clinical Performance Studies Four separate cohorts of clinically characterized specimens were tested: COVID-19 RT-PCR Positive Patient Specimens (n=35). The date between EUA-approved PCR test result and specimen draw was 3 to 37 days, with an average of 15.97 days and a median of 14 days.COVID-19 RT-PCR Negative Patient Specimens…
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed ...
Video created by Exploratorium for the course Tinkering Fundamentals: Motion and Mechanisms. A Chain Reaction machine is a deceptively simple concept, but one that allows for an incredibly complex and deep investigation into something we ...
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Shop a large selection of Invitrogen™ CellsDirect™ One-Step qRT-PCR Kit products and learn more about Invitrogen™ CellsDirect™ One-Step qRT-PCR
A correlative immunohistochemical and reverse-transcriptase polymerase chain reaction analysis". Virchows Arch. 440 (5): 461-75 ...
A correlative immunohistochemical and reverse-transcriptase polymerase chain reaction analysis". Virchows Archiv. 440 (5): 461- ...
Grillo M, Margolis FL (September 1990). "Use of reverse transcriptase polymerase chain reaction to monitor expression of ... Becker-André M, Hahlbrock K (November 1989). "Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel ... RNA is first copied as complementary DNA (cDNA) by a reverse transcriptase enzyme before the resultant cDNA is sequenced. The ... and later Reverse Transcriptase quantitative PCR (RT-qPCR) methods, but these methods are laborious and can only capture a tiny ...
Biswal M, Ratho R, Mishra B (September 2007). "Usefulness of reverse transcriptase-polymerase chain reaction for detection of ... 10: PCR technology for lyssavirus diagnosis". In Clewley, J.P. The Polymerase Chain Reaction (PCR) for Human Viral Diagnosis. ... the viral polymerase will begin to synthesize new negative strands of RNA from the template of the positive strand RNA. These ... and the viral RNA polymerase (L). Once within a muscle or nerve cell, the virus undergoes replication. The trimeric spikes on ...
... in gynecologic malignancies and malignant effusions detected by nested reverse transcriptase-polymerase chain reaction". ...
... the altered activation will be examined by reverse-transcriptase-polymerase chain reaction (RTPCR) analysis of induced genes. ...
Feline coronavirus quantitative reverse transcriptase polymerase chain reaction on effusion samples in cats with and without ... Comparison of real-time reverse transcriptase polymerase chain reaction of peripheral blood mononuclear cells, serum and cell- ... Sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction detecting feline coronavirus ... by reverse-transcriptase polymerase chain reaction (RT-PCR) is diagnostic of effusive FIP. However, that does require that a ...
Reverse transcriptase polymerase chain reaction (RT-PCR) methods have been developed for detection of TSV and are very ... The IQ2000TM TSV detection system, a RT-PCR method, is said to have a detection limit of 10 copies per reaction. RNA-based ... These very high rates might be due to the lack of proofreading function of the RNA-dependent RNA polymerase and have resulted ...
... prognostic value of detection of circulating colorectal cancer cells using CGM2 reverse transcriptase-polymerase chain reaction ...
The genome is detectable by reverse transcriptase-polymerase chain reaction in the head, thorax, abdomen and wings of infected ... The genome structure is 5'UTR-L-VP2-(VP4)-VP1-VP3-RNA helicase-(VPg)-3C protease-RNA dependent RNA polymerase-3'UTR The ... a chymotrypsin-like 3C protease and an RNA-dependent RNA polymerase. VP1 is encoded between codons 486 to 880 and VP3 lies ...
ISBN 978-0-470-84475-5. Callahan JD, et al., Use of a portable real-time reverse transcriptase-polymerase chain reaction assay ... Quantitative PCR utilizes polymerase chain reaction chemistry to amplify viral DNA or RNA to produce high enough concentrations ... "Taqman Technology and Real-Time Polymerase Chain Reaction". In Crocker, J.; Murray, P.G. Molecular Biology in Cellular ... SYBR Green dye binds to all double-stranded DNA produced during the reaction. While SYBR Green is easy to use, its lack of ...
The role of reverse transcriptase polymerase chain reaction assay for prostate specific antigen in the selection of patients ... "Enhanced Reverse Transcriptase Polymerase Chain Reaction for Prostate-specific Antigen Combined With Needle Biopsy Results: A ... The detection of renal Carcinoma cells in the peripheral blood with an enhanced reverse transcriptase-polymerase chain reaction ...
These tags can be appended to gene specific primers for reverse transcriptase polymerase chain reaction (RT-PCR) experiments, ... In order to overcome genomic DNA contamination in transcriptional studies, reverse template-specific polymerase chain reaction ... of using tags whose sequences are not found in the genome further improves reverse specific polymerase chain reaction ... Shuldiner, Alan R.; Ajay Nirula; Jesse Roth (1990). "RNA template-specific polymerase chain reaction (RS-PCR): a novel strategy ...
Ikeguchi M, Hirooka Y, Kaibara N (Sep 2003). "Quantitative reverse transcriptase polymerase chain reaction analysis for KiSS-1 ...
2006). "A validated quantitative assay to detect occult micrometastases by reverse transcriptase-polymerase chain reaction of ... 2009). "GUCY2C reverse transcriptase PCR to stage pN0 colorectal cancer patients". Expert Rev. Mol. Diagn. 9 (8): 777-85. doi: ...
... and Application for Localization in Various Human Tissues by Real-Time Reverse Transcriptase-Polymerase Chain Reaction". Drug ... glucuronides is known to occur to nucleophilic sites on serum albumin via transacylation reactions, for example. Phenols, ...
... and Application for Localization in Various Human Tissues by Real-Time Reverse Transcriptase-Polymerase Chain Reaction". Drug ... Al-Zoughool M., Talaska, G. (2006). "4-Aminobiphenyl N-glucuronidation by liver microsomes: optimization of the reaction ...
The identification of HMPV has predominantly relied on reverse-transcriptase polymerase chain reaction (RT-PCR) technology to ...
"Detection of human 11 beta-hydroxysteroid dehydrogenase isoforms using reverse-transcriptase-polymerase chain reaction and ... In addition, the encoded protein can catalyze the reverse reaction, the conversion of cortisone to cortisol. Too much cortisol ...
... a reverse transcriptase-polymerase chain reaction (RT-PCR) study, Eastwood et al., Molecular Brain Research Vol44, Iss1, ... reversing the effects of endocytosis and LTD. when appropriate.[66] Nevertheless, the highlighted calcium-dependent, dynamin- ... of excitatory neurotransmission by decanoic acid in the brain contributes to the anticonvulsant effect of the medium-chain ...
Reverse transcriptase polymerase chain reaction (RT-PCR) has become a powerful and effective method for detection of potato ... Different types of reverse transcriptase polymerases are available to suit different needs and reaction conditions. Reverse ... It could also be that the reverse transcriptase polymerase and DNA polymerase is one and the same enzyme and that the enzyme ... the RNA of the virus must first be transcribed to DNA by means of a reverse transcriptase polymerase. This polymerase ...
... reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ-RT-PCR for the detection of canine distemper virus RNA in ... and other evidence suggests an intrinsic hyperresponsive reaction to vitamin D and RANK ligand is the cause.[citation needed] ...
... a reverse transcriptase-polymerase chain reaction (RT-PCR) study, Eastwood et al., Molecular Brain Research Vol44, Iss1, ... For the latter, PICK1 and PKC can displace GRIP1 to return AMPARs to the surface, reversing the effects of endocytosis and LTD ... of excitatory neurotransmission by decanoic acid in the brain contributes to the anticonvulsant effect of the medium-chain ...
Splice modification can be conveniently assayed by reverse-transcriptase polymerase chain reaction (RT-PCR) and is seen as a ... Morpholinos are used as research tools for reverse genetics by knocking down gene function. This article discusses only the ...
... other interstitial lung disease during clinical studies applying quantitative reverse transcriptase-polymerase chain reaction ( ...
... and reverse transcriptase activity (using reverse transcriptase polymerase chain reaction techniques) from neoplastic tissue. ... amino acid identity in the pol region of reverse transcriptase. This finding suggests that they are different strains of the ...
... reverse-transcriptase polymerase chain reaction, and nucleic acid microarrays for specialized studies of disease in tissues and ...
... reverse transcriptase polymerase chain reaction (sometimes real-time polymerase chain reaction). Clarified with "quantitative ... "reverse transcriptase PCR" where appropriate. RTS - see entry RTT - (i) Round Trip Timing RTV - (i) Rapid Terrain Visualization ... Reverse Osmosis Water Purification Unit ("roe-pew") ROY G BIV - colors of the prism; Red, Orange, Yellow, Green, Blue, Indigo, ... Reverse Polish Lisp rpm - (s) revolutions per minute RPM (i) Records, Promotion, Music (expansion of the title of a former ...
Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR), which is based on the analysis of RNA rather than DNA, ... A real-time polymerase chain reaction (Real-Time PCR), also known as quantitative polymerase chain reaction (qPCR), is a ... The polymerase chain reaction (PCR) is a common method for amplifying DNA; for RNA-based PCR the RNA sample is first reverse- ... The acronym "RT-PCR" commonly denotes reverse transcription polymerase chain reaction and not real-time PCR, but not all ...
... isothermal alternative to the polymerase chain reaction (PCR). By adding a reverse transcriptase enzyme to an RPA reaction it ... although users can simply supplement other TwistAmp reactions with a reverse transcriptase to produce the same effect. As with ... If a reverse transcriptase that works at 37-42 °C is added then RNA can be reverse transcribed and the cDNA produced amplified ... "A Portable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of Foot-and-Mouth Disease Virus ...
... polymerase - Polymerase chain reaction (PCR) - polyneuritis - polypeptide - polyvalent vaccine - post-exposure prophylaxis (PEP ... reverse transcriptase - ribonucleic acid (RNA) - ribosome - RNA - route of administration - RT-PCR - RTI - Ryan White C.A.R.E. ... non-nucleoside reverse transcriptase inhibitors (NNRTI) - non-steroidal anti-inflammatory drugs (NSAID) - NRTI - nucleic acid ... nucleoside reverse transcriptase inhibitors (NRTI) - nucleotide - nucleotide analogs - nucleus - null cell ...
A polymerase chain reaction (PCR) then coats each bead with clonal copies of the DNA molecule followed by immobilization for ... using reverse transcriptase-a DNA polymerase that synthesizes a complementary DNA based on existing strands of RNA in a PCR- ... "Use of automated sequencing of polymerase chain reaction-generated amplicons to identify three types of cholera toxin subunit B ... Chain-termination methods[edit]. Main article: Sanger sequencing. The chain-termination method developed by Frederick Sanger ...
Technologies based upon the polymerase chain reaction (PCR) method will become nearly ubiquitous gold standards of diagnostics ... For example, humans can make neither RNA replicases nor reverse transcriptase, and the presence of these enzymes are ... There is a general chain of events that applies to infections.[22] The chain of events involves several steps-which include the ... An infection is the invasion of an organism's body tissues by disease-causing agents, their multiplication, and the reaction of ...
This is overcome by polymerase chain reaction (PCR) amplification. Digital representationEdit. ... Instead, it is copied to a DNA by reverse transcriptase, and this DNA is then sequenced. ... Nucleic acids consist of a chain of linked units called nucleotides. Each nucleotide consists of three subunits: a phosphate ... Current sequencing methods rely on the discriminatory ability of DNA polymerases, and therefore can only distinguish four bases ...
... and using a lower number of ditag polymerase chain reactions (PCR) to obtain a complete cDNA library.[11] ... The mRNA of an input sample (e.g. a tumour) is isolated and a reverse transcriptase and biotinylated primers are used to ... and reverse transcriptase is used to copy the mRNA into stable double-stranded-cDNA (ds-cDNA; blue). In SAGE, the ds-cDNA is ... The cleaved cDNA tags are then repaired with DNA polymerase to produce blunt end cDNA fragments. ...
Zhou C, Liu J (March 2003). "Inhibition of human telomerase reverse transcriptase gene expression by BRCA1 in human ovarian ... RNA polymerase II transcription coactivator activity. • RNA polymerase binding. • identical protein binding. ... Wu-Baer F, Lagrazon K, Yuan W, Baer R (September 2003). "The BRCA1/BARD1 heterodimer assembles polyubiquitin chains through an ... "Control of biochemical reactions through supramolecular RING domain self-assembly". Proc. Natl. Acad. Sci. U.S.A. 99 (24): ...
... reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ-RT-PCR for the detection of canine distemper virus RNA in ... and other evidence suggests an intrinsic hyperresponsive reaction to vitamin D and RANK ligand is the cause.[citation needed] ...
HIV reverse transcriptase is an enzyme for AIDS virus replication.[126] Telomerase is an unusual polymerase because it contains ... such as restriction digests and the polymerase chain reaction. Modern biology and biochemistry make intensive use of these ... Polymerases. Polymerases are enzymes that synthesize polynucleotide chains from nucleoside triphosphates. The sequence of their ... They include reverse transcriptase, which is a viral enzyme involved in the infection of cells by retroviruses, and telomerase ...
... was designed with a chloroindole part in wing I to expand interactions with the side chain of conserved W229 of the polymerase ... shown that the NNRTIs have other mechanisms of action and interfere with various steps in the reverse transcriptase reaction. ... nucleoside/nucleotide reverse-transcriptase inhibitors (NRTIs/NtRTIs) and non-nucleoside reverse-transcriptase inhibitors ( ... Reverse transcriptase (RT) is an enzyme that controls the replication of the genetic material of HIV and other retroviruses. ...
Polymerase Chain Reaction/PCR) দ্বাৰা অসংখ্য একে ধৰণৰ আকাংক্ষিত DNA প্ৰস্তুত কৰিবলৈ টেক্ DNA পলিমেৰেজ উসেচক ব্যৱহাৰ কৰা হয়। ( ... Reverse transcriptase): mRNA ক সাচ দণ্ড (Template strand) হিচাপে ব্যৱহাৰ কৰি একদণ্ডযুক্ত পৰিপূৰক DNA (cDNA) সৃষ্টি হওঁতে এই ... Polymerase Chain Reaction/PCR)। (a) জিন ক্লনিং (Gene Cloning): জীৱ কোষৰ ভিতৰত (in vivo) ৰিকম্‌বিনেণ্ট DNA প্ৰস্তুত কৰি উপযুক্ত ... Polymerase Chain Reaction): পলিমেৰেজ উৎসেচকৰ সহায়ত দুই ধৰণৰ অলিগ'নিউক্লিয়'টাইড প্ৰাইমাৰ (Oligonucleotide primers), ...
Reverse Transcription PCR (RT-PCR): for amplifying DNA from RNA. Reverse transcriptase reverse transcribes RNA into cDNA, which ... The Polymerase Chain Reaction".. *^ "Determining Annealing Temperatures for Polymerase Chain Reaction". The American Biology ... In vitro Amplification of DNA by the Polymerase Chain Reaction *^ "Polymerase Chain Reaction (PCR)". National Center for ... "An Overview of Nanoparticle-Assisted Polymerase Chain Reaction Technology". An Overview of Nanoparticle‐Assisted Polymerase ...
Zhou, Chenyi; Liu Jinsong (2003). «Inhibition of human telomerase reverse transcriptase gene expression by BRCA1 in human ... RNA polymerase II transcription coactivator activity. • RNA polymerase binding. • identical protein binding. ... Kentsis, Alex; Gordon Ronald E, Borden Katherine L B (2002). «Control of biochemical reactions through supramolecular RING ... Wu-Baer, Foon; Lagrazon Karen, Yuan Wei, Baer Richard (2003). «The BRCA1/BARD1 heterodimer assembles polyubiquitin chains ...
Technologies based upon the polymerase chain reaction (PCR) method will become nearly ubiquitous gold standards of diagnostics ... For example, humans can make neither RNA replicases nor reverse transcriptase, and the presence of these enzymes are ... There is a general chain of events that applies to infections.[12] The chain of events involves several steps-which include the ... This binding then sets off a chain of events that can be visibly obvious in various ways, dependent upon the test. For example ...
Polymerase. DNA polymerase. DNA-directed DNA polymerase. I. II. III. IV. V. RNA-directed DNA polymerase. Reverse transcriptase ... Mullis KB (April 1990). "The unusual origin of the polymerase chain reaction". Sci. Am. 262 (4): 56-61, 64-5. doi:10.1038/ ... RNA polymerase/DNA-directed RNA polymerase. RNA polymerase I. II. III. IV. V. Primase. RNA-dependent RNA polymerase. PNPase. ... It is frequently used in the polymerase chain reaction (PCR), a method for greatly amplifying the quantity of short segments of ...
RNA-directed DNA polymerase. Reverse transcriptase Telomerase. RNA nucleotidyltransferase. Template-directed. RNA polymerase I ... This CK enzyme reaction is reversible and thus ATP can be generated from PCr and ADP. ... Branched-chain amino acid aminotransferase. *Branched-chain alpha-keto acid dehydrogenase complex ... Polymerase. DNA polymerase. DNA-directed DNA polymerase. I/A γ. θ. ν. T7. Taq. II/B α. δ. ε. ζ. Pfu. III/C. IV/X β. λ. μ. TDT. ...
Vmax will decrease due to the inability for the reaction to proceed as efficiently, but Km will remain the same as the actual ... This is a potent enzyme inhibitor, in this case preventing the RNA polymerase II enzyme from transcribing DNA.[53] The algal ... These electrophilic groups react with amino acid side chains to form covalent adducts. The residues modified are those with ... Irreversible inhibitors usually covalently modify an enzyme, and inhibition can therefore not be reversed. Irreversible ...
One lineage led to the modern DNA Polymerases and reverse transcriptases, as well as to a few single-subunit RNA polymerases ( ... The second Mg2+ will hold on to the pyrophosphate of the NTP.[17] The overall reaction equation is: (NMP)n + NTP → (NMP)n+1 + ... It then produces an RNA chain, which is complementary to the template DNA strand. The process of adding nucleotides to the RNA ... Polymerase. DNA polymerase. DNA-directed DNA polymerase. I/A γ. θ. ν. T7. Taq. II/B α. δ. ε. ζ. Pfu. III/C. IV/X β. λ. μ. TDT. ...
... a polymerase chain reaction (PCR), western blot or, less commonly, an immunofluorescence assay (IFA)). Only specimens that are ... Bonhoeffer et al.[80] suggested that template switching by reverse transcriptase acts as a repair process to deal with breaks ... The reverse transcriptase also has ribonuclease activity that degrades the viral RNA during the synthesis of cDNA, as well as ... Shortly after the viral capsid enters the cell, an enzyme called reverse transcriptase liberates the positive-sense single- ...
This principle is the basis of commonly performed laboratory techniques such as the polymerase chain reaction, PCR.[1] ... cDNA libraries are constructed from mRNA using RNA-dependent DNA polymerase reverse transcriptase (RT), which transcribes an ... Reverse-complement tool page with documented IUPAC code conversion, source code available. ... much like looking in the mirror and seeing the reverse of things. This complementary base pairing allows cells to copy ...
Polymerase. DNA polymerase. DNA-directed DNA polymerase. I. II. III. IV. V. RNA-directed DNA polymerase. Reverse transcriptase ... The chemical reaction performed by these enzymes can be written as ATP + a protein ⇌. {\displaystyle \rightleftharpoons }. ADP ... Myosin-heavy-chain kinase (EC *Aurora kinase *Aurora A kinase ... RNA polymerase/DNA-directed RNA polymerase. RNA polymerase I. II. III. IV. V. Primase. RNA-dependent RNA polymerase. PNPase. ...
Non-nucleoside reverse-transcriptase inhibitors. *NS5A inhibitors. *Nucleoside and nucleotide reverse-transcriptase inhibitors ... Myosin-heavy-chain kinase (EC *Aurora kinase *Aurora A kinase ... That reaction breaks the binding of BAD to BCL-XL and BCL2, a mitochondrial death inhibitors, resulting in inactivation of BAD[ ... DNA polymerase inhibitor (Cytarabine#). *Ribonucleotide reductase inhibitor (Gemcitabine#). *Hypomethylating agent (Azacitidine ...
Kary Mullis demonstrates the polymerase chain reaction, a method for amplifying specific bits of DNA (1983). ... Demonstration of the role of reverse transcriptases in tumor viruses, independently by Howard Temin and David Baltimore, 1970. ... Enrico Fermi and his team in Rome achieved a nuclear reaction (1934, although the results were not understood until 1938, when ... Antoine Lavoisier determines that chemical reactions in a closed container do not alter total mass. From these observations he ...
2014). "Reverse transcriptase droplet digital PCR shows high resilience to PCR inhibitors from plant, soil and water samples". ... The polymerase chain reaction method is used to quantify nucleic acids by amplifying a nucleic acid molecule with the enzyme ... Digital polymerase chain reaction (digital PCR, DigitalPCR, dPCR, or dePCR) is a biotechnological refinement of conventional ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information, U.S. National Library of Medicine. Duewer, ...
Kary Mullis demonstrates the polymerase chain reaction, a method for amplifying specific bits of DNA (1983). The experiments of ... Demonstration of the role of reverse transcriptases in tumor viruses, independently by Howard Temin and David Baltimore, 1970. ... Frederick Sanger demonstrates the dideoxy- or chain termination method for determining DNA sequences (1975). ... Enrico Fermi and his team in Rome achieved a nuclear reaction (1934, although the results were not understood until 1938, when ...
A polymerase chain reaction is a form of enzymatic DNA synthesis in the laboratory, using cycles of repeated heating and ... using reverse transcriptase enzymes. In retroviruses, viral RNA is inserted into a host cell nucleus. There, a viral reverse ... polymerase chain reaction - enzymatic DNA synthesis (in vitro DNA amplification) or gene synthesis - physically creating ... The technique couples a reverse transcription reaction with PCR-based amplification, as an RNA sequence acts as a template for ...
One widely adopted WGA techniques is called degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). This method ... The first step in quantifying the transcriptome is to convert RNA to cDNA using reverse transcriptase so that the contents of ... usually Φ29 DNA polymerase, to accomplish the amplification of larger fragments and greater genome coverage than DOP-PCR. ...
Ricchetti M, Buc H (February 1993). "E. coli DNA polymerase I as a reverse transcriptase". The EMBO Journal. 12 (2): 387-96. ... In fact, the Klenow fragment was used during the first protocols of polymerase chain reaction (PCR) amplification until Thermus ... which retains only the DNA polymerase and proofreading activities. DNA polymerase II DNA polymerase III DNA polymerase V Lehman ... Studies of polymerase I have confirmed that different dNTPs can bind to the same active site on polymerase I. Polymerase I is ...
  • A comparison of in situ hybridisation, reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ-RT-PCR for the detection of canine dist. (nih.gov)
  • A comparison of in situ hybridisation, reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ-RT-PCR for the detection of canine distemper virus RNA in Paget's disease. (nih.gov)
  • Reverse-transcriptase-polymerase chain reaction (RT-PCR) has been applied successfully for detection of canine distemper virus (Agnihotri et al. (thefreelibrary.com)
  • The detection of prostate-specific antigen (PSA) mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR) in the bloodstream of prostate cancer patients has been hypothesized as a prognostic marker, however little data are available concerning the association between this molecular marker and other laboratory values of potential importance. (elsevier.com)
  • Detection of circulating malignant cells (CMCs) through a reverse transcriptase-polymerase chain reaction (RT-PCR) assay seems to be a demonstration of systemic disease. (biomedcentral.com)
  • Optimization of the reverse transcriptase polymerase chain reaction for the detection of circulating prostate cells. (openrepository.com)
  • Reverse transcription-polymerase chain reaction detection of prostate-specific antigen, prostate-specific membrane antigen, and prostate stem cell antigen in one milliliter of peripheral blood: value for the staging of prostate cancer. (openrepository.com)
  • Detection of circulating prostate cells by reverse transcriptase-polymerase chain reaction of human glandular kallikrein (hK2) and prostate-specific antigen (PSA) messages. (openrepository.com)
  • To improve on existing methods of detection, we have developed a reverse-transcriptase polymerase chain reaction (RT-PCR) assay for keratin 19 (K19) transcripts to identify mammary carcinoma cells in the peripheral blood and bone marrow of patients with breast cancer. (umn.edu)
  • Reverse transcriptase/polymerase chain reaction detection of cytokeratin-19 mRNA in bone marrow and blood of breast cancer patients. (wizdom.ai)
  • 1990. Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. (koreascience.or.kr)
  • In order to establish an accurate, ready-to-use assay for simultaneous detection of Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus (WEEV), we developed one duplex TaqMan real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay, which can be used in human and vector surveillance. (biomedcentral.com)
  • Bowers, RM, Lapatra, SE & Dhar, AK 2008, ' Detection and quantitation of infectious pancreatic necrosis virus by real-time reverse transcriptase-polymerase chain reaction using lethal and non-lethal tissue sampling ', Journal of Virological Methods , vol. 147, no. 2, pp. 226-234. (elsevier.com)
  • We show here a sensitive and reproducible real- time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. (bvsalud.org)
  • The aim of this study was to evaluate the Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) and Heminested RT-PCR (hnRT-PCR) techniques for the detection of rabies virus genome in brain samples stored at -20°C (average freezers) for distinct periods and after decomposition for 72 hours in room temperature. (biomedcentral.com)
  • An analysis of the transcript level of the 10 members of the family has been performed using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. (plantphysiol.org)
  • Real-time quantitative reverse transcriptase polymerase chain reaction. (elsevier.com)
  • The real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) has become the method of choice for the quantification of specific mRNAs. (elsevier.com)
  • Fingerprint Dive into the research topics of 'Real-time quantitative reverse transcriptase polymerase chain reaction. (elsevier.com)
  • A reverse transcriptase-polymerase chain reaction assay was used to detect measles virus RNA in peripheral blood mononuclear cells, urine, and nasopharyngeal specimens from Zambian children during hospitalization and approximately 1-2 months after discharge. (lshtm.ac.uk)
  • The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. (koreascience.or.kr)
  • Using a series of dilutions of transcripts containing target genes as template, we showed that the sensitivity of the assay reached 1 copy/reaction for EEEV and WEEV, and the performance was linear within the range of at least 10 6 transcript copies. (biomedcentral.com)
  • A PCR solution is made similarly to a TaqMan assay, which consists of template DNA (or RNA), fluorescence-quencher probes, primers, and a PCR master mix, which contains DNA polymerase, dNTPs, MgCl2, and reaction buffers at optimal concentrations. (wikipedia.org)
  • Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer. (openrepository.com)
  • Positive prostate-specific antigen circulating cells detected by reverse transcriptase-polymerase chain reaction does not imply the presence of prostatic micrometastases. (openrepository.com)
  • Consequently, this work was directed toward development of a reverse-transcriptase polymerase chain reaction (RT-PCR)-based in vitro DNA amplification method, which specifically detects only viable cells. (nih.gov)
  • Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to measure the relative expression levels of mRNAs of the SOD1 , TGF-β1 , and DUSP1 genes. (molvis.org)
  • Candidate genes were further analyzed with reverse transcriptase polymerase chain reaction (RT-PCR) for their retinal specificity. (molvis.org)
  • Among the 25 clones that showed at least a 2-fold difference in expression was the gene of perilipin, upregulated in ruptured plaques, and the genes coding for fibronectin and immunoglobulin λ chain, which were downregulated in ruptured plaques. (ahajournals.org)
  • Design and Methods Expression of c-Kit isoforms was investigated by reverse transcriptase polymerase chain reaction in fresh plasma cells from patients and cell lines. (haematologica.org)
  • Cytokine-induced gene expression of several NF-κB targets, namely inducible nitric oxide synthase, Fas, and manganese superoxide dismutase, was prevented by AdIκB (SA)2 , as established by reverse transcriptase-polymerase chain reaction, protein blot, and measurement of nitrite in the medium. (diabetesjournals.org)
  • Non-muscle myosin 2 (NM II) is a hexameric protein composed of two dimeric heavy chains (NMHCs), a pair of regulatory light chains (RLCs) and a pair of essential light chains. (biologists.org)
  • METHODS Analysis of SwissProt protein and EMBL/GenBank nucleotide sequence banks, protein sequence alignment, reverse transcriptase-polymerase chain reaction and nucleotide sequencing were used. (bmj.com)
  • Quantification of HPV-16 E6-E7 transcription in cervical intraepithelial neoplasia by reverse transcriptase polymerase chain reaction. (nih.gov)
  • A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. (sickkids.ca)
  • Reverse-transcription/limiting-dilution polymerase chain reaction analysis showed that in response to IFN/TNF/IL-1, mRNA abundance of GTPCH and PTPS was increased ≈64-fold and 10-fold, respectively. (ahajournals.org)
  • a laboratory technology known as reverse transcription-polymerase chain reaction (RT-PCR), a powerful tool used in research and in the diagnosis of diseases such as cancer. (britannica.com)
  • Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome. (bmj.com)
  • 2014). This study is an attempt to compare polymerase chain reaction based molecular tests with commercially available IC based test for diagnosis of Canine distemper by testing rectal swabs and serum samples respectively from dogs suffering from gastroenteritis and suspected for Canine distemper. (thefreelibrary.com)
  • Recently, our group has synthesized a new biocompatible nanocarrier, poly(ester amine) (degradable polyethylenimine-alt-poly[ethylene glycol] copolymer), by reaction of low-molecular-weight polyethyleneimine (PEI) with polyethyleneglycol (PEG) diacrylate (number average molecular weight [M^sub n^] = 258) as a cross-linker. (redorbit.com)
  • A subsequent molecular analysis performed using reverse transcriptase-polymerase chain reaction with RNA extracted from paraffin-embedded tissue, revealed SYT/SSX1 fusion gene which confirmed the diagnosis of synovial sarcoma. (who.int)
  • The aim of this study was to evaluate sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) specifically designed to detect FCoV spike gene mutations at two nucleotide positions. (physiciansweekly.com)
  • Subject 'reverse transcriptase polymerase chain reaction' Remove constraint Subject: 'reverse transcriptase polymerase chain reaction' Journal 3 Biotech Remove constraint Journal: 3 Biotech Subject gene expression Remove constraint Subject: gene expression Subject Fusarium oxysporum f. sp. (usda.gov)
  • Method Serial analysis of gene expression (SAGE) and reverse transcriptase-polymerase chain reaction were used to identify RNA transcripts which are differentially expressed in the frontal cortex of brains obtained postmortem from individuals with bipolar disorder compared with other psychiatric and control conditions. (rcpsych.org)
  • PCR reactions for VEGF gene as well as internal control (β-actin) was performed 3 times using 2 -ΔΔCT (Livac) method for all specimens. (scitechnol.com)
  • Gene rearrangement of the Ig heavy chain indicated monoclonality or oligoclonality in all lymphomas, some of the lymphoid hyperplastic spleens, and some histologically normal spleens. (pnas.org)
  • Reverse transcriptase-polymerase chain reaction (RT-PCR) performed for both samples gave a product of 443-bp amplifying the highly conserved "N"-region gene of virus confirming the rabies infection in both cases. (researcherslinks.com)
  • IMSEAR at SEARO: Primary endobronchial synovial sarcoma confirmed by SYT-SSX1 fusion gene transcript by reverse transcriptase polymerase chain reaction. (who.int)
  • The reverse transcriptase polymerase chain reaction (RT-PCR) is a sensitive technique that can detect prostate-specific messenger RNA in circulating blood. (openrepository.com)
  • A study was conducted to apply reverse transcriptase-polymerase chain reaction (RT-PCR) technique for the confirmative diagnosis of canine distemper in dogs. (unud.ac.id)
  • Comparative Evaluation of Immunochromatographic and Reverse Transcriptase Polymerase Chain Reaction based tests for Diagnosis of Canine Distemper. (thefreelibrary.com)
  • Various methods have been suggested for the diagnosis of COVID-19, including chest computed tomography (CT) scans and reverse transcriptase-polymerase chain reaction (RT-PCR) assays. (paho.org)
  • False-Negative Results of Real-Time Reverse-Transcriptase Polymerase Chain Reaction for Severe Acute Respiratory Syndrome Coronavirus 2: Role of Deep-Learning-Based CT Diagnosis and Insights from Two Cases. (cdc.gov)
  • The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. (unifesp.br)
  • Real-time quantitative polymerase-chain-reaction (qRT-PCR) is able to detect HER2 overexpression. (biomedcentral.com)
  • To detect MDR1 mRNA, reverse transcriptase-polymerase chain reaction (RT-PCR) was performed in all samples. (elsevier.com)
  • Real-time RT-PCR has the ability to measure several fluorophores in one well and permits multiplex assays, so it can be used to detect different target sequences simultaneously in one reaction [ 4 ]. (biomedcentral.com)
  • Sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction detecting feline coronavirus mutations in effusion and serum/plasma of cats to diagnose feline infectious peritonitis. (physiciansweekly.com)
  • Eligibility criteria and data analysis Eligible studies measured sensitivity or specificity, or both of a covid-19 serological test compared with a reference standard of viral culture or reverse transcriptase polymerase chain reaction. (bmj.com)
  • Reverse-transcriptase polymerase chain reaction (RT-PCR) is a widely used method that enables transcriptional profiling and sequencing analysis on bulk populations of cells. (eurekamag.com)
  • Diagnostic performance of CT and reverse transcriptase-polymerase chain reaction for coronavirus disease 2019: a meta-analysis. (paho.org)
  • Reverse transcriptase-polymerase chain reaction analysis on 10 individual ruptured and 10 individual stable plaques showed a striking consistency of expression for the clones SSH6, present in 8 ruptured and 2 stable plaques, and perilipin, expressed in 8 ruptured plaques and completely absent in stable plaques. (ahajournals.org)
  • To validate the reproducibility of expression of these clones, reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis was performed on a larger series of ruptured (n=10) and stable (n=10) plaques. (ahajournals.org)
  • Reverse Transcriptase Polymerase Chain Reaction" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (sickkids.ca)
  • Is the Subject Area "Reverse transcriptase-polymerase chain reaction" applicable to this article? (plos.org)
  • Our findings suggest that such peptides may be used in reversing or halting the neurodegenerative process observed in neurodegenerative diseases, such as the peripheral neuropathy associated with type 2 diabetes mellitus and Alzheimer's and Parkinson's diseases. (aspetjournals.org)
  • This graph shows the total number of publications written about "Reverse Transcriptase Polymerase Chain Reaction" by people in this website by year, and whether "Reverse Transcriptase Polymerase Chain Reaction" was a major or minor topic of these publications. (sickkids.ca)
  • Whilst several groups have demonstrated Paramyxoviruses using techniques such as in situ hybridisation (ISH), reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ-RT-PCR (IS-RT-PCR), others have found no evidence of viruses using only RT-PCR. (nih.gov)
  • Purpose: We evaluate the diagnostic use of cytokeratin 20 messenger (m) RNA quantitation in urine as a marker of urothelial transitional cell carcinoma using the real time reverse transcriptase polymerase chain reaction (RT-PCR). (fujita-hu.ac.jp)
  • The nucleic acid test or genetic sequencing serves as the gold standard method for confirmation of infection, yet several recent studies have reported false-negative results of real-time reverse-transcriptase polymerase chain reaction (rRT-PCR). (cdc.gov)
  • 1997. Competitive reverse-transcriptase-polymerase chain reaction shows that dietary zinc supplementation in humans increases mononuclear cell metallothionein mRNA levels. (koreascience.or.kr)
  • To investigate 15 respiratory viruses in children with acute respiratory tract infections [ARTIs] using multiplex reverse- transcriptase polymerase chain reaction [RT-PCR], and to analyze the clinical and epidemiological features of these viruses . (bvsalud.org)
  • Enrolled children had nasal/throat swabs tested for influenza by reverse transcriptase-polymerase chain reaction and their medical records reviewed. (aappublications.org)
  • In this report, we describe a novel droplet-based microfluidic system for performing ~50000 single-cell RT-PCR reactions in a single experiment while consuming a minimal amount of reagent. (eurekamag.com)
  • PCR carries out one reaction per single sample. (wikipedia.org)
  • dPCR also carries out a single reaction within a sample, however the sample is separated into a large number of partitions and the reaction is carried out in each partition individually. (wikipedia.org)
  • Below are the most recent publications written about "Reverse Transcriptase Polymerase Chain Reaction" by people in Profiles. (sickkids.ca)
  • The purpose of this study is to demonstrate the efficacy of active study vaccine in the prevention of reverse transcriptase polymerase chain reaction (RT-PCR) confirmed respiratory syncytial virus (RSV)-mediated lower respiratory tract disease (LRTD), when compared to placebo. (centerwatch.com)
  • In this study, three lineages of LMP1 transgenic mice were established with LMP1 expressed under the control of the Ig heavy chain promoter and enhancer. (pnas.org)
  • For example, if Sample A, when assayed in 1 million partitions, gives one positive reaction, it does not mean that the Sample A has one starting molecule. (wikipedia.org)
  • The PCR solution is divided into smaller reactions and are then made to run PCR individually. (wikipedia.org)