Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Sentinel lymph node (SLN) status is highly predictive of overall axillary lymph node involvement in breast cancer. Historically, SLN-positive patients have undergone axillary lymph node dissection in a second surgery. Intraoperative SLN analysis could reduce the cost and complications of a second surgery; however, existing histopathological methods lack standardization and exhibit poor sensitivity. Rapid molecular methods may lead to improved intraoperative diagnosis of SLN metastasis. In this study, we used a genome-wide gene expression analysis of breast and other tissues to identify seven putative markers for detecting breast cancer metastasis. We assessed the utility of these markers for identifying clinically actionable metastases in lymph nodes through reverse transcriptase-polymerase chain reaction analysis of SLNs from 254 breast cancer patients. Polymerase chain reaction signals were compared to pathology on a per-patient basis. The optimal two-gene combination, mammaglobin and cytokeratin 19, detected clinically actionable metastasis in breast SLNs with 90% sensitivity and 94% specificity. Application of stringent criteria for identifying presumptive hematoxylin- and eosin-positive samples increased sensitivity and specificity to 91 and 97%, respectively. This study represents the first comprehensive demonstration of the utility of gene expression markers for detecting clinically actionable breast metastases. An intraoperative molecular assay using these markers has the potential to significantly reduce second surgeries for patients undergoing SLN dissection.
...
PMID:Identification and characterization of optimal gene expression markers for detection of breast cancer metastasis. 1604 4

We previously reported on a de novo designed protein "milk bundle-1Trp" (MB-1Trp) as a source of selected essential amino acids (EAA) for ruminant feeding. Here, we attempt to express this de novo designed protein in alfalfa. The microbial version of the gene encoding the protein was modified in order to achieve two expression strategies in transgenic alfalfa plants. Chimeric MB-1Trp genes alone or fused to a signal peptide and an endoplasmic reticulum retention sequence were introduced into alfalfa via Agrobacterium-mediated transformation. Polymerase chain reaction and reverse transcriptase polymerase chain reaction analysis performed on individual transgenic lines demonstrated that the MB-1Trp gene was correctly integrated and transcribed into mRNA. However, under our conditions, it was impossible to detect MB-1Trp protein expression in any of the transgenic plants analyzed. In order to assess MB-1Trp stability in alfalfa, Escherichia coli-derived MB-1Trp was incubated with proteins extracted from leaves of a non-transgenic plant. This study revealed a high susceptibility of mature MB-1Trp to alfalfa proteases, which may have contributed to its lack of accumulation.
...
PMID:The de novo designed nutritive protein MB-1Trp does not resist proteolytic degradation in alfalfa leaves. 1638 25

Cisplatin is a widely used chemotherapeutic agent whose dose-limiting side effects include ototoxicity and nephrotoxicity. Recent evidence indicates that cisplatin induces the expression of a novel protein, kidney injury molecule-1, in the renal proximal tubular epithelium to aid in regeneration. In this study, we determined whether kidney injury molecule-1 is expressed in the cochlea and is induced by cisplatin. Using reverse transcriptase polymerase chain reaction techniques, we have now identified kidney injury molecule-1 in the rat cochlea and in three different mouse transformed hair cell lines. Administration of cisplatin to rats produced hearing loss and induced kidney injury molecule-1 mRNA in the rat cochlea. Pretreatment of rats with lipoic acid, a scavenger of reactive oxygen species, significantly reduced cisplatin-induced hearing loss and kidney injury molecule-1 expression. Cisplatin also increased the expression of cochlear NOX3 mRNA, a member of the superoxide generating NADPH oxidase family of proteins recently identified in the cochlea, inhibition of which decreased kidney injury molecule-1 expression. Polymerase chain reaction performed on different regions of the cochlea indicated the presence of kidney injury molecule-1 mRNA in the lateral wall, organ of Corti and spiral ganglion. This distribution was confirmed by immunocytochemistry. Taken together, these data identify kidney injury molecule-1 as a novel cochlear injury molecule, whose expression is regulated by reactive oxygen species generated via the NADPH oxidase pathway.
...
PMID:Expression of the kidney injury molecule 1 in the rat cochlea and induction by cisplatin. 1646 36

Nucleic acid aptamers to HIV-1 reverse transcriptase (RT) are potent inhibitors of DNA polymerase function in vitro, and they have been shown to inhibit viral replication when expressed in cultured T-lymphoid lines. We monitored RT inhibition by five RNA pseudoknot RNA aptamers in a series of biochemical assays designed to mimic discrete steps of viral reverse transcription. Our results demonstrate potent aptamer inhibition (IC50 values in the low nanomolar range) of all RT functions assayed, including RNA- and DNA-primed DNA polymerization, strand displacement synthesis, and polymerase-independent RNase H activity. Additionally, we observe differences in the time dependence of aptamer inhibition. Polymerase-independent RNase H activity is the most resistant to long term aptamer suppression, and RNA-dependent DNA polymerization is the most susceptible. Finally, when DNA polymerization was monitored in the presence of an RNA aptamer in combination with each of four different small molecule inhibitors, significant synergy was observed between the aptamer and the two nucleoside analog RT inhibitors (azidothymidine triphosphate or ddCTP), whereas two non-nucleoside analog RT inhibitors showed either weak synergy (efavirenz) or antagonism (nevirapine). Together, these results support a model wherein aptamers suppress viral replication by cumulative inhibition of RT at every stage of genome replication.
...
PMID:Differential susceptibility of HIV-1 reverse transcriptase to inhibition by RNA aptamers in enzymatic reactions monitoring specific steps during genome replication. 1679 47

During the period from January 2000 to August 2005 a total of 204 outbreaks of norovirus gastroenteritis were diagnosed at the Norwegian Institute of Public Health. A clear increase in the norovirus activity was seen in healthcare institutions during the winter seasons. Polymerase sequence analysis of norovirus strains from 122 outbreaks showed that 112 were caused by GII strains (91.8%). Two norovirus variants seen during the study period-GIIb and GII.4-were predominant between January 2000 and September 2002, whereas GII.4 was predominant from September 2002 onward. The highest norovirus activity was seen during the 2002-2003 and 2004-2005 seasons with the emergence of new GII.4 variants. This study describes the molecular epidemiology of norovirus strains circulating in Norway during the five previous seasons and compares four norovirus real-time reverse transcriptase PCR assays. A suitable assay for routine diagnostics is suggested.
...
PMID:Molecular epidemiology of norovirus outbreaks in Norway during 2000 to 2005 and comparison of four norovirus real-time reverse transcriptase PCR assays. 1702 Oct 99

Bluetongue virus (BTV) is the prototype of the member of the Orbivirus genus within the family Reoviridae. The BTV serogroup contains 24 serotypes. Traditionally, viruses have been isolated in cultured cells, suckling mouse brain or embryonated chicken eggs before their identification and biochemical, antigenic and biological characterization. These procedures are time-consuming and may fail to detect low levels of infectious virus or strains of BTV which fail to replicate in eggs, mice or tissue culture. In the past decade, traditional procedures for virus characterization, such as ELISA and serum neutralisation with serotype-specific antisera, have been supplemented by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing. A number of procedures have been developed to detect the presence of BTV antigens or nucleic acids. RT-PCR technique has appeared to be a powerful tool in the field of BTV diagnosis. Polymerase chain reaction techniques may be used not only to detect the presence of viral nucleic acid but also to 'serogroup' orbiviruses and provide information on the serotype and possible geographical source (topotype or genotype) of BTV isolates within a few days of receipt of a clinical sample such as infected sheep blood. Real-time PCRs have recently been developed.
...
PMID:Bluetongue: characterization of virus types by reverse transcription-polymerase chain reaction. 1705 94

A subtracted cDNA library was used to identify specific genes that increase in post-mortem muscle of rainbow trout (Oncorhynchus mykiss) during on-ice storage. Of the 200 cDNAs analyzed, 82 had significant homologies to previously identified genes from salmonids and other species such as homologues of troponin I, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and so on, whereas 40 had no significant homologies and were designated as unknown. Comparison of gene expression profiles by dot blot hybridization confirmed an increase or induction of mRNA in the muscle after 3 h of on-ice storage compared to that at 0 h after death. Real-time reverse transcriptase-Polymerase Chain Reaction analysis showed that troponin I and GAPDH mRNAs were increased by 24 h and, in particular, that the change in troponin I mRNA was greater than that of GAPDH mRNA. These results suggest that the increased mRNAs in rainbow trout muscle occurred by transcription immediately after death.
...
PMID:Post-mortem changes in gene expression of the muscle tissue of rainbow trout, Oncorhynchus mykiss. 1714 27

In this study, it was shown that the TsVP gene [vacuolar H+-pyrophosphatase (V-H+-PPase) gene from a dicotyledonous halophyte Thellungiella halophila] could be transferred into the monocotyledonous crop maize (Zea mays L.), and that the heterologous expression of the transgene improved the drought resistance of transgenic plants. Polymerase chain reaction amplification and Southern blotting confirmed the existence of the foreign gene in transformed plants and their progeny. Expression differences of the TsVP gene in different transgenic lines were monitored by reverse transcriptase-polymerase chain reaction. The measurement of isolated vacuolar membrane vesicles from the TsVP transgenic and wild-type (WT) plants demonstrated that the transgenic plants had higher V-H+-PPase activity, and the performance of maize-expressed TsVP in response to osmotic/drought stress was better in lines with higher V-H+-PPase activity. Transgenic plants showed a higher percentage of seed germination, better developed root systems and greater biomass, greater solute accumulation and less cell membrane damage relative to WT plants under osmotic stress. After drought stress treatment, transgenic plants showed less growth retardation and shorter anthesis-silking interval, and produced much larger grain yields, than WT plants. It was concluded that the high V-H+-PPase activity of transgenic maize improved the drought resistance of plants. This report provides a feasible approach to increase monocotyledonous crop yields under conditions of soil water deficit by the heterologous expression of TsVP.
...
PMID:Heterologous expression of the TsVP gene improves the drought resistance of maize. 1799 58

Full-length starch branching enzyme II (SBE, EC 2.4.1.18) cDNA from mungbean ( Vigna radiata L. cv. Tainan no. 5), VrsbeII, was cloned, characterized, and expressed as an active enzyme in Escherichia coli . Gene-specific primers first amplified an internal cDNA by reverse transcriptase Polymerase Chain Reaction (RT-PCR), followed by obtaining 5' and 3' fragments by RT-PCR and rapid amplification of cDNA ends (RACE). VrsbeII possesses a complete open reading frame (ORF) of 2571 bp, and the deduced polypeptide includes the common catalytic (beta/alpha)(8)-barrel domain and conserved regions of the alpha-amylase family. Phylogenetic analysis classified VrsbeII into SBE family A. Its partial 3D structure and functional features were predicted. VrsbeII has a shorter N-terminal among SBEs; however, two 6 bp (CCAGTT) direct repeat sequences (DRS) were found. A 24 bp shortened VrsbeII at the 3' end, skipping one DRS, was ligated into pET21b vector and expressed as His(6)-rVrSBEII in E. coli BL21 (DE3) cells. The optimal expression condition for rVrSBEII was evaluated and detected by Western blot with a molecular size of 108 kDa and activity of 6.4 U/mg.
...
PMID:Cloning, characterization, and expression of mungbean ( Vigna radiata L.) starch branching enzyme II cDNA in Escherichia coli. 1914 23

Polymerase chain reaction (PCR)-based approaches to the detection, differentiation and characterization of avian pathogens continue to be developed and refined. The PCRs, or reverse transcriptase-PCRs, may be general, designed to detect all or most variants of a pathogen, or to be serotype, genotype or pathotype specific. Progress is being made with respect to making nucleic acid approaches more suitable for use in diagnostic laboratories. Robotic workstations are now available for extraction of nucleic acid from many samples in a short time, for routine diagnosis. Following general PCR, the DNA products are commonly analyzed by restriction endonuclease mapping (restriction fragment length polymorphism), using a small number of restriction endonucleases, based on a large body of sequence data. Increasingly, however, nucleotide sequencing is being used to analyze the DNA product, in part due to the expanding use of non-radioactive sequencing methods that are safe and enable high throughout. In this review, I highlight some recent developments with many avian viruses: Newcastle disease virus; circoviruses in canary and pigeon; infectious bursal disease virus (Gumboro disease virus); avian adenoviruses, including Angara disease/infectious hydropericardium virus, haemorrhagic enteritis virus of turkeys, and egg drop syndrome virus; avian herpesviruses, including infectious laryngotracheitis virus, duck plague virus, psittacine herpesvirus (Pacheco's parrot disease virus), Marek's disease virus and herpesvirus of turkeys; avian leukosis virus (associated with lymphoid leukosis or myeloid leukosis, and egg transmission); avian pneumoviruses (turkey rhinotracheitis virus); avian coronaviruses, including infectious bronchitis virus, turkey coronavirus and pheasant coronavirus; astrovirus, in the context of poult enteritis and mortality syndrome, and avian nephritis virus; and avian encephalomyelitis virus, a picornavirus related to hepatitis A virus.
...
PMID:Innovation and discovery: the application of nucleic acid-based technology to avian virus detection and characterization. 1918 52


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---