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:3.1.27.5 (RNase)
17,967 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Basic fibroblast growth factor (bFGF) stimulates bone formation in vitro and in vivo. The purpose of this study was to determine changes in gene expression for bone matrix proteins, growth factors, and cytokines associated with the stimulatory effects of bFGF on bone formation in aged ovariectomized (ovx) rats. At 3 months of age, female Sprague-Dawley rats were sham-operated (sham) or ovariectomized (ovx), then maintained untreated for 1 year. At 15 months of age, baseline (BSL) sham and ovx rats were killed. All other rats received daily intravenous injections of bFGF (200 microg/kg) or vehicle (veh) for 14 days. Lumbar vertebrae were processed for quantitative bone histomorphometry or molecular analyses. Ovariectomy decreased vertebral cancellous bone volume by approximately 33% and increased most indices of bone turnover. Treatment of aged ovx rats with bFGF for only 14 days significantly increased cancellous bone volume compared with vehicle treatment of ovx rats, but this variable remained lower than in sham + veh rats. Osteoid volume, osteoblast surface, and osteoid surface were markedly increased, and osteoclast surface was significantly decreased in ovx + bFGF rats compared with sham + veh and ovx + veh rats. Northern analyses revealed that mRNA levels for osteocalcin and type I collagen, relative to 18S RNA, were significantly higher in ovx + bFGF rats than in ovx + veh rats by a factor of >10. RNase protection assays revealed that insulin-like growth factor (IGF-I) mRNA levels, relative to L32 housekeeping gene, were also significantly higher, by nearly a factor of 3, in ovx + bFGF rats than in ovx + veh rats. Treatment of ovx rats with bFGF did not appear to affect message levels for transforming growth factor-beta (TGF-beta), interleukin-6 (IL-6), and interferon-gamma (IFN-gamma). These in vivo results suggest that bFGF treatment upregulates gene expression for IGF-I, which may mediate, at least in part, the increased gene expression for bone matrix proteins and the bone anabolic effects of bFGF in aged ovx rats.
...
PMID:Changes in gene expression associated with the bone anabolic effects of basic fibroblast growth factor in aged ovariectomized rats. 1211 Apr 27

A broad-host range metabolic cassette has been designed that, under the control of the Ptac promoter, expresses the sytABCD catabolic genes from Pseudomonas sp. Y2, which are responsible for the transformation of styrene into phenylacetic acid (styrene upper pathway). This novel cassette confers to phenylacetic acid-degrading bacteria the ability to grow efficiently on styrene as the sole carbon and energy source. By combining both the sty cassette and the archetypal pWW0 TOL plasmid into the well-known Pseudomonas putida F1 aromatic biodegrader, we have constructed a novel derivative strain that shows one of the largest degradative potentials so far described for aromatic hydrocarbons, because it is able to use BTEX compounds (benzene, toluene, ethylbenzene and xylenes) and styrene as a source of carbon and energy. Furthermore, the sty cassette was engineered within a mini-transposon and endowed with a gene containment system, based on the toxic effect of the colicin E3 RNase, to reduce its lateral spread to other hosts. This contained cassette lacks defined transcriptional regulatory signals and, thus, it becomes an alternative strategy to select recombinant strains that efficiently express the desired phenotype from housekeeping regulatory elements.
...
PMID:Design of catabolic cassettes for styrene biodegradation. 1290 58

The human translation termination factor 1 (ETF1) gene encodes a class-1 release factor, eRF1, which catalyses termination of protein synthesis at all three stop codons. In this report, we describe the functional organization of the 5'-region of the gene. Primer extension and ribonuclease protection mapping revealed three transcription start sites clustered within approximately 10 bp. DNase I-hypersensitive site analysis identified five hypersensitive sites, one of which was located downstream of the initiation start sites. We used transient expression assays to define the 5'-regulating regions and in vivo and in vitro footprinting analysis to identify potential cis-acting regulatory elements. A basal promoter, spanning nucleotides -210/+117, contained no TATA box but a putative initiator element (Inr) and multiple potential Sp1/Sp3 binding sites, and thus displayed some of the features of a housekeeping gene. An additional upstream promoter containing positive and negative regulatory elements also regulated ETF1 gene expression. Real-time quantitative RT-PCR analysis showed tissue-specific expression of ETF1 transcripts in mouse tissues. Our results are suggestive of a constitutive expression of the human ETF1 gene but with possible cell- and tissue-specific regulation.
...
PMID:Promoter analysis of the human translation termination factor 1 gene. 1456 55

Strenuous resistive breathing induces plasma cytokines that do not originate from circulating monocytes. We hypothesized that cytokine production is induced inside the diaphragm in response to resistive loading. Anesthetized, tracheostomized, spontaneously breathing Sprague-Dawley rats were subjected to 1, 3, or 6 hours of inspiratory resistive loading, corresponding to 45-50% of the maximum inspiratory pressure. Unloaded sham-operated rats breathing spontaneously served as control animals. The diaphragm and the gastrocnemius muscles were excised at the end of the loading period, and messenger ribonucleic acid expression of tumor necrosis factor-alpha, tumor necrosis factor-beta, interleukin (IL)-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IFN-gamma, and two housekeeping genes was analyzed using multiprobe RNase protection assay. IL-6, IL-1beta, and, to lesser extents, tumor necrosis factor-alpha, IL-10, IFN-gamma, and IL-4 were significantly increased in a time-dependent fashion in the diaphragms but not the gastrocnemius of loaded animals or in the diaphragm of control animals. Elevation of protein levels of IL-6 and IL-1beta in the diaphragm of loaded animals was confirmed with immunoblotting. Immunostaining revealed IL-6 protein localization inside diaphragmatic muscle fibers. We conclude that increased ventilatory muscle activity during resistive loading induces differential elevation of proinflammatory and antiinflammatory cytokine gene expression in the ventilatory muscles.
...
PMID:Differential cytokine gene expression in the diaphragm in response to strenuous resistive breathing. 1511 43

Human herpesvirus-8 (HHV-8, also called Kaposi's sarcoma-associated herpesvirus) infectious cycle can be divided into latent and lytic phases. During the latent phase viral gene expression is reduced to a minimum, while during the lytic phase, numerous genes are expressed sequentially. The development of an RNase Protection Assay (RPA) is described that allows the detection and quantitation of three latent (ORFs 71-72-73) and three lytic (ORFs 74-K4-K2) HHV-8 genes as well as two cellular housekeeping gene transcripts (glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and cyclophilin) for normalization purposes is described. The RPA was validated using 293T cells transfected with corresponding HHV-8 expression vectors and using resting and phorbol ester-butyric acid-activated BC-3 and BCBL-1 cells. The results obtained indicate that this RPA is specific, sensitive and allows for the simultaneous monitoring of HHV-8 latent and lytic genes expression. HHV-8-RPA is therefore a useful technique to monitor the status of HHV-8 infection in infected cells (latent versus lytic) by comparing and quantitating multiple viral transcripts expression from a single RNA sample.
...
PMID:Multiplex detection and quantitation of latent and lytic transcripts of human herpesvirus-8 using RNase Protection Assay. 1548 14

The growth-promoting effect of growth hormone (GH) is primarily mediated by insulin-like growth factor-1 (IGF-1). The liver is the main source of circulating IGF-I. The authors have used rodent primary hepatocytes for studies on pharmacological intervention of IGF-I mRNA expression. A 96-well nonradioactive IGF-1 mRNA quantification assay was developed, based on the hybridization of sense and antisense RNA probes, to replicate membranes with crude hepatocyte lysates. The sense hybridization was used as an internal standard. The antagonistic properties of a set of GH-receptor binding compounds were evaluated. Two compounds were found to down-regulate IGF-I mRNA. Effects due to metabolic inhibition or toxicity were excluded using a cell proliferation assay. To investigate potential unspecific transcriptional effects, the mRNA levels of the housekeeping genes, beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were determined. Two other GH-regulated genes, cytochrome P450 2C12 (CYP2C12) and a rat homologue to the human alpha1B-glycoprotein (A1BG), were quantified by RNase protection assays and found to be down-regulated, confirming the antagonistic property of 1 compound. In conclusion, a direct filter hybridization assay of hepatocyte lysates using nonradioactive sense and antisense probes can be used for quantitative mRNA measurements and could constitute a valuable tool in screening for pharmacologically active compounds.
...
PMID:Antisense and sense RNA probe hybridization to immobilized crude cellular lysates: a tool to screen growth hormone antagonists. 1580 22

A rapid analysis method for murine endothelin-A (ETA) and endothelin-B (ETB) receptor gene expression levels was established using real-time quantitative reverse transcriptase-polymerase chain reaction. We designed primer pairs and TaqMan probes specific for the two cDNAs and available for mouse and rat systems. The standard curve method was used to examine relative expression. The gene expression levels of ETA and ETB were estimated as gene expression rates by normalizing to the expression of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase. To examine the reproducibility of this assay system, we calculated the intra-assay and interassay coefficients of variation of the gene expression rate and found that a greater than 1.6-fold increase in relative gene expression is detectable as a significant change. ETA and ETB receptor gene expression was found in all 16 organs of mouse and rat examined, and high levels of expression were observed in the lung, uterus, ovary, intestine, and cerebellum. The gene expression patterns essentially agreed with those determined by RNase protection assay, Northern blot, and conventional endpoint polymerase chain reaction. These results show that this new rapid, sensitive, and semi-automated method is accurate, quantitative, and reproducible. This method is also useful for examining regulation of hormone receptor gene expression under physiological conditions in organs.
...
PMID:Real-time polymerase chain reaction quantification of gene expression levels of murine endothelin-A and endothelin-B receptors: gene expression profiles by the standard curve method. 1583 11

Housekeeping genes are widely used as internal controls in a variety of study types, including real time RT-PCR, microarrays, Northern analysis and RNase protection assays. However, even commonly used housekeeping genes may vary in stability depending on the cell type or disease being studied. Thus, it is necessary to identify additional housekeeping-type genes that show sample-independent stability. Here, we used statistical analysis to examine a large human microarray database, seeking genes that were stably expressed in various tissues, disease states and cell lines. We further selected genes that were expressed at different levels, because reference and target genes should be present in similar copy numbers to achieve reliable quantitative results. Real time RT-PCR amplification of three newly identified reference genes, CGI-119, CTBP1 and GOLGAl, alongside three well-known housekeeping genes, B2M, GAPD, and TUBB, confirmed that the newly identified genes were more stably expressed in individual samples with similar ranges. These results collectively suggest that statistical analysis of microarray data can be used to identify new candidate housekeeping genes showing consistent expression across tissues and diseases. Our analysis identified three novel candidate housekeeping genes (CGI-119, GOLGA1, and CTBP1) that could prove useful for normalization across a variety of RNA-based techniques.
...
PMID:Identification of novel universal housekeeping genes by statistical analysis of microarray data. 1739 73

Helicobacter pylori are bacteria with substantial inter-strain variability and phylogenetic reconstructions of sequence data from the organism have common homoplastic sites. Although frequent recombination events have been proposed to contribute to the variation, the effects of nucleotide substitution rate heterogeneities on the reconstruction of H. pylori genealogies have not been studied. We analyzed the substitution pattern of a housekeeping gene, a homologue of the ribonuclease reductase gene (rnr), to characterize rate heterogeneities between 11 H. pylori isolates. Evidence of limited recombination was demonstrated by the Sawyer's runs test, but the homoplasy test and site-by-site compatibility tests indicated frequent recombination events. Within the 1935 nucleotide gene, 292 sites were polymorphic with an average pair-wise difference of 5.01%. Xia's distances for amino acids at non-synonymous codon substitution sites were smaller at homoplastic sites than at sites that were not homoplastic. Transitions were significantly more common among homoplastic than among non-homoplastic nucleotide substitutions. Simulations of evolution with or without recombination indicated the transition/transversion ratio is expected to be higher in homoplastic sites with no recombination. Despite evidence of recombination, analyses of the rnr genealogy does not show a random tree but rather base substitution behaviors characteristic of both recombination and substitution saturation at some sites. Analyses of sequences in the H. pylori multilocus sequence-typing database provided similar evidence for substitution saturation in multiple housekeeping genes.
...
PMID:Rate heterogeneity in the evolution of Helicobacter pylori and the behavior of homoplastic sites. 1857 92

Various techniques for quantification of PCR are available. Most frequently, the densitometric intensities of ethidium bromide-stained PCR products separated in gels are compared after normalizing to the levels of housekeeping gene products such as beta-actin. More precise, but extremely time consuming, is the technique of competitive PCR. Newer methods, such as tracking amplification in real-time, have high start-up and maintenance costs (e.g., TaqMan, Applied Biosystems; LightCycler, Roche; I-Cycler, Bio-Rad). Here, I describe an alternative, simple technique to quantify PCR products by determining the entire phosphate released during PCR. The method can be performed using common laboratory equipment, and the reagents needed are extremely cheap. The method is validated by measuring the induction of inducible nitric oxide synthase gene expression in cell culture and comparing the results with data obtained by LightCycler experiments and RNase protection assays.
...
PMID:Quantification of PCR products by phosphate measurement. 1894 73


<< Previous 1 2 3 4 5 6 7 Next >>

---