Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Focal epilepsies in young patients are frequently associated with differentiated glioneuronal tumours. Dysplastic neurones represent a characteristic neuropathological feature of gangliogliomas, the most common entity encountered in this group. Here, we have analysed two major components of the reelin pathway involved in neuronal migration and cortical development, that is, p35 and disabled-1 (dab1), in gangliogliomas. Genomic structures of human dab1 and p35 were identified 'in silico' using the HTGS databank, NCBI BLAST 2.1. DNA sequence analysis was carried out in gangliogliomas obtained from 29 epilepsy patients vs. peripheral blood DNA from non-affected control individuals (n = 100). Gene expression of dab1 and p35 was determined by real-time RT-PCR (reverse transcriptase polymerase chain reaction) in gangliogliomas (n = 14) vs. non-neoplastic central nervous system tissue (n = 20). The human dab1 gene contains 13 coding exons and is located on chromosome 1p31-32. A single coding exon constitutes the human p35 gene, which is located on chromosome 17q11.2. A novel homologueous genomic region on chromosome 2 has to be taken into account for future studies on p35. One ganglioglioma patient showed a unique polymorphism in the p35 gene. The single base exchange (C to A) at nucleotide 904 of the p35 cDNA (GenBank X80343, start ATG, codon 302) results in a leucine-isoleucine amino acid substitution. No mutations of the dab1 and p35 genes in gangliogliomas were observed. However, significantly lower levels of dab1 and p35 gene transcripts were detected in gangliogliomas compared to controls (dab1 28.24%, t-test P < 0.001; p35 21.28%, t-test P < 0.001, in gangliogliomas vs. controls). Our data suggest that mutational events of dab1 and p35 are not involved in the molecular pathogenesis of gangliogliomas. A potential functional role of these developmentally regulated genes for the formation of epileptogenic glioneuronal lesions remains to be elucidated.
 ...
PMID:The reelin pathway components disabled-1 and p35 in gangliogliomas--a mutation and expression analysis. 1517 76

The immune system of Indian water buffalo (Bubalus bubalis), one of the major dairy animals of the tropics, has received little attention. cDNAs encoding the two subunits of the heterodimeric interleukin (IL)-12 of Indian water buffalo were isolated from concanavalin A-stimulated lymphocytes. The 710-bp p35 and 1012-bp p40 subunits were amplified by reverse transcriptase polymerase chain reaction (RT-PCR), cloned, sequenced and compared with other ruminant sequences. The IL-12 p35 subunit cDNA had nine nucleotide variations and shared 98.1% amino acid identity with the cattle IL-12 p35. The IL-12 p40 cDNA had 13 nucleotide variations and had 97.5% amino acid identity with the cattle IL-12 p40. Both the subunits showed strict conservation in the predicted secondary structure and critical amino acid residues compared with other ruminant IL-12 molecules. Buffalo IL-12 p40 recombinant protein expressed in Escherichia coli cross-reacted with cattle anti-IL-12 p40 monoclonal antibody. Our study indicates a high level of conservation of this key cytokine among ruminants.
 ...
PMID:Interleukin-12 subunits p35 and p40 of Indian water buffalo (Bubalus bubalis) maintain high sequence homology with those of other ruminants. 1593 23

Inflammatory bowel disease (IBD) is a common condition in cats characterised by infiltration of inflammatory cells into the intestinal mucosa. In this study, real-time reverse transcriptase polymerase chain reaction (RT-PCR) was used to quantify cytokine messenger RNA (mRNA) expression in intestinal biopsies from cats. Biopsies were collected from seven cats with chronic diarrhoea and histologically confirmed IBD, five cats with chronic diarrhoea due to non-IBD gastrointestinal (GI) disease, and nine clinically normal cats with or without subclinical inflammatory changes in small intestine. Real-time RT-PCR was developed for quantification of mRNA encoding interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p35 and p40), IL-18, tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a 'housekeeper' gene. All real-time PCR efficiencies were>90% (range 90.4-102%) with correlation coefficients >0.99 (range 0.998-1). The results of the study were analyzed on the basis of either clinical presentation or histopathological evidence of intestinal inflammation. The former analysis showed that mRNA encoding IL-10 and TGF-beta (immunoregulatory cytokines), and IL-6, IL-18, TNF-alpha and IL-12 p40 (Th1 and pro-inflammatory cytokines) was significantly higher in clinically normal cats and cats with IBD when compared to cats with other GI diseases. IL-5 mRNA was significantly higher in cats with IBD compared to clinically normal cats. IL-2 mRNA was significantly lower in cats with non-IBD GI disease than in clinically normal cats. Analysis on the basis of histopathological change revealed that cats with intestinal inflammation had significantly more transcription of genes encoding IL-6, IL-10, IL-12p40, TNF-alpha and TGF-beta than those with normal intestinal morphology. The results suggest that immune dysregulation plays a role in feline IBD and that IBD in cats has a complicated pathogenesis with both pro-inflammatory and immunoregulatory features.
 ...
PMID:Measurement of cytokine mRNA expression in intestinal biopsies of cats with inflammatory enteropathy using quantitative real-time RT-PCR. 1687 76

In order to investigate the mRNA expression and function of interleukin-23 (p19/p40) and interleukin-12 (p35/p40) in the psoriatic lesion, no-lesion and normal human skin, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-23 (p19/p40) and IL-12 (p35/p40). The results showed that the expression of IL-23p19 mRNA and p40 (IL-12/IL-23) mRNA were higher in psoriatic lesion than those of non-lesional skin and normal skin. The levels of IL-23p19 mRNA and p40 (IL-12/IL-23) mRNA were higher in psoriatic non-lesional skin than normal skin. However, no significant difference was found in the level of IL-12p35 mRNA among the psoriatic lesional skin, non-lesional skin and normal skin. It was suggested that IL-23 might be more important in the pathogenesis of psoriasis than IL-12.
 ...
PMID:The expression of interleukin-23 (p19/p40) and interleukin-12 (p35/p40) in psoriasis skin. 1735 9

Feline allergic skin disease is thought to be associated with dermal infiltration of Th(2) lymphocytes and synthesis of associated cytokines. In this study, real-time RT-PCR assays were developed to measure feline interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL12 (p35 and p40), IL-18, tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), interferon-gamma (IFN-gamma) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in the skin of healthy control cats, and in the lesional and non-lesional skin of cats with allergic skin disease. Total RNA was extracted from skin biopsies using the RNeasy Mini Kit with on-column and in-solution DNase digestion steps. cDNA was synthesised using Improm-II reverse transcriptase and random hexamers. Real-time PCR was carried out using an iCycler IQ system (Bio-Rad), and gene-specific primers were designed to span an exon/exon junction of each cytokine gene. Taq-man probes were used to add specificity to the system. Messenger RNA from the housekeeping gene GAPDH was used for normalisation of the cytokine threshold cycle. The eleven cytokine mRNA transcripts quantified were present at varying levels, but there was no apparent difference in expression between normal, non-lesional and lesional skin. TGF-beta represented the most abundant transcript while IL-4, IL-5, IL-6, IL-10, IL-12, IL-18 and TNF-alpha were present at levels approximately 1000-fold less. IL-2 and INF-gamma represented the least abundant templates with no detectable copies in most RNA samples. This quantitative analysis of cytokine mRNA expression in feline skin biopsies has suggested that there is not a simple Th(2) bias in lesional skin of cats with allergic dermatopathies.
 ...
PMID:Quantitative real-time RT-PCR measurement of cytokine mRNA expression in the skin of normal cats and cats with allergic skin disease. 1819 Dec 30

The heterodimeric cytokine IL-12 (composed of a p35 and a p40 subunit) is produced primarily by monocytes, macrophages and B cells. In vitro and in vivo experiments have demonstrated the crucial role of IL-12 in initiating and establishing both innate immunity and T cell-mediated resistance to intracellular pathogens, including Leishmania donovani, Toxoplasma gondii, Listeria monocytogenes, and Mycobacterium tuberculosis. Assessment of cytokine expression has thus become crucial to understand host responses to infections. In this study, by using the reverse transcriptase-real time PCR we developed a highly specific and sensitive assay to quantitatively evaluate IL-12p40 mRNA transcription levels in peripheral blood mononuclear cells (PBMCs) stimulated with PHA vs. unstimulated cells. We also used the ELISA to evaluate bioactive IL-12 release in culture supernatants. We provide evidence that IL-12 p40 mRNA levels were significantly up-regulated in PHA-activated PBMCs. These results were correlated with data of IL-12 levels obtained by ELISA.
 ...
PMID:Quantitative evaluation of interleukin-12 p40 gene expression in peripheral blood mononuclear cells. 1830 39

Narrow-leafed lupin (NLL; Lupinus angustifolius) is a recently domesticated but anciently propagated crop with significant value in rotation with cereals in Mediterranean climates. However, several fungal pathogens, traditionally termed necrotrophs, severely affect broad-acre production and there is limited genetic resistance in the NLL germplasm pool. Symptoms of many of these diseases appear as localized areas of dead cells exhibiting markers of programmed cell death. Based on our previous research, we hypothesized that engineered expression of the baculovirus anti-apoptotic p35 gene might reduce symptoms of these diseases. Using Agrobacterium tumefaciens-mediated transformation of a cultivar highly susceptible to several pathogens, 14 independent NLL lines containing both the p35 and bar genes were obtained (p35-NLL). Integration and expression of the transgenes were confirmed by polymerase chain reaction (PCR), progeny testing, Southern blot, Northern blot and reverse transcriptase-PCR analyses. Fecundity and nodulation were not altered in these lines. Third or fourth generation p35-NLL lines were challenged with necrotrophic fungal pathogens (anthracnose in stem and leaf, and Pleiochaeta root rot and leaf brown spot) in controlled environment conditions. Several p35-NLL lines had significantly reduced disease symptoms. Interestingly, as with natural resistance, no single line was improved for all three diseases which possibly reflecting spatial variation of p35 expression in planta. These data support an alternative molecular definition for 'necrotrophic disease' in plants and suggest new routes for achieving resistance against a range of pathogens.
 ...
PMID:Significant reduction of fungal disease symptoms in transgenic lupin (Lupinus angustifolius) expressing the anti-apoptotic baculovirus gene p35. 1975 37


<< Previous 1 2 3