Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Escherichia coli O157:H7 has been recognized since 1982 as a serious human pathogen spread by contaminated food and water. Pulsed-field gel electrophoresis has proven useful for identification of specific isolates/strains of this organism. Hemolytic uremic syndrome (HUS), generally occurring in children or the aged, is the most severe sequela associated with E. coli O157:H7 infection. The presently described work was designed to compare the genomic profile of isolates known to have caused HUS with those having had no such involvement. We asked the question: "Can we develop the means to recognize an 'HUS-prone' E. coli isolate and thereby alert medical personnel to the increased risk?" Twenty-two HUS-related and 10 HUS-unrelated E. coli O157:H7 samples were chosen for genomic analysis. Isolates were cultured overnight prior to being embedded in agarose gel plugs. Plugs were digested, using Xbal restriction endonuclease, and subjected to pulsed-field gel electrophoresis (PFGE) for 20 hours. Gels were stained with ethidium bromide, photographed under ultraviolet light, and Southern blotted. Radiolabeled toxin gene probes were used for hybridization assays. The two classes of isolates were compared by optical imaging software. A computer-generated dendrogram, based on restriction profiles, offered strong initial evidence that the HUS sequela may be produced by a particularly virulent and identifiable clone. The predictive value of this finding appears to be substantial.
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PMID:Evaluation of DNA "fingerprinting" for predicting the potential of E. coli O157:H7 isolates to cause hemolytic uremic syndrome (HUS). 919 12

To construct the recombinant adenovirus vector expressing specific siRNA for rat retinoic acid receptor-beta (RARbeta) gene, and to detect its effect on RARbeta expression and neuronal differentiation of all-trans retinoic acid (ATRA) treated mesenchymal stem cells (MSCs). First, we designed four pairs of siRNA sequence for rat RARbeta gene and annealed complementary oligonucleotides in vitro, then cloned double-stranded DNA in pSES-HUS vector containing U6/H1 double-promoter and recombinated with the backbone vector to construct pAd-siRARbeta plasmid. We infected MSCs by using adenovirus Ad-siRARbeta which was packaged in HEK293 cell line, then performed Real-time, Western blotting and immunoflourencence to detect the expression of RARbeta. We used combination of ATRA and MNM to induce MSCs into neural-like cells, then performed Real-time PCR and immunoflourencence to detect neuronal specific markers of induced neural cells. By using PCR, endonuclease cutting and gene sequencing, we confirmed that the target genes were correctly cloned in adenovirus vector. We could observe more than 60% RFP-positive MSCs at 24 h after adenovirus infection. The expression of RARbeta was significantly increased to 16.5 +/- 2.34 fold in ATRA treated MSCs (P < 0.05) and located in nucleus. Three of four pairs siRNA could effectively inhibit the expression of RARbeta with inhibition efficacy of (66.26 +/- 9.12)%, (48.70 +/- 5.78)%, (64.09 +/- 0.53)% (P < 0.05), especially siRNA-pool group with inhibition efficacy of (78.09 +/- 4.24)% (P < 0.01). Combination of ATRA and MNM induced MSCs into neural-like cells which expressed neuronal specific markers, Nestin, NSE, MAP-2, and Tau. Immunoflourencence result showed that about 50-88 present of cells were positive for Nestin, NSE, Tjul, however, adenovirus medicated expression of siRARbeta could effectively inhibit the expression level of neural specific proteins and the ratio of positive stained cells (P < 0.05). Therefore, we successfully constructed the recombinant adenovirus vector containing siRNA for rat RARP gene, adenovirus could effectively infect MSCs and inhibit the expression of induced RARbeta in ATRA treated MSCs, then inhibit neuronal differentiation of MSCs.
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PMID:[Recombinant adenovirus expressing siRNA is generated to inhibit the expression of RARbeta in rat mesenchymal stem cells treated by all-trans retinoic acid]. 2291 1