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A high throughput assay for inhibitors of HIV-1 protease. Screening of microbial metabolites 总被引:1,自引:0,他引:1
E Sarubbi M L Nolli F Andronico S Stella G Saddler E Selva A Siccardi M Denaro 《FEBS letters》1991,279(2):265-269
A novel method for discovery of HIV-1 protease inhibitors in complex biological samples has been developed. The assay is based on two specific reagents: a recombinant protein constituted by a portion of the HIV-1 Gag polyprotein comprising the p17-p24 cleavage site, fused to E. coli beta-galactosidase, and a monoclonal antibody which binds the fusion protein in the Gag region. Binding occurs only if the fusion protein has not been cleaved by the HIV-1 protease. The assay has been adapted for the screening of large numbers of samples in standard 96-well microtiter plates. Using this method about 12000 microbial fermentation broths have been tested and several HIV-1 protease inhibitory activities have been detected. One of these has been studied in detail. 相似文献
5.
Nucleotide sequence to a teicoplanin resistance gene from Actinoplanes teichomyceticus. 总被引:1,自引:0,他引:1
A DNA fragment, isolated from A. teichomyceticus and able to confer teicoplanin resistance in a sensitive host, has been sequenced. It reveals the presence of two open reading frames (ORFs) positioned on opposite strands, named ORF1 and ORF2. ORF2 seems to be responsible for the acquisition of the resistance character. 相似文献
6.
Gabriella Caruso Maria G. Denaro Lucrezia Genovese 《Marine and Freshwater Behaviour and Physiology》2013,45(4):215-228
Changes occurring after feeding in the digestive enzyme activities of European eel were investigated to provide some insights into the digestive physiology of this fish. Total and specific proteases, amylase and lipase activities were measured using standard biochemical assays over a 24 h cycle in fed eels, compared to starved ones, under the same rearing conditions. In the gastrointestinal tract of fed eels quantitative changes started 4 h after feeding and continued later on; conversely, in starved eels enzyme activities remained unchanged over time. In fed eels, total and specific protease activities showed an overall increasing trend in the intestine, while in the stomach they progressively decreased to values 22–50% lower than those measured at the pre-feeding time; this behaviour probably reflected the progression of digesta along the intestinal tract. The prolonged secretory response of European eel to food ingestion proved its extended activity in the digestive process. 相似文献
7.
The anabolic action of intermittent parathyroid hormone on cortical bone depends partly on its ability to induce nitric oxide‐mediated vasorelaxation in BALB/c mice
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S Gohin A Carriero C Chenu AA Pitsillides TR Arnett M Marenzana 《Cell biochemistry and function》2016,34(2):52-62
There is strong evidence that vasodilatory nitric oxide (NO) donors have anabolic effects on bone in humans. Parathyroid hormone (PTH), the only osteoanabolic drug currently approved, is also a vasodilator. We investigated whether the NO synthase inhibitor L‐NAME might alter the effect of PTH on bone by blocking its vasodilatory effect. BALB/c mice received 28 daily injections of PTH[1–34] (80 µg/kg/day) or L‐NAME (30 mg/kg/day), alone or in combination. Hindlimb blood perfusion was measured by laser Doppler imaging. Bone architecture, turnover and mechanical properties in the femur were analysed respectively by micro‐CT, histomorphometry and three‐point bending. PTH increased hindlimb blood flow by >30% within 10 min of injection (P < 0.001). Co‐treatment with L‐NAME blocked the action of PTH on blood flow, whereas L‐NAME alone had no effect. PTH treatment increased femoral cortical bone volume and formation rate by 20% and 110%, respectively (P < 0.001). PTH had no effect on trabecular bone volume in the femoral metaphysis although trabecular thickness and number were increased and decreased by 25%, respectively. Co‐treatment with L‐NAME restricted the PTH‐stimulated increase in cortical bone formation but had no clear‐cut effects in trabecular bone. Co‐treatment with L‐NAME did not affect the mechanical strength in femurs induced by iPTH. These results suggest that NO‐mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone. © 2016 The Authors. Cell Biochemistry and Function published by John Wiley & Sons, Ltd. 相似文献
8.
Trischitta F Denaro MG Faggio C 《Journal of experimental zoology. Part A, Comparative experimental biology》2004,301(1):49-62
Ion transport in the intestine of Gobius niger, a euryhaline teleost, was studied in both isotonic and hypotonic conditions. Isolated tissues, mounted in Ussing chambers and bilaterally perfused with isotonic Ringer solution, developed a serosa negative transepithelial voltage and a short circuit current indicating a net negative current in absorptive direction. Bilateral removal of Cl- and Na+ from the bathing solutions as well as the luminal removal of K+in the presence of Ba2+(10(-3) M) almost abolished both Vt and Isc. Similar results were obtained by adding bumetanide (10(-5)M) to the luminal bath while other inhibitors of Cl- transport mechanisms were ineffective. These observations suggest that salt absorption begins with a coupled entry of Na+, Cl-, and K+ across the apical membrane; a Ba2+inhibitable K+ conductance, demonstrated also by micropuncture experiments, recycles the ion into the lumen. Salt entry into the cell is driven by the operation of the basolateral Na+/K(+)-ATPase since serosal ouabain (10(-4)M) completely abolished both Vt and Isc; this pump also completes the Na(+) absorption. The inhibitory effect of both serosal bumetanide (10(-4)M) and SITS (5 x 10(-4)M) suggests that Cl- would leave the cell via the KCl cotransport, the Cl/HCO3- antiport and/or conductive pathways. Bilateral exposure of tissues to hypotonic media produced a reduction of both the transepithelial voltage and the short circuit current probably due to the activation of homeostatic ionic fluxes involved in cell volume regulation. The results of experiments with both isolated enterocytes and intestine exposed to hypotonic solution suggested that the recovery of cell volume, after the initial cell swelling, involves a parallel opening of K+ and Cl- channels to facilitate net solute and water effluxes from the cell. J. Exp. Zool. 301A:49-62, 2004. 相似文献
9.
Nuclear gene trees and the phylogenetic relationships of the mangabeys (Primates: Papionini) 总被引:3,自引:1,他引:2
Phylogenetic relationships of mangabeys within the Old World monkey tribe
Papionini are inferred from analyses of nuclear DNA sequences from five
unlinked loci. The following conclusions are strongly supported, based on
congruence among trees derived for the five separate gene regions: (1)
mangabeys are polyphyletic within the Papionini; (2) Cercocebus is the
sister taxon to the genus Mandrillus; and (3) Lophocebus belongs to a clade
with Papio and Theropithecus, with Papio as its most likely sister taxon.
Morphologically based phylogenies positing mangabey monophyly were
evaluated by mapping the sequences for each locus on these trees. The data
seem to fit these trees poorly in both maximum-parsimony and likelihood
analyses. Incongruence among nuclear gene trees occurred in the
interrelationships among Lophocebus, Papio, and Theropithecus. Several
factors that may account for this incongruence are discussed, including
sampling error, random lineage sorting, and introgression.
相似文献
10.
PPCMatrix: a PowerPC dotmatrix program to compare large genomic sequences against protein sequences 总被引:1,自引:0,他引:1
Summary : An interactive dotmatrix program for the MacOS was designed that
allows comparison of DNA to protein sequences using nested 3-frame
translations. Availability : Shareware, available at
http://copan.bioz.unibas.ch/software/ Contact : burglin@ubaclu. unibas.ch
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