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991.
992.
993.
Single-nucleotide variations are the most widely distributed genetic markers in the human genome. A subset of these variations, the substitution mutations, are responsible for most genetic disorders. As single nucleotide polymorphism (SNP) markers are being developed for molecular diagnosis of genetic disorders and large-scale population studies for genetic analysis of complex traits, a simple, sensitive, and specific test for single nucleotide changes is highly desirable. In this report we describe the development of a homogeneous DNA detection method that requires no further manipulations after the initial reaction is set up. This assay, named dye-labeled oligonucleotide ligation (DOL), combines the PCR and the oligonucleotide ligation reaction in a two-stage thermal cycling sequence with fluorescence resonance energy transfer (FRET) detection monitored in real time. Because FRET occurs only when the donor and acceptor dyes are in close proximity, one can infer the genotype or mutational status of a DNA sample by monitoring the specific ligation of dye-labeled oligonucleotide probes. We have successfully applied the DOL assay to genotype 10 SNPs or mutations. By designing the PCR primers and ligation probes in a consistent manner, multiple assays can be done under the same thermal cycling conditions. The standardized design and execution of the DOL assay means that it can be automated for high-throughput genotyping in large-scale population studies. 相似文献
994.
995.
Thrombocytopaenia, or platelet aggregation, is a serious complication of African trypanosomiasis. The biochemical basis is
not clearly known. Proteases are known potent inducers of blood coagulation and platelet aggregation, and unknown factors
released by Trypanosoma brucei have been shown to induce platelet aggregation. In attempts to define the biochemical mechanisms involved in thrombocytopaenia
we purified and characterised a major proteolytic enzyme released extracellularly by T. brucei. Actively motile trypanosomes released proteins into the medium (phosphate/saline/glucose, pH 8.0) in which the organisms
were incubated in vitro. The Mr of the released polypeptides ranged from 15 to >200 kDa, amongst which are proteases. One of the major protein bands, a 250 kDa
protease, was purified to homogeneity by ammonium sulfate precipitation followed by diethylaminoethyl (DEAE)-cellulose chromatography
and Sephacryl S-300 gel filtration. The protease migrated as a single band of 63 kDa upon electrophoresis in both denaturing
and non-denaturing gel co-polymerised with gelatin. The enzyme was strongly active against Z-ARR-AFC peptide substrate, with
a pH optimum of 7.0. The proteolytic activity was enhanced by dithiothreitol and inhibited by E-64, leupeptin, TPCK and antipain.
The released proteolytic enzyme is putatively identified as a cathepsin B-like cysteine protease.
Received: 18 May 1998 / Accepted: 30 September 1998 相似文献
996.
The Mechanism of Renal Glomerular Capillary Retention of Carbon in Disseminated Intravascular Coagulation 下载免费PDF全文
Kenneth J. Donald Alan N. Whitaker Ian H. Bunce 《The American journal of pathology》1973,70(2):245-252
In rabbits, when renal glomerular capillary thrombosis was induced by an infusion of thrombin, injected colloidal carbon particles localized within glomeruli. When administered after the thrombin had been infused, carbon was attached to the surface of fibrin deposits. Carbon also attached to platelets within capillaries containing fibrin, but not to leukocytes or endothelial cells. If administered simultaneously with thrombin, the carbon was incorporated into the developing mass of fibrin. These observations demonstrate that disseminated intravascular coagulation leads to the deposition of colloidal materials from the circulating blood in regions remote from reticuloendothelial organs, and illustrate a nonimmunologic mechanism whereby foreign particles and abnormal proteins might be trapped within the renal microcirculation. 相似文献
997.
Madhavan S Weber KA Stinear JW 《Experimental brain research. Experimentelle Hirnforschung. Expérimentation cérébrale》2011,209(1):9-17
We set out to answer two questions with this study: 1. Can stroke patients improve voluntary control of their paretic ankle by practising a visuo-motor ankle-tracking task? 2. Are practice effects enhanced with non-invasive brain stimulation? A carefully selected sample of chronic stroke patients able to perform the experimental task attended three data collection sessions. Facilitatory transcranial direct current stimulation (tDCS) was applied in a random order over the lower limb primary motor cortex of the lesioned hemisphere or the non-lesioned hemisphere or sham stimulation was delivered over the lesioned hemisphere. In each session, tDCS was applied as patients practiced tracking a sinusoidal waveform for 15 min using dorsiflexion–plantarflexion movements of their paretic ankle. The difference in tracking error prior to, and after, the 15 min of practice was calculated. A practice effect was revealed following sham stimulation, and this effect was enhanced with tDCS applied over the lesioned hemisphere. The practice effect observed following sham stimulation was eliminated by tDCS applied over the non-lesioned hemisphere. The study provides the first evidence that non-invasive brain stimulation applied to the lesioned motor cortex of moderate- to well-recovered stroke patients enhances voluntary control of the paretic ankle. The results provide a basis for examining whether this enhanced ankle control can be induced in patients with greater impairments and whether enhanced control of a single or multiple lower limb joints improves hemiparetic gait patterns. 相似文献
998.
Guida Landouré Peng‐Peng Zhu Charles M. Lourenço Janel O. Johnson Camilo Toro Katherine V. Bricceno Carlo Rinaldi Katherine G. Meilleur Modibo Sangaré Oumarou Diallo Tyler M. Pierson Hiroyuki Ishiura Shoji Tsuji Nichole Hein John K. Fink Marion Stoll Garth Nicholson Michael A. Gonzalez Fiorella Speziani Alexandra Dürr Giovanni Stevanin Leslie G. Biesecker for the NIH Intramural Sequencing Center John Accardi Dennis M. D. Landis William A. Gahl Bryan J. Traynor Wilson Marques Jr Stephan Züchner Craig Blackstone Kenneth H. Fischbeck Barrington G. Burnett 《Human mutation》2013,34(10):1357-1360
We report here the genetic basis for a form of progressive hereditary spastic paraplegia (SPG43) previously described in two Malian sisters. Exome sequencing revealed a homozygous missense variant (c.187G>C; p.Ala63Pro) in C19orf12, a gene recently implicated in neurodegeneration with brain iron accumulation (NBIA). The same mutation was subsequently also found in a Brazilian family with features of NBIA, and we identified another NBIA patient with a three‐nucleotide deletion (c.197_199del; p.Gly66del). Haplotype analysis revealed that the p.Ala63Pro mutations have a common origin, but MRI scans showed no brain iron deposition in the Malian SPG43 subjects. Heterologous expression of these SPG43 and NBIA variants resulted in similar alterations in the subcellular distribution of C19orf12. The SPG43 and NBIA variants reported here as well as the most common C19orf12 missense mutation reported in NBIA patients are found within a highly conserved, extended hydrophobic domain in C19orf12, underscoring the functional importance of this domain. 相似文献
999.
1000.
Maisel A Mueller C Adams K Anker SD Aspromonte N Cleland JG Cohen-Solal A Dahlstrom U DeMaria A Di Somma S Filippatos GS Fonarow GC Jourdain P Komajda M Liu PP McDonagh T McDonald K Mebazaa A Nieminen MS Peacock WF Tubaro M Valle R Vanderhyden M Yancy CW Zannad F Braunwald E 《European journal of heart failure》2008,10(9):824-839
Natriuretic peptide (NP) levels (B-type natriuretic peptide (BNP) and N-terminal proBNP) are now widely used in clinical practice and cardiovascular research throughout the world and have been incorporated into most national and international cardiovascular guidelines for heart failure. The role of NP levels in state-of-the-art clinical practice is evolving rapidly. This paper reviews and highlights ten key messages to clinicians: 相似文献