全文获取类型
收费全文 | 243篇 |
免费 | 37篇 |
专业分类
280篇 |
出版年
2022年 | 4篇 |
2017年 | 2篇 |
2015年 | 8篇 |
2014年 | 9篇 |
2013年 | 6篇 |
2012年 | 8篇 |
2011年 | 4篇 |
2010年 | 8篇 |
2009年 | 7篇 |
2008年 | 7篇 |
2007年 | 8篇 |
2006年 | 6篇 |
2005年 | 9篇 |
2004年 | 11篇 |
2003年 | 7篇 |
2002年 | 9篇 |
2001年 | 15篇 |
2000年 | 10篇 |
1999年 | 11篇 |
1998年 | 4篇 |
1997年 | 4篇 |
1995年 | 3篇 |
1994年 | 5篇 |
1993年 | 4篇 |
1992年 | 6篇 |
1991年 | 6篇 |
1990年 | 10篇 |
1989年 | 3篇 |
1988年 | 4篇 |
1987年 | 3篇 |
1982年 | 2篇 |
1980年 | 2篇 |
1979年 | 6篇 |
1977年 | 3篇 |
1976年 | 4篇 |
1975年 | 4篇 |
1974年 | 7篇 |
1973年 | 3篇 |
1972年 | 5篇 |
1971年 | 2篇 |
1970年 | 2篇 |
1969年 | 5篇 |
1968年 | 4篇 |
1967年 | 3篇 |
1966年 | 2篇 |
1965年 | 3篇 |
1964年 | 3篇 |
1961年 | 2篇 |
1946年 | 2篇 |
1941年 | 1篇 |
排序方式: 共有280条查询结果,搜索用时 15 毫秒
71.
Wolfgang Sperl Daniela Skladal Erich Gnaiger Markus Wyss Udo Mayr Josef Hager Frank Norbert Gellerich 《Molecular and cellular biochemistry》1997,174(1-2):71-78
High resolution respirometry in combination with the skinned fiber technique offers the possibility to study mitochondrial function routinely in small amounts of human muscle. During a period of 2 years, we investigated mitochondrial function in skeletal muscle tissue of 13 patients (average age = 5.8 years). In all of them, an open muscle biopsy was performed for diagnosis of their neuromuscular disorder. Mitochondrial oxidation rates were measured with a highly sensitive respirometer. Multiple substrate-inhibitor titration was applied for investigation of mitochondrial function. About 50 mg fibers were sufficient to obtain maximal respiratory rates for seven different substrates (pyruvate/malate, glutamate/malate, octanoylcarnitine/malate, palmitoylcarnitine /malate, succinate, durochinol and ascorbate/TMPD). Decreased respiration rates with reference to the wet weight of the permeabilized fiber could immediately be detected during the course of measurements.In 4 patients with mitochondrial encephalomyopathy (MEM) the respiration pattern indicated a specific mitochondrial enzyme defect, which was confirmed in every patient by measurements of the individual enzymes (one patient with PDHC deficiency, one with complex I deficiency and two patients with combined complex I and IV deficiency). In the 6 patients with spinal muscular atrophy (SMA) oxidation rates were found to be decreased to 23 ± 5% of controls. The normalized respiration pattern was comparable to that of the controls indicating a decreased content of mitochondria in SMA muscle with normal functional properties. Also in the 3 patients with Duchenne muscular dystrophy (DMD) decreased oxidation rates (42 ± 5%) were detected. In addition a low RCI (1.2) indicated a loose coupling of oxidative phosphorylation in the mitochondria of these patients.It is concluded that investigation of mitochondrial function in saponin skinned muscle fibers using high resolution respirometry in combination with multiple substrate titration offers a valuable tool for evaluation of mitochondrial alterations in muscle biopsies of children suffering from neuromuscular disorders. (Mol Cell Biochem 174: 71–78, 1997) 相似文献
72.
C Wyss 《Somatic cell genetics》1979,5(1):23-28
Yeast extract, a component of Drosophila cell culture media, is shown to contain substances of high, intermediate, and low molecular weight, that are, respectively, essential, inhibitory, and stimulatory for colony formation in semisolid agar medium. Furthermore, it is shown that high concentrations of pyridoxal greatly increase the cloning efficiency of Drosophilia cells. A cloning method with line Kc is described which routinely gives cloning efficiencies in excess of 20%. 相似文献
73.
Reconstitution of active octameric mitochondrial creatine kinase from two genetically engineered fragments. 总被引:3,自引:1,他引:2
M. Gross M. Wyss E. M. Furter-Graves T. Wallimann R. Furter 《Protein science : a publication of the Protein Society》1996,5(2):320-330
Creatine kinase (CK) has been postulated to consist of two flexibly hinged domains. A previously demonstrated protease-sensitive site in M-CK (Morris & Jackson, 1991) has directed our attempts to dissect mitochondrial CK (Mi-CK) into two protein fragments encompassing amino acids [1-167] and [168-380]. When expressed separately in Escherichia coli, the two fragments yielded large amounts of insoluble inclusion bodies, from which the respective polypeptides could be purified by a simple two-step procedure. In contrast, co-expression of the two fragments yielded a soluble, active, and correctly oligomerizing enzyme. This discontinuous CK showed nearly full specific activity and was virtually indistinguishable from native Mi-CK by far- and near-UV CD. However, the positive cooperativity of substrate binding was abolished, suggesting a role of the covalent domain linkage in the crosstalk between the substrate binding sites for ATP and creatine. The isolated C-terminal fragment refolded into a native-like conformation in vitro, whereas the N-terminal fragment was largely unfolded. Prefolded [168-380] interacted in vitro with [1-167] to form an active enzyme. Kinetic analysis indicated that the fragments associate rapidly and with high affinity (1/K1 = 17 microM) and then isomerize slowly to an active enzyme (k2 = 0.12 min-1; k-2 = 0.03 min-1). Our data suggest that the C-terminal fragment of Mi-CK represents an autonomous folding unit, and that the folding of the C-terminal part might precede the conformational stabilization of the N-terminal moiety in vivo. 相似文献
74.
75.
Jean-Philippe Surivet Roland Lange Christian Hubschwerlen Wolfgang Keck Jean-Luc Specklin Daniel Ritz Daniel Bur Hans Locher Peter Seiler Daniel Stefan Strasser Lars Prade Christopher Kohl Christine Schmitt Gaëlle Chapoux Eser Ilhan Nadia Ekambaram Alcibiade Athanasiou Andreja Knezevic Daniela Sabato Alain Chambovey Pierre Wyss 《Bioorganic & medicinal chemistry letters》2012,22(21):6705-6711
A series of 2-amino-[1,8]-naphthyridine-3-carboxamides (ANCs) with potent inhibition of bacterial NAD+-dependent DNA ligases (LigAs) evolved from a 2,4-diaminopteridine derivative discovered by HTS. The design was guided by several highly resolved X-ray structures of our inhibitors in complex with either Streptococcus pneumoniae or Escherichia coli LigA. The structure–activity-relationship based on the ANC scaffold is discussed. The in-depth characterization of 2-amino-6-bromo-7-(trifluoromethyl)-[1,8]-naphthyridine-3-carboxamide, which displayed promising in vitro (MIC Staphylococcus aureus 1 mg/L) and in vivo anti-staphylococcal activity, is presented. 相似文献
76.
77.
Barbara D'haene Catia Attanasio Diane Beysen Jose Dostie Edmond Lemire Philippe Bouchard Michael Field Kristie Jones Birgit Lorenz Bjrn Menten Karen Buysse Filip Pattyn Marc Friedli Catherine Ucla Colette Rossier Carine Wyss Frank Speleman Anne De Paepe Job Dekker Stylianos E. Antonarakis Elfride De Baere 《PLoS genetics》2009,5(6)
To date, the contribution of disrupted potentially cis-regulatory conserved non-coding sequences (CNCs) to human disease is most likely underestimated, as no systematic screens for putative deleterious variations in CNCs have been conducted. As a model for monogenic disease we studied the involvement of genetic changes of CNCs in the cis-regulatory domain of FOXL2 in blepharophimosis syndrome (BPES). Fifty-seven molecularly unsolved BPES patients underwent high-resolution copy number screening and targeted sequencing of CNCs. Apart from three larger distant deletions, a de novo deletion as small as 7.4 kb was found at 283 kb 5′ to FOXL2. The deletion appeared to be triggered by an H-DNA-induced double-stranded break (DSB). In addition, it disrupts a novel long non-coding RNA (ncRNA) PISRT1 and 8 CNCs. The regulatory potential of the deleted CNCs was substantiated by in vitro luciferase assays. Interestingly, Chromosome Conformation Capture (3C) of a 625 kb region surrounding FOXL2 in expressing cellular systems revealed physical interactions of three upstream fragments and the FOXL2 core promoter. Importantly, one of these contains the 7.4 kb deleted fragment. Overall, this study revealed the smallest distant deletion causing monogenic disease and impacts upon the concept of mutation screening in human disease and developmental disorders in particular. 相似文献
78.
Lijun Dai Geeta Datta Zhenghao Zhang Himanshu Gupta Rakesh Patel Jaideep Honavar Sarika Modi J. Michael Wyss Mayakonda Palgunachari G. M. Anantharamaiah C. Roger White 《Journal of lipid research》2010,51(9):2695-2705
High density lipoprotein (HDL) and apolipoprotein A-I (apoA-I) reduce inflammatory responses to lipopolysaccharide (LPS). We tested the hypothesis that the apoA-I mimetic peptide 4F prevents LPS-induced defects in blood pressure and vascular reactivity. Systolic blood pressure (SBP) was measured in rats at baseline and 6 h after injection of LPS (10 mg/kg) or saline vehicle. Subgroups of LPS-treated rats also received 4F (10 mg/kg) or scrambled 4F (Sc-4F). LPS administration reduced SBP by 35% compared with baseline. 4F attenuated the reduction in SBP in LPS-treated rats (17% reduction), while Sc-4F was without effect. Ex vivo studies showed a reduced contractile response to phenylephrine (PE) in aortae of LPS-treated rats (ED50 = 459 ± 83 nM) compared with controls (ED50 = 57 ± 6 nM). This was associated with nitric oxide synthase 2 (NOS2) upregulation. 4F administration improved vascular contractility (ED50 = 60 ± 9 nM), reduced aortic NOS2 protein, normalized plasma levels of NO metabolites, and reduced mortality in LPS-treated rats. These changes were associated with a reduction in plasma endotoxin activity. In vivo administration of 14C-4F and Bodipy-LPS resulted in their colocalization and retention in the HDL fraction. It is proposed that 4F promotes the localization of LPS to the HDL fraction, resulting in endotoxin neutralization. 4F may thus prevent LPS-induced hemodynamic changes associated with NOS2 induction. 相似文献
79.
A novel chemistry has been developed for the production of capsules composed of a hydrophobic liquid core surrounded by a cross-linked polyacrylamide/alginate membrane. These liquid-core capsules may be used in capsular perstraction for the removal of inhibitory products from bioprocesses and bioconversions. They have the advantage of having a high surface area to promote rapid mass transfer, while separation of the organic core phase from the aqueous environment by the capsule membrane prevents the formation of stable emulsions and potential problems associated with toxicity of the organic phase for microbial cells or enzymes. Monodisperse spherical liquid-core capsules of between 800 microm and 1.6 mm diameter, with high mechanical resistance, have been prepared by co-extrusion, using the jet break-up technique. Capsules produced from a solution of MBA/total monomer (5%) were found to be more elastic and have a higher burst force when exposed to chelating agents such as phosphate or citrate. The mechanical resistance was unaffected by buffer solutions in the pH range 4-9 and after sterilization at 121 degrees C for 20 min. Capsules having membranes composed of a copolymer of acrylamide and N-hydroxymethylacrylamide exhibited even higher mechanical stability toward chelating agents. 相似文献
80.
Two paralogous families of a two-gene subtilisin operon are widely distributed in oral treponemes 下载免费PDF全文
Correia FF Plummer AR Ellen RP Wyss C Boches SK Galvin JL Paster BJ Dewhirst FE 《Journal of bacteriology》2003,185(23):6860-6869
Certain oral treponemes express a highly proteolytic phenotype and have been associated with periodontal diseases. The periodontal pathogen Treponema denticola produces dentilisin, a serine protease of the subtilisin family. The two-gene operon prcA-prtP is required for expression of active dentilisin (PrtP), a putative lipoprotein attached to the treponeme's outer membrane or sheath. The purpose of this study was to examine the diversity and structure of treponemal subtilisin-like proteases in order to better understand their distribution and function. The complete sequences of five prcA-prtP operons were determined for Treponema lecithinolyticum, "Treponema vincentii," and two canine species. Partial operon sequences were obtained for T. socranskii subsp. 04 as well as 450- to 1,000-base fragments of prtP genes from four additional treponeme strains. Phylogenetic analysis demonstrated that the sequences fall into two paralogous families. The first family includes the sequence from T. denticola. Treponemes possessing this operon family express chymotrypsin-like protease activity and can cleave the substrate N-succinyl-alanyl-alanyl-prolyl-phenylalanine-p-nitroanilide (SAAPFNA). Treponemes possessing the second paralog family do not possess chymotrypsin-like activity or cleave SAAPFNA. Despite examination of a range of protein and peptide substrates, the specificity of the second protease family remains unknown. Each of the fully sequenced prcA and prtP genes contains a 5' hydrophobic leader sequence with a treponeme lipobox. The two paralogous families of treponeme subtilisins represent a new subgroup within the subtilisin family of proteases and are the only subtilisin lipoprotein family. The present study demonstrated that the subtilisin paralogs comprising a two-gene operon are widely distributed among treponemes. 相似文献