全文获取类型
收费全文 | 1764篇 |
免费 | 248篇 |
出版年
2021年 | 25篇 |
2020年 | 29篇 |
2019年 | 30篇 |
2018年 | 27篇 |
2017年 | 27篇 |
2016年 | 28篇 |
2015年 | 57篇 |
2014年 | 61篇 |
2013年 | 82篇 |
2012年 | 89篇 |
2011年 | 81篇 |
2010年 | 61篇 |
2009年 | 42篇 |
2008年 | 73篇 |
2007年 | 79篇 |
2006年 | 51篇 |
2005年 | 58篇 |
2004年 | 52篇 |
2003年 | 47篇 |
2002年 | 48篇 |
2001年 | 49篇 |
2000年 | 51篇 |
1999年 | 54篇 |
1998年 | 25篇 |
1997年 | 24篇 |
1996年 | 23篇 |
1995年 | 27篇 |
1994年 | 26篇 |
1993年 | 22篇 |
1992年 | 35篇 |
1991年 | 44篇 |
1990年 | 34篇 |
1989年 | 44篇 |
1988年 | 30篇 |
1987年 | 37篇 |
1986年 | 34篇 |
1985年 | 38篇 |
1984年 | 40篇 |
1983年 | 25篇 |
1982年 | 27篇 |
1981年 | 13篇 |
1980年 | 23篇 |
1979年 | 23篇 |
1978年 | 13篇 |
1977年 | 20篇 |
1976年 | 10篇 |
1975年 | 15篇 |
1974年 | 26篇 |
1973年 | 23篇 |
1972年 | 11篇 |
排序方式: 共有2012条查询结果,搜索用时 15 毫秒
151.
Patients with LQT syndrome are prone to lifethreatening arrhythmias. After surviving such an event, implantation of an ICD is indicated. There are, however, special subtle demands in the treatment of these patients. In this case report we describe our findings in a patient with LQT1 syndrome, and the pitfalls that can and must be avoided. (Neth Heart J 2007;15:418-21.) 相似文献
152.
The redox-switch domain of Hsp33 functions as dual stress sensor 总被引:1,自引:0,他引:1
Ilbert M Horst J Ahrens S Winter J Graf PC Lilie H Jakob U 《Nature structural & molecular biology》2007,14(6):556-563
The redox-regulated chaperone Hsp33 is specifically activated upon exposure of cells to peroxide stress at elevated temperatures. Here we show that Hsp33 harbors two interdependent stress-sensing regions located in the C-terminal redox-switch domain of Hsp33: a zinc center sensing peroxide stress conditions and an adjacent linker region responding to unfolding conditions. Neither of these sensors works sufficiently in the absence of the other, making the simultaneous presence of both stress conditions a necessary requirement for Hsp33's full activation. Upon activation, Hsp33's redox-switch domain adopts a natively unfolded conformation, thereby exposing hydrophobic surfaces in its N-terminal substrate-binding domain. The specific activation of Hsp33 by the oxidative unfolding of its redox-switch domain makes this chaperone optimally suited to quickly respond to oxidative stress conditions that lead to protein unfolding. 相似文献
153.
Handisurya A Gilch S Winter D Shafti-Keramat S Maurer D Schätzl HM Kirnbauer R 《The FEBS journal》2007,274(7):1747-1758
Prion diseases are fatal neurodegenerative disorders caused by proteinaceous infectious pathogens termed prions (PrP(Sc)). To date, there is no prophylaxis or therapy available for these transmissible encephalopathies. Passive immunization with monclonal antibodies recognizing the normal host-encoded prion protein (PrP(C)) has been reported to abolish PrP(Sc) infectivity and to delay onset of disease. Because of established immunologic tolerance against the widely expressed PrP(C), active immunization appears to be difficult to achieve. To overcome this limitation, papillomavirus-like particles were generated that display a nine amino acid B-cell epitope, DWEDRYYRE, of the murine/rat prion protein in an immunogenic capsid surface loop, by insertion into the L1 major capsid protein of bovine papillomavirus type 1. The PrP peptide was selected on the basis of its previously suggested central role in prion pathogenesis. Immunization with PrP-virus-like particles induced high-titer antibodies to PrP in rabbit and in rat, without inducing overt adverse effects. As determined by peptide-specific ELISA, rabbit immune sera recognized the inserted murine/rat epitope and also cross-reacted with the homologous rabbit/human epitope differing in one amino acid residue. In contrast, rat immune sera recognized the murine/rat peptide only. Sera of both species reacted with PrP(C) in its native conformation in mouse brain and on rat pheochromocytoma cells, as determined by immunoprecipitation and fluorescence-activated cell sorting analysis. Importantly, rabbit anti-PrP serum contained high-affinity antibody that inhibited de novo synthesis of PrP(Sc) in prion-infected cells. If also effective in vivo, PrP-virus-like particle vaccination opens a unique possibility for immunologic prevention of currently fatal and incurable prion-mediated diseases. 相似文献
154.
155.
156.
We have characterized the temperature- and pressure-induced unfolding of staphylococcal nuclease (Snase) using high precision densitometric measurements. The changes in the apparent specific volume, expansion coefficient and isothermal compressibility were determined by these measurements. To our knowledge, these are the first measurements of the volume and isothermal compressibility changes of a protein undergoing pressure-induced unfolding. In order to aid in interpreting the temperature and pressure dependence of the apparent specific volume of Snase, we have also carried out differential scanning calorimetry under the solution conditions which are used for the volumetric studies. We have seen that large compensating volume and compressibility effects accompany the temperature and pressure-induced protein unfolding. Measurements of the apparent specific volume and thermal expansion coefficient of Snase at ambient pressure indicate the formation of a pre-transitional, molten globule type of intermediate structure about 10 degrees C below the actual unfolding temperature of the protein. Compared to the folded state, the apparent specific volume of the unfolded protein is about 0.3-0.5 % smaller. In addition, we investigated the pressure dependence of the apparent specific volume of Snase at a number of different temperatures. At 45 degrees C we calculate a decrease in apparent specific volume due to pressure-induced unfolding of -3.3 10(-3) cm(3) g(-1) or -55 cm(3) mol(-1). The threefold increase in compressibility between 40 and 70 MPa reflects a transition to a partially unfolded state, which is consistent with our results obtained for the radius of gyration of the pressure-denatured state of Snase. At the lower temperature of 35 degrees C, a significant increase in compressibility around 30 MPa is indicative of the formation of a pressure-induced molten globule-like intermediate. Changes in the apparent volume, expansion coefficient and isothermal compressibility are discussed in terms of instrinsic, hydrational and thermal contributions accompanying the unfolding transition. 相似文献
157.
Energetic cost of hovering flight in nectar-feeding bats (Phyllostomidae: Glossophaginae) and its scaling in moths, birds and bats 总被引:2,自引:0,他引:2
Voigt CC Winter Y 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1999,169(1):38-48
Three groups of specialist nectar-feeders covering a continuous size range from insects, birds and bats have evolved the
ability for hovering flight. Among birds and bats these groups generally comprise small species, suggesting a relationship
between hovering ability and size. In this study we established the scaling relationship of hovering power with body mass
for nectar-feeding glossophagine bats (Phyllostomidae). Employing both standard and fast-response respirometry, we determined
rates of gas exchange in Hylonycteris underwoodi (7 g) and Choeronycteris mexicana (13–18 g) during hover-feeding flights at an artificial flower that served as a respirometric mask to estimate metabolic
power input. The O2 uptake rate (V˙
o2) in ml g−1 h−1 (and derived power input) was 27.3 (1.12 W or 160 W kg−1) in 7-g Hylonycteris and 27.3 (2.63 W or 160 W kg−1) in 16.5-g Choeronycteris and thus consistent with measurements in 11.9-g Glossophagasoricina (158 W kg−1, Winter 1998). V˙
o2 at the onset of hovering was also used to estimate power during forward flight, because after a transition from level forward
to hovering flight gas exchange rates initially still reflect forward flight rates. V˙
o2 during short hovering events (<1.5 s) was 19.0 ml g−1 h−1 (1.8 W) in 16-g Choeronycteris, which was not significantly different from a previous, indirect estimate of the cost of level forward flight (2.1 W, Winter
and von Helversen 1998). Our estimates suggest that power input during hovering flight P
h
(W) increased with body mass M (kg) within 13–18-g Choeronycteris (n = 4) as P
h
= 3544 (±2057 SE) M
1.76 (±0.21 SE) and between different glossophagine bat species (n = 3) as P
h
= 128 (±2.4 SE) M
0.95 (±0.034 SE). The slopes of three scaling functions for flight power (hovering, level forward flight at intermediate speed and submaximal
flight power) indicate that: 1. The relationship between flight power to flight speed may change with body mass in the 6–30-g
bats from a J- towards a U-shaped curve. 2. A metabolic constraint (hovering flight power equal maximal flight power) may
influence the upper size limit of 30–35 g for this group of flower specialists.
Mass-specific power input (W kg−1) during hovering flight appeared constant with regard to body size (for the mass ranges considered), but differed significantly
(P < 0.001) between groups. Group means were 393 W kg−1 (sphingid moths), 261 W kg−1 (hummingbirds) and 159 W kg−1 (glossophagine bats). Thus, glossophagine bats expend the least metabolic power per unit of body mass supported during hovering
flight. At a metabolic power input of 1.1 W a glossophagine bat can generate the lift forces necessary for balancing 7 g against
gravitation, whereas a hummingbird can support 4 g and a sphingid moth only 3 g of body mass with the same amount of metabolic
energy. These differences in power input were not fully explained by differences in induced power output estimated from Rankine-Froude
momentum-jet theory.
Accepted: 10 November 1998 相似文献
158.
Phosphorylation of the amino terminus of maize sucrose synthase in relation to membrane association and enzyme activity 下载免费PDF全文
Sucrose synthase (SUS) is phosphorylated on a major, amino-terminal site located at Ser-15 (S15) in the maize (Zea mays) SUS1 protein. Site- and phospho-specific antibodies against a phosphorylated S15 (pS15) peptide allowed direct analysis of S15 phosphorylation in relation to membrane association. Immunoblots of the maize leaf elongation zone, divided into 4-cm segments, demonstrated that the abundance of soluble (s-SUS) and membrane (m-SUS) SUS protein showed distinct positional profiles. The content of m-SUS was maximal in the 4- to 8-cm segment where it represented 9% of total SUS and occurred as a peripheral membrane protein. In contrast, s-SUS was highest in the 12- to 16-cm segment. Relative to s-SUS, m-SUS was hypophosphorylated at S15 in the basal 4 cm but hyperphosphorylated in apical segments. Differing capabilities of the anti-pS15 and anti-S15 peptide antibodies to immunoprecipitate SUS suggested that phosphorylation of S15, or exposure of unphosphorylated SUS to slightly acidic pH, altered the structure of the amino terminus. These structural changes were generally coincident with the increased sucrose cleavage activity that occurs at pH values below 7.5. In vitro S15 phosphorylation of the S170A SUS protein by a maize calcium-dependent protein kinase (CDPK) significantly increased sucrose cleavage activity at low pH. Collectively, the results suggest that (1) SUS membrane binding is controlled in vivo; (2) relative pS15 content of m-SUS depends on the developmental state of the organ; and (3) phosphorylation of S15 affects amino-terminal conformation in a way that may stimulate the catalytic activity of SUS and influence membrane association. 相似文献
159.
Pedersen CB Bross P Winter VS Corydon TJ Bolund L Bartlett K Vockley J Gregersen N 《The Journal of biological chemistry》2003,278(48):47449-47458
Short chain acyl-CoA dehydrogenase (SCAD) deficiency is an inborn error of the mitochondrial fatty acid metabolism caused by rare variations as well as common susceptibility variations in the SCAD gene. Earlier studies have shown that a common variant SCAD protein (R147W) was impaired in folding, and preliminary experiments suggested that the variant protein displayed prolonged association with chaperonins and delayed formation of active enzyme. Accordingly, the molecular pathogenesis of SCAD deficiency may rely on intramitochondrial protein quality control mechanisms, including degradation and aggregation of variant SCAD proteins. In this study we investigated the processing of a set of disease-causing variant SCAD proteins (R22W, G68C, W153R, R359C, and Q341H) and two common variant proteins (R147W and G185S) that lead to reduced SCAD activity. All SCAD proteins, including the wild type, associate with mitochondrial hsp60 chaperonins; however, the variant SCAD proteins remained associated with hsp60 for prolonged periods of time. Biogenesis experiments at two temperatures revealed that some of the variant proteins (R22W, G68C, W153R, and R359C) caused severe misfolding, whereas others (R147W, G185S, and Q341H) exhibited a less severe temperature-sensitive folding defect. Based on the magnitude of in vitro defects, these SCAD proteins are characterized as folding-defective variants and mild folding variants, respectively. Pulse-chase experiments demonstrated that the variant SCAD proteins either triggered proteolytic degradation by mitochondrial proteases or, especially at elevated temperature, aggregation of non-native conformers. The latter finding may indicate that accumulation of aggregated SCAD proteins may play a role in the pathogenesis of SCAD deficiency. 相似文献
160.
Schäffner J Winter J Rudolph R Schwarz E 《Applied and environmental microbiology》2001,67(9):3994-4000
Attempts were made to engineer the periplasm of Escherichia coli to an expression compartment of heterologous proteins in their native conformation. As a first approach the low-molecular-size additive L-arginine and the redox compound glutathione (GSH) were added to the culture medium. Addition of 0.4 M L-arginine and 5 mM reduced GSH increased the yield of a native tissue-type plasminogen activator variant (rPA), consisting of the kringle-2 and the protease domain, and a single-chain antibody fragment (scFv) up to 10- and 37-fold, respectively. A variety of other medium additives also had positive effects on the yield of rPA. In a second set of experiments, the effects of cosecreted ATP-independent molecular chaperones on the yields of native therapeutic proteins were investigated. At optimized conditions, cosecretion of E. coli DnaJ or murine Hsp25 increased the yield of native rPA by a factor of 170 and 125, respectively. Cosecretion of DnaJ also dramatically increased the amount of a second model protein, native proinsulin, in the periplasm. The results of this study are anticipated to initiate a series of new approaches to increase the yields of native, disulfide-bridged, recombinant proteins in the periplasm of E. coli. 相似文献