全文获取类型
收费全文 | 101篇 |
免费 | 11篇 |
专业分类
112篇 |
出版年
2022年 | 1篇 |
2021年 | 2篇 |
2019年 | 1篇 |
2018年 | 3篇 |
2017年 | 1篇 |
2016年 | 4篇 |
2015年 | 3篇 |
2014年 | 6篇 |
2013年 | 4篇 |
2012年 | 11篇 |
2011年 | 7篇 |
2010年 | 3篇 |
2009年 | 8篇 |
2008年 | 4篇 |
2007年 | 9篇 |
2006年 | 5篇 |
2005年 | 7篇 |
2004年 | 5篇 |
2003年 | 3篇 |
2002年 | 2篇 |
2001年 | 6篇 |
2000年 | 2篇 |
1999年 | 2篇 |
1996年 | 1篇 |
1991年 | 1篇 |
1990年 | 3篇 |
1989年 | 2篇 |
1987年 | 1篇 |
1979年 | 1篇 |
1978年 | 1篇 |
1975年 | 3篇 |
排序方式: 共有112条查询结果,搜索用时 12 毫秒
1.
2.
The total synthesis of two ring-homo prostaglandin analogues XIII and XV, from a common intermediate IV is described. 相似文献
3.
A global increase in invasive infections due to group A Streptococcus (S. pyogenes or GAS) has been observed since the 1980s, associated with emergence of a clonal group of strains of the M1T1 serotype. Among other virulence attributes, the M1T1 clone secretes NAD+-glycohydrolase (NADase). When GAS binds to epithelial cells in vitro, NADase is translocated into the cytosol in a process mediated by streptolysin O (SLO), and expression of these two toxins is associated with enhanced GAS intracellular survival. Because SLO is required for NADase translocation, it has been difficult to distinguish pathogenic effects of NADase from those of SLO. To resolve the effects of the two proteins, we made use of anthrax toxin as an alternative means to deliver NADase to host cells, independently of SLO. We developed a novel method for purification of enzymatically active NADase fused to an amino-terminal fragment of anthrax toxin lethal factor (LFn-NADase) that exploits the avid, reversible binding of NADase to its endogenous inhibitor. LFn-NADase was translocated across a synthetic lipid bilayer in vitro in the presence of anthrax toxin protective antigen in a pH-dependent manner. Exposure of human oropharyngeal keratinocytes to LFn-NADase in the presence of protective antigen resulted in cytosolic delivery of NADase activity, inhibition of protein synthesis, and cell death, whereas a similar construct of an enzymatically inactive point mutant had no effect. Anthrax toxin-mediated delivery of NADase in an amount comparable to that observed during in vitro infection with live GAS rescued the defective intracellular survival of NADase-deficient GAS and increased the survival of SLO-deficient GAS. Confocal microscopy demonstrated that delivery of LFn-NADase prevented intracellular trafficking of NADase-deficient GAS to lysosomes. We conclude that NADase mediates cytotoxicity and promotes intracellular survival of GAS in host cells. 相似文献
4.
Felicori LF Souza CT Velarde DT Magalhaes A Almeida AP Figueiredo S Richardson M Diniz CR Sanchez EF 《Protein expression and purification》2003,30(1):32-42
A kallikrein-like proteinase of Lachesis muta muta (bushmaster) venom, designated LV-Ka, was purified by gel filtration and anion exchange chromatographies. Physicochemical studies indicated that the purified enzyme is a 33 kDa monomeric glycoprotein, the Mr of which fell to 28 kDa after deglycosylation with PNGase F. Approximately 77% of the protein sequence was determined by sequencing the various fragments derived from digestions with endoproteases. The partial sequence obtained suggests that LV-Ka is of a similar size to other serine proteinases (i.e., approximately 234 amino acid residues). Sequence studies on the NH2-terminal region of the protein indicate that LV-Ka shares a high degree of sequence homology with the kallikrein-like enzymes EI and EII from Crotalus atrox, with crotalase from Crotalus adamanteus and significant homology with other serine proteinases from snake venoms and vertebrate serum enzymes. LV-Ka showed kallikrein-like activity, releasing bradikinin from kininogen as evidenced by guinea pig bioassay. In addition, intravenous injection of the proteinase (0.8 microg/g) was shown to lower blood pressure in experimental rats. In vitro, the isolated proteinase was shown to have neither fibrin(ogeno)lytic activity nor coagulant effect. LV-Ka was active upon the kallikrein substrates S-2266 and S-2302 (specific activity=13.0 and 31.5 U/mg, respectively; crude venom=0.25 and 6.0 U/mg) but had no proteolytic effect on dimethylcasein and insulin B chain. Its enzymatic activity was inhibited by NPGB and PMSF, indicating that the enzyme is a serine proteinase. Interestingly, one of the other reactions catalyzed by plasma kallikrein, the activation of plasminogen was one of the activities exhibited by LV-Ka. 相似文献
5.
6.
Stimac G Dimanovski J Trnski D Katusić J Ruzić B Spajić B Reljić A Padovan M Kraus O 《Collegium antropologicum》2007,31(4):1055-1060
We demonstrate the evolution of the clinical presentation and outcomes for patients with clinically localized prostate cancer (PC) treated with radical retropubic prostatectomy (RRP) at our department, emphasizing epidemiologic significance of changes during the 10-year period. We assessed the annual trends for changes in patients age, preoperative prostate specific antigen (PSA), preoperative versus postoperative stages and Gleason grades, organ confined status and surgical margin status. A total of 488 RRPs were performed from January 1996 to December 2005 with the annual frequency increased from 8 to 129 (1512.5%). Mean patient age increased from 61.5 to 66.12 years in 2005, with the percentage of men aged more than 70 years increased from 12.5 to 26.5%, respectively. The detection of PC based solely on pathological PSA levels (as indication for prostate biopsy) rose impressively from 25.5 to 70% and the rates of postoperative organ-confined disease also increased significantly from 25 to 74.7%. Mean preoperative PSA decreased from 16.7 to 9.89 ng/mL. On the contrary, there was an increase in percentage of patients with preoperative PSA values ranging from 4 to 10 ng/mL (from 20 to 65.4%). Positive surgical margin rate decreased from 49.4 to 25% and percent of patients receiving neoadjuvant therapy decreased from 78.5 to 5.4%. Proportion of patients who were undergraded decreased from 75.1 to 31.7%. The rates of understaging have remained relatively stable over the years. During the study period, PC was increasingly detected by prostate biopsy on the basis of a pathological PSA level only and shifted significantly to more organ-confined stages with more favourable outcomes for pathological variables due to a more accurate assessment of clinical stage prior to surgery, reduced use of neoadjuvant therapy and improved surgical technique. Our data also argue strongly that routine PSA testing should be expanded and not restricted. 相似文献
7.
Melisa Kiyamu Gianpietro Elías Fabiola León‐Velarde María Rivera‐Chira Tom D. Brutsaert 《American journal of physical anthropology》2015,156(3):363-373
High altitude natives are reported to have outstanding work capacity in spite of the challenge of oxygen transport and delivery in hypoxia. To evaluate the developmental effect of lifelong exposure to hypoxia on aerobic capacity, VO2peak was measured on two groups of Peruvian Quechua subjects (18–35 years), who differed in their developmental exposure to altitude. Male and female volunteers were recruited in Lima, Peru (150 m), and were divided in two groups, based on their developmental exposure to hypoxia, those: a) Born at sea‐level individuals (BSL), with no developmental exposure to hypoxia (n = 34) and b) Born at high‐altitude individuals (BHA) with full developmental exposure to hypoxia (n = 32), but who migrated to sea‐level as adults (>16‐years‐old). Tests were conducted both in normoxia (BP = 750 mm Hg) and normobaric hypoxia at sea‐level (BP = 750 mm Hg, FiO2 = 0.12, equivalent to 4,449 m), after a 2‐month training period (in order to control for initial differences in physical fitness) at sea‐level. BHA had a significantly higher VO2peak at hypoxia (40.31 ± 1.0 ml/min/kg) as compared to BSL (35.78 ± 0.96 ml/min/kg, P = 0.001), adjusting for sex. The decrease of VO2peak at HA relative to SL (ΔVO2peak) was not different between groups, controlling for baseline levels (VO2peak at sea‐level) and sex (BHA = 0.35 ± 0.04 l/min, BSL = 0.44 ± 0.04 l/min; P = 0.12). Forced vital capacity (controlling for height) and the residuals of VO2peak (controlling for weight) had a significant association in the BHA group only (r = 0.155; P = 0.031). In sum, results indicate that developmental exposure to altitude constitutes an important factor to determine superior exercise performance. Am J Phys Anthropol 156:363–373, 2015. © 2014 Wiley Periodicals, Inc. 相似文献
8.
Gustavo Ciudad Isaac Reyes Milko A. Jorquera Laura Azócar Lukas Y. Wick Rodrigo Navia 《World journal of microbiology & biotechnology》2011,27(11):2505-2512
A novel three-phase solid–gas–liquid bioreactor (SGLB) concept using gaseous alcohol and liquid rapeseed oil with immersed
microorganisms overlying a nutrient agar phase (solid) is proposed for biodiesel (fatty acid alkyl esters, FAAE) production
based on the high hydrophobicity and negative surface charge showed by the fungi Rhizopus oryzae. This novel bioreactor was thought to increase oil bioavailability and decrease alcohol toxicity for effective microbial
growth, reaching high yields of FAAE production without any pretreatment. High growth rates were reached for R. oryzae using a SGLB simultaneously reaching a high FAAE production yield, up to 50% using methanol and up to 70% using ethanol at
144 h of incubation at 20°C. To compare the effect of gaseous alcohol, the same experiments were carried out in a three-phase
solid–liquid–liquid bioreactor (SLLB), where the alcohol was added in liquid phase, showing significant R. oryzae growth but no FAAE formation. This suggests that the inhibitory effect of alcohol is more significant in lipase activity
than in R. oryzae growth, and the use of alcohol in gaseous phase may decrease both of them. The experimental procedure using SGLB showed that
when R. oryzae is maintained alive, it can catalyze at the same time the hydrolysis, esterification and transesterification of triglycerides
from rapeseed oil, but its activity strongly depends on the used growth media. Therefore, the application of gaseous alcohol
coupled with R. oryzae as immobilized whole cell catalysts may be a potential alternative to the use of commercial lipases for biodiesel production. 相似文献
9.
Ciudad G Reyes I Azócar L Briones R Jorquera M Wick LY Navia R 《New biotechnology》2011,28(4):375-381
The high cost of commercial lipases limits their industrial application in the production of biodiesel or fatty acid methyl esters (FAME). This disadvantage has encouraged the search for lipase-producing microorganisms (LPMs) as potential whole cell catalysts for FAME production. The aim of this study, therefore, was to evaluate innovative procedures for easy selection and testing of LPMs as a low-cost whole cell catalyst, based on catalytic performance, methanol tolerance and physico-chemical cell surface properties. The latter (in particular the cell surface hydrophobicity and charge) were analyzed because of their crucial role in microbial adhesion to surfaces and the concomitant increase in cell immobilization and bioavailability of hydrophobic substrates. Biocatalysis experiments performed in the presence of nutrient, rapeseed oil and methanol were an effective tool for studying and identifying, in just two experiments, the capacity of different LPMs as biocatalysts in organic media, as well as the methanol tolerance of the cell and the lipase. This indicates the potential for using live microorganisms for FAME production. Another finding was that the inhibitory effect of methanol is more significant for lipase activity than LPM growth, indicating that the way in which alcohol is supplied to the reaction is a crucial step in FAME production by biocatalysts. According to these results, the application of these innovative assessments should simplify the search for new strains which are able to effectively catalyze the FAME production process. 相似文献
10.
Luaces I Aguirre E García-Montijano M Velarde J Tesouro MA Sánchez C Galka M Fernández P Sainz A 《Journal of wildlife diseases》2005,41(4):810-815
A wild injured Iberian lynx (Lynx pardinus) was taken from the Sierra Morena population. During the health check small intraerythrocytic piroplasms, morphologically indistinguishable from other feline piroplasms, were observed in Wright-Giemsa-stained blood films. Amplification by polymerase chain reaction of a portion of the 18S nuclear small subunit (NSS) rRNA gene and sequencing revealed similarity of the unknown organism with sequences obtained from Pallas's cat from Mongolia and from a domestic cat in Spain. In a retrospective (1993-2003) study of 50 Iberian lynx tissue samples, no amplifications of the 18S NSS rRNA gene of the organism were obtained. This is the first report of a naturally occurring erythroparasitemia in the Iberian lynx and the first documented case of naturally occurring piroplasm infection in a free-ranging felid from Europe. 相似文献