全文获取类型
收费全文 | 309篇 |
免费 | 25篇 |
出版年
2021年 | 2篇 |
2020年 | 2篇 |
2019年 | 2篇 |
2018年 | 5篇 |
2017年 | 4篇 |
2016年 | 8篇 |
2015年 | 4篇 |
2014年 | 4篇 |
2013年 | 17篇 |
2012年 | 20篇 |
2011年 | 15篇 |
2010年 | 9篇 |
2009年 | 7篇 |
2008年 | 16篇 |
2007年 | 15篇 |
2006年 | 14篇 |
2005年 | 17篇 |
2004年 | 12篇 |
2003年 | 8篇 |
2002年 | 9篇 |
2001年 | 6篇 |
2000年 | 8篇 |
1999年 | 7篇 |
1998年 | 6篇 |
1997年 | 3篇 |
1996年 | 2篇 |
1995年 | 8篇 |
1994年 | 8篇 |
1993年 | 11篇 |
1992年 | 9篇 |
1991年 | 7篇 |
1990年 | 10篇 |
1989年 | 2篇 |
1988年 | 5篇 |
1987年 | 3篇 |
1986年 | 8篇 |
1985年 | 8篇 |
1984年 | 5篇 |
1983年 | 5篇 |
1982年 | 3篇 |
1981年 | 2篇 |
1980年 | 4篇 |
1979年 | 3篇 |
1976年 | 1篇 |
1975年 | 1篇 |
1970年 | 1篇 |
1959年 | 1篇 |
1947年 | 3篇 |
1930年 | 1篇 |
1927年 | 1篇 |
排序方式: 共有334条查询结果,搜索用时 15 毫秒
21.
Bürli RW McMinn D Kaizerman JA Hu W Ge Y Pack Q Jiang V Gross M Garcia M Tanaka R Moser HE 《Bioorganic & medicinal chemistry letters》2004,14(5):1253-1257
Novel DNA minor-groove binding ligands with a promising antibacterial profile are described. Apart from excellent in vitro potency against multiple Gram-positive bacterial strains such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VRE), and penicillin-intermediate Streptococcus pneumoniae (PISP), a small subset of compounds was active against Gram-negative bacteria such as Escherichia coli (E. coli). 相似文献
22.
Mackiewicz M Nikonova EV Zimmerman JE Galante RJ Zhang L Cater JR Geiger JD Pack AI 《Journal of neurochemistry》2003,85(2):348-357
Adenosine plays a role in promoting sleep, an effect that is thought to be mediated in the basal forebrain. Adenosine levels vary in this region with prolonged wakefulness in a unique way. The basis for this is unknown. We examined, in rats, the activity of the major metabolic enzymes for adenosine - adenosine deaminase, adenosine kinase, ecto- and cytosolic 5'-nucleotidase - in sleep/wake regulatory regions as well as cerebral cortex, and how the activity varies across the day and with sleep deprivation. There were robust spatial differences for the activity of adenosine deaminase, adenosine kinase, and cytosolic and ecto-5'-nucleotidase. However, the basal forebrain was not different from other sleep/wake regulatory regions apart from the tuberomammillary nucleus. All adenosine metabolic enzymes exhibited diurnal variations in their activity, albeit not in all brain regions. Activity of adenosine deaminase increased during the active period in the ventrolateral pre-optic area but decreased significantly in the basal forebrain. Enzymatic activity of adenosine kinase and cytosolic-5'-nucleotidase was higher during the active period in all brain regions tested. However, the activity of ecto-5'-nucleotidase was augmented during the active period only in the cerebral cortex. This diurnal variation may play a role in the regulation of adenosine in relationship to sleep and wakefulness across the day. In contrast, we found no changes specifically with sleep deprivation in the activity of any enzyme in any brain region. Thus, changes in adenosine with sleep deprivation are not a consequence of alterations in adenosine enzyme activity. 相似文献
23.
24.
25.
Lam KS Gustavson DR Pirnik DL Pack E Bulanhagui C Mamber SW Forenza S Stodieck LS Klaus DM 《Journal of industrial microbiology & biotechnology》2002,29(6):299-302
The effect of space flight on production of the antibiotic actinomycin D by Streptomyces plicatus WC56452 was examined onboard the US Space Shuttle mission STS-80. Paired space flight and ground control samples were similarly
prepared using identical hardware, media, and inoculum. The cultures were grown in defined and complex media under dark, anaerobic,
thermally controlled (20°C) conditions with samples fixed after 7 and 12 days in orbit, and viable residuals maintained through
landing at 17 days, 15 h. Postflight analyses indicated that space flight had reduced the colony-forming unit (CFU) per milliliter
count of S. plicatus and increased the specific productivity (pg CFU−1) of actinomycin D. The antibiotic compound itself was not affected, but its production time course was altered in space.
Viable flight samples also maintained their sporulation ability when plated on agar medium postflight, while the residual
ground controls did not sporulate.
Received 21 August 2001/ Accepted in revised form 30 July 2002 相似文献
26.
A Clostridium thermocellum gene, xynX, coding for a xylanase was cloned and the complete nucleotide sequence was determined. The xylanase gene of Clostridium thermocellum consists of an ORF of 3261 nucleotide encoding a xylanase (XynX) of 1087 amino acid residues (116 kDa). Sequence analysis
of XynX showed a multidomain structure that consisted of four different domains: an N-terminal thermostabilizing domain homologous
to sequences found in several thermophilic enzymes, a catalytic domain homologous to family 10 glycosyl hydrolases, a duplicated
cellulose-binding domain (CBD) homologous to family IX CBDs, and a triplicated S-layer homologous domain. A deletion mutant
of xynX having only the catalytic region produced a mutant enzyme XynX-C which retained catalytic activity but lost thermostability.
In terms of half-life at 70 °C, the thermostability of XynX-C was about six times lower than that of the other mutant enzyme,
XynX-TC, produced by a mutant containing both the thermostabilizing domain and the catalytic domain. The optimum temperature
of XynX-C was about 5–10 °C lower than that of XynX-TC.
Received: 12 January 2000 / Received revision: 24 April 2000 / Accepted: 1 May 2000 相似文献
27.
The development of safe and effective gene delivery agents poses a great challenge in the quest to make human gene therapy a reality. Cationic polymers represent one important class of materials for gene delivery, but to date they have shown only moderate efficiency. Improving the efficiency will require the design of new polymers incorporating optimized gene delivery properties. For example, inefficient release of the DNA/polymer complex from endocytic vesicles into the cytoplasm is one of the primary causes of poor gene delivery. Here we report the synthesis of a biocompatible, imidazole-containing polymer designed to overcome this obstacle. DNA/polymer polyplexes incorporating this polymer were shown to have desirable physico-chemical properties for gene delivery and are essentially nontoxic. Using this system, mammalian cells in vitro were transfected in the absence of any exogenous endosomolytic agent such as chloroquine. 相似文献
28.
Hanson RL Howell JM LaPorte TL Donovan MJ Cazzulino DL Zannella V Montana MA Nanduri VB Schwarz SR Eiring RF Durand SC Wasylyk JM Parker WL Liu MS Okuniewicz FJ Chen B Harris JC Natalie KJ Ramig K Swaminathan S Rosso VW Pack SK Lotz BT Bernot PJ Rusowicz A Lust DA Tse KS Venit JJ Szarka LJ Patel RN 《Enzyme and microbial technology》2000,26(5-6):348-358
Allysine ethylene acetal [(S)-2-amino-5-(1,3-dioxolan-2-yl)-pentanoic acid (2)] was prepared from the corresponding keto acid by reductive amination using phenylalanine dehydrogenase (PDH) from Thermoactinomyces intermedius ATCC 33205. Glutamate, alanine, and leucine dehydrogenases, and PDH from Sporosarcina species (listed in order of increasing effectiveness) also gave the desired amino acid but were less effective. The reaction requires ammonia and NADH. NAD produced during the reaction was recyled to NADH by the oxidation of formate to CO(2) using formate dehydrogenase (FDH). PDH was produced by growth of T. intermedius ATCC 33205 or by growth of recombinant Escherichia coli or Pichia pastoris expressing the Thermoactinomyces enzyme. Using heat-dried T. intermedius as a source of PDH and heat-dried Candida boidinii SC13822 as a source of FDH,98%, but production of T. intermedius could not be scaled up. Using heat-dried recombinant E. coli as a source of PDH and heat-dried Candida boidinii 98%. In a third generation process, heat-dried methanol-grown P. pastoris expressing endogenous FDH and recombinant Thermoactinomyces98% ee. 相似文献
29.
Joshua?C.?Snyder Thomas?F.?Pack Lauren?K.?Rochelle Subhasish?K.?Chakraborty Ming?Zhang Andrew?W.?Eaton Yushi?Bai Lauren?A.?Ernst Larry?S.?Barak Alan?S.?Waggoner Marc?G.?CaronEmail author 《BMC biology》2015,13(1):107
Background
Membrane proteins regulate a diversity of physiological processes and are the most successful class of targets in drug discovery. However, the number of targets adequately explored in chemical space and the limited resources available for screening are significant problems shared by drug-discovery centers and small laboratories. Therefore, a low-cost and universally applicable screen for membrane protein trafficking was developed.Results
This high-throughput screen (HTS), termed IRFAP-HTS, utilizes the recently described MarsCy1-fluorogen activating protein and the near-infrared and membrane impermeant fluorogen SCi1. The cell surface expression of MarsCy1 epitope-tagged receptors can be visualized by simple addition of SCi1. User-friendly, rapid, and quantitative detection occurs on a standard infrared western-blotting scanner. The reliability and robustness of IRFAP-HTS was validated by confirming human vasopressin-2 receptor and dopamine receptor-2 trafficking in response to agonist or antagonist. The IRFAP-HTS screen was deployed against the leucine-rich G protein-coupled receptor-5 (Lgr5). Lgr5 is expressed in stem cells, modulates Wnt/ß-catenin signaling, and is therefore a promising drug target. However, small molecule modulators have yet to be reported. The constitutive internalization of Lgr5 appears to be one primary mode through which its function is regulated. Therefore, IRFAP-HTS was utilized to screen 11,258 FDA-approved and drug-like small molecules for those that antagonize Lgr5 internalization. Glucocorticoids were found to potently increase Lgr5 expression at the plasma membrane.Conclusion
The IRFAP-HTS platform provides a versatile solution for screening more targets with fewer resources. Using only a standard western-blotting scanner, we were able to screen 5,000 compounds per hour in a robust and quantitative assay. Multi-purposing standardly available laboratory equipment eliminates the need for idiosyncratic and more expensive high-content imaging systems. The modular and user-friendly IRFAP-HTS is a significant departure from current screening platforms. Small laboratories will have unprecedented access to a robust and reliable screening platform and will no longer be limited by the esoteric nature of assay development, data acquisition, and post-screening analysis. The discovery of glucocorticoids as modulators for Lgr5 trafficking confirms that IRFAP-HTS can accelerate drug-discovery and drug-repurposing for even the most obscure targets.30.
Functional selectivity (or biased agonism) is a property exhibited by some G protein-coupled receptor (GPCR) ligands, which results in the modulation of a subset of a receptor’s signaling capabilities and more precise control over complex biological processes. The dopamine D2 receptor (D2R) exhibits pleiotropic responses to the biogenic amine dopamine (DA) to mediate complex central nervous system functions through activation of G proteins and β-arrestins. D2R is a prominent therapeutic target for psychological and neurological disorders in which DA biology is dysregulated and targeting D2R with functionally selective drugs could provide a means by which pharmacotherapies could be developed. However, factors that determine GPCR functional selectivity in vivo may be multiple with receptors, ligands and transducers contributing to the process. We have recently described a mutagenesis approach to engineer biased D2R mutants in which G protein-dependent ([Gprot]D2R) and β-arrestin-dependent signaling ([βarr]D2R) were successfully separated (Peterson, et al. PNAS, 2015). Here, permutations of these mutants were used to identify critical determinants of the D2R signaling complex that impart signaling bias in response to the natural or synthetic ligands. Critical residues identified in generating [Gprot]D2R and [βarr]D2R conferred control of partial agonism at G protein and/or β-arrestin activity. Another set of mutations that result in G protein bias was identified that demonstrated that full agonists can impart unique activation patterns, and provided further credence to the concept of ligand texture. Finally, the contributions and interplay between different transducers indicated that G proteins are not aberrantly activated, and that receptor kinase and β-arrestin activities are inextricably linked. These data provide a thorough elucidation of the feasibility and malleability of D2R functional selectivity and point to means by which novel in vivo therapies could be modeled. 相似文献