全文获取类型
收费全文 | 746篇 |
免费 | 84篇 |
国内免费 | 2篇 |
专业分类
832篇 |
出版年
2021年 | 18篇 |
2020年 | 7篇 |
2019年 | 9篇 |
2018年 | 8篇 |
2017年 | 14篇 |
2016年 | 31篇 |
2015年 | 60篇 |
2014年 | 60篇 |
2013年 | 88篇 |
2012年 | 24篇 |
2011年 | 18篇 |
2010年 | 52篇 |
2009年 | 30篇 |
2008年 | 20篇 |
2007年 | 6篇 |
2006年 | 18篇 |
2005年 | 10篇 |
2004年 | 8篇 |
2003年 | 19篇 |
2002年 | 9篇 |
2001年 | 6篇 |
2000年 | 14篇 |
1999年 | 13篇 |
1998年 | 17篇 |
1997年 | 9篇 |
1996年 | 18篇 |
1995年 | 16篇 |
1994年 | 9篇 |
1993年 | 11篇 |
1992年 | 8篇 |
1991年 | 10篇 |
1990年 | 5篇 |
1989年 | 6篇 |
1988年 | 5篇 |
1987年 | 10篇 |
1986年 | 5篇 |
1985年 | 10篇 |
1984年 | 7篇 |
1981年 | 9篇 |
1978年 | 6篇 |
1977年 | 7篇 |
1974年 | 5篇 |
1973年 | 10篇 |
1972年 | 9篇 |
1971年 | 7篇 |
1970年 | 8篇 |
1969年 | 5篇 |
1966年 | 5篇 |
1965年 | 10篇 |
1963年 | 5篇 |
排序方式: 共有832条查询结果,搜索用时 15 毫秒
51.
Background
Because of the difficulties involved in learning and using 3D modeling and rendering software, many scientists hire programmers or animators to create models and animations. This both slows the discovery process and provides opportunities for miscommunication. Working with multiple collaborators, a tool was developed (based on a set of design goals) to enable them to directly construct models and animations.Results
SketchBio is presented, a tool that incorporates state-of-the-art bimanual interaction and drop shadows to enable rapid construction of molecular structures and animations. It includes three novel features: crystal-by-example, pose-mode physics, and spring-based layout that accelerate operations common in the formation of molecular models. Design decisions and their consequences are presented, including cases where iterative design was required to produce effective approaches.Conclusions
The design decisions, novel features, and inclusion of state-of-the-art techniques enabled SketchBio to meet all of its design goals. These features and decisions can be incorporated into existing and new tools to improve their effectiveness.Electronic supplementary material
The online version of this article (doi:10.1186/1471-2105-15-334) contains supplementary material, which is available to authorized users. 相似文献52.
Richard A. Hurt Jr Michael S. Robeson II Migun Shakya James G. Moberly Tatiana A. Vishnivetskaya Baohua Gu Dwayne A. Elias 《PloS one》2014,9(7)
Despite over three decades of progress, extraction of high molecular weight (HMW) DNA from high clay soils or iron oxide cemented clay has remained challenging. HMW DNA is desirable for next generation sequencing as it yields the most comprehensive coverage. Several DNA extraction procedures were compared from samples that exhibit strong nucleic acid adsorption. pH manipulation or use of alternative ion solutions offered no improvement in nucleic acid recovery. Lysis by liquid N2 grinding in concentrated guanidine followed by concentrated sodium phosphate extraction supported HMW DNA recovery from clays high in iron oxides. DNA recovered using 1 M sodium phosphate buffer (PB) as a competitive desorptive wash was 15.22±2.33 µg DNA/g clay, with most DNA consisting of >20 Kb fragments, compared to 2.46±0.25 µg DNA/g clay with the Powerlyzer system (MoBio). Increasing PB concentration in the lysis reagent coincided with increasing DNA fragment length during initial extraction. Rarefaction plots of 16S rRNA (V1–V3 region) pyrosequencing from A-horizon and clay soils showed an ∼80% and ∼400% larger accessed diversity compared to the Powerlyzer soil DNA system, respectively. The observed diversity from the Firmicutes showed the strongest increase with >3-fold more operational taxonomic units (OTU) recovered. 相似文献
53.
Of 43 older patients (aged 65 to 83 years) with arteriosclerosis obliterans treated by lumbar sympathectomy for one or both limbs, 19 had excellent result, 13 had fair result and four had poor result. One died postoperatively and six later. Results were better than prognosticated from response to sympathetic block. Thirty-four patients considered the operation worth while and twelve, after unilateral sympathectomy, requested operation for the other limb also. Twenty-four after operation could walk farther without claudication. 相似文献
54.
Bennett T. Farmer II Luciano Müller Edward P. Nikonowicz Arthur Pardi 《Journal of biomolecular NMR》1994,4(1):129-133
Summary A set of three 3D (1H, 13C, 15N) triple-resonance correlation experiments has been designed to provide H1-H8 intraresidue sugar-to-base correlations in purines in an unambiguous and efficient manner. Together, the HsCsNb, HsCs(N)bCb, and HbNbCb experiments correlate the H1 sugar proton to the H8 proton of the attached base by means of the {H1, C1, N9, C8, H8} heteronuclear scalar coupling network. The assignment strategy presented here allows for unambiguous H1-H8 intraresidue correlations, provided that no two purines have both the same H1 and C1 chemical shifts and the same C8 and N9 chemical shifts. These experiments have yielded H1-H8 intraresidue sugar-to-base correlations for all five guanosines in the [13C, 15N] isotopically labeled RNA duplex r(GGCGCUUGCGUC)2. 相似文献
55.
Aggregation of amphipathic peptides at an aqueous–organic interface using coarse-grained simulations
The effect of an aqueous/organic interface on the folding and aggregation of amphipathic peptides is examined by applying discontinuous molecular dynamics (DMD) simulations combined with an intermediate resolution protein model, PRIME20, to a peptide/interface system. The systems contain 48 (KLLK)4 peptides in random coil or α-helical conformations interacting with both strong and weak interfaces. In the absence of an interface, most of the oligomers form helical bundles, a small fraction of which convert to β-sheets when the temperature is above the folding transition. Adding a weak interface decreases oligomer formation above the folding temperature and increases it below. Little monolayer formation is observed at the weak interface; instead reversible adsorption increases the local peptide concentration near the interface, promoting helical bundle formation in the aqueous phase below the folding temperature and β-sheet formation above the folding temperature. Introducing a strong interface leads to irreversible adsorption, promoting formation of helical monolayers below the folding temperature and mixed β-sheet/amorphous monolayers above the folding temperature. The (KLLK)4 peptide is more likely to adsorb to the interface when it is in an α-helical conformation, as opposed to a random coil, because of its larger hydrophobic moment. 相似文献
56.
Hoover SW Marner WD Brownson AK Lennen RM Wittkopp TM Yoshitani J Zulkifly S Graham LE Chaston SD McMahon KD Pfleger BF 《Applied microbiology and biotechnology》2011,91(2):435-446
The predominant strategy for using algae to produce biofuels relies on the overproduction of lipids in microalgae with subsequent conversion to biodiesel (methyl-esters) or green diesel (alkanes). Conditions that both optimize algal growth and lipid accumulation rarely overlap, and differences in growth rates can lead to wild species outcompeting the desired lipid-rich strains. Here, we demonstrate an alternative strategy in which cellulose contained in the cell walls of multicellular algae is used as a feedstock for cultivating biofuel-producing microorganisms. Cellulose was extracted from an environmental sample of Cladophora glomerata-dominated periphyton that was collected from Lake Mendota, WI, USA. The resulting cellulose cake was hydrolyzed by commercial enzymes to release fermentable glucose. The hydrolysis mixture was used to formulate an undefined medium that was able to support the growth, without supplementation, of a free fatty acid (FFA)-overproducing strain of Escherichia coli (Lennen et. al 2010). To maximize free fatty acid production from glucose, an isopropyl β-D-1-thiogalactopyranoside (IPTG)-inducible vector was constructed to express the Umbellularia californica acyl-acyl carrier protein (ACP) thioesterase. Thioesterase expression was optimized by inducing cultures with 50 μM IPTG. Cell density and FFA titers from cultures grown on algae-based media reached 50% of those (~90 μg/mL FFA) cultures grown on rich Luria-Bertani broth supplemented with 0.2% glucose. In comparison, cultures grown in two media based on AFEX-pretreated corn stover generated tenfold less FFA than cultures grown in algae-based media. This study demonstrates that macroalgal cellulose is a potential carbon source for the production of biofuels or other microbially synthesized compounds. 相似文献
57.
Comparison of Two Liquid Blood Culture Media Containing Sodium Polyanetholesulfonate: Tryptic Soy and Columbia 下载免费PDF全文
In a comparison of tryptic soy broth and Columbia broth, two blood culture media containing sodium polyanetholesulfonate, there were 589 positive cultures (excluding presumed contaminants). The two media were equivalent in performance except for lower detection rates for Staphylococcus aureus (P < 0.01) and Pseudomonas aeruginosa (P = 0.05) and a higher detection rate for Bacillus (P < 0.01) in Columbia broth. No significant differences were noted in time intervals to detection of positivity. Routine subcultures on the 1st and 5th days of incubation provided the initial detection of 18.1% of the positive cultures. 相似文献
58.
Michelle W. Clark Anna M. Yie Elizabeth K. Eder Richard G. Dennis Preston J. Basting Keith A. Martinez II Brian D. Jones Joan L. Slonczewski 《PloS one》2015,10(12)
Under certain kinds of cytoplasmic stress, Escherichia coli selectively reproduce by distributing the newer cytoplasmic components to new-pole cells while sequestering older, damaged components in cells inheriting the old pole. This phenomenon is termed polar aging or cell division asymmetry. It is unknown whether cell division asymmetry can arise from a periplasmic stress, such as the stress of extracellular acid, which is mediated by the periplasm. We tested the effect of periplasmic acid stress on growth and division of adherent single cells. We tracked individual cell lineages over five or more generations, using fluorescence microscopy with ratiometric pHluorin to measure cytoplasmic pH. Adherent colonies were perfused continually with LBK medium buffered at pH 6.00 or at pH 7.50; the external pH determines periplasmic pH. In each experiment, cell lineages were mapped to correlate division time, pole age and cell generation number. In colonies perfused at pH 6.0, the cells inheriting the oldest pole divided significantly more slowly than the cells inheriting the newest pole. In colonies perfused at pH 7.50 (near or above cytoplasmic pH), no significant cell division asymmetry was observed. Under both conditions (periplasmic pH 6.0 or pH 7.5) the cells maintained cytoplasmic pH values at 7.2–7.3. No evidence of cytoplasmic protein aggregation was seen. Thus, periplasmic acid stress leads to cell division asymmetry with minimal cytoplasmic stress. 相似文献
59.
Data from epidemiological studies might be seen as superior to data from animal bioassays for risk assessment purposes. Because humans are the population of interest, use of epidemiological data avoids interspecies extrapolation. However, one must not assume that an epidemiological study is necessarily valid at face value. We describe issues of validity that arise in the conduct and interpretation of epidemiological research and that affect the utility of epidemiological data in risk assessment. These issues include choice of study design, size and representativeness of the study sample, measurement of exposures and outcomes, control of confounding and specification of statistical model for analysis of data, all of which affect the accuracy and validity of study results. 相似文献
60.
William H. Frey II Susan E. Senogles Leonard L. Heston Vicente B. Tuason Susan E. Nicol 《Journal of neurochemistry》1980,35(6):1418-1430
Abstract: Partial purification of soluble guanylate cyclase on DEAE-Sephacel yields two separate peaks of guanylate cyclase activity. After 10-fold purification of the soluble enzyme, guanylate cyclase is markedly inhibited by micromolar concentrations of dopamine (I50= 0.2 μm). Dopamine inhibition is observed whether the reaction is conducted with Mn21 or with Mg2+, under atmosphere or N2(g), and using enzyme from either peak from the DEAESephacel column. Other catecholamines also inhibit partially purified guanylate cyclase with an order of potency at 1 μm of: dopamine =l -DOPA > norepinephrine = isoproterenol = adrenochrome > epinephrine. The structural requirements for inhibition are two free hydroxyl groups on the phenyl ring and an ethylamine side chain. Dopamine also inhibits the Triton X-100-solubilized microsomal guanylate cyclase after partial purification on DEAESephacel. Neither chlorpromazine, propranolol, nor phentolamine at 20 μm effectively block the dopamine inhibition of partially purified soluble guanylate cyclase. Micromolar concentrations of the reducing agents dithiothreitol and glutathione also inhibit partially purified guanylate cyclase, but unlike these agents, catecholamines can inhibit whether added in the reduced or the oxidized forms. Inhibition of enzyme activity by micromolar concentrations of dopamine, adrenochrome, or dithiothreitol is rapidly reversed by dilution and the dopamine inhibition is competitive with MgGTP. Inhibition does not appear to involve covalent binding or to result from the ability of catecholamines to reduce the concentrations of oxygen or free radicals in solution. 相似文献