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2.
    
The homotetrameric and biotin‐binding properties of avidin and streptavidin have been exploited for a myriad of biotechnological applications and theoretical studies. Among the few differences between the two proteins is the capacity of avidin to hydrolyze biotinyl p‐nitrophenyl ester (BNP), as opposed to streptavidin, which fully protects the same pseudosubstrate from hydrolysis. Combined mutagenesis and X‐ray analysis have been used to attempt to understand this diametric difference in activities. It was found that a charged residue and one of the loops (L3,4) are together responsible for this difference. Recently, the avidin‐related analogue AVR4 was found to have an even more pronounced BNP‐hydrolysis activity than avidin. Again, the combination of charged residue(s) (Asp39 and/or Arg112) and the rigid conformation of the L3,4 loop was suggested to be responsible for the observed hydrolysis reaction. However, replacement of the latter charged residues in AVR4 resulted in only a modest reduction in hydrolytic activity at most, whereas replacement of the L3,4 loop of avidin with the rigid loop of AVR4 caused a dramatic increase in the activity of avidin. These results clearly demonstrate that the main feature responsible for the observed differences in rates of hydrolysis among the avidins is the conformational status of the L3,4 loop, which imposes conformational constraints on the pseudosubstrate, thereby rendering it susceptible to nucleophilic attack by solvent. In this context, the hydrolytic properties of the avidins reflect enzyme catalysis, in that subtleties in substrate binding are the determining features of catalytic efficiency.  相似文献   
3.
Grazing refuges, external avoidance of herbivory and plant diversity   总被引:16,自引:1,他引:16  
Avoidance and tolerance are the two means by which plants cope with herbivores. Avoidances internal to the plant, such as morphology, chemical repellants, thorns, etc., have received considerable attention in the plant‐herbivore literature, but relatively little consideration has been given to avoidances external to the plant. We develop a conceptual framework of external plant avoidances of herbivory based on foraging selection impedances (associational avoidances), behavioral impedances (indirect avoidances), and physical impedances (refuges) organized along axes of efficiency, degree of protection, and necessity of tolerance characteristics. Associational avoidances are uncommon for terrestrial mammalian herbivores compared to plant‐insect or marine situations. Indirect avoidances mediated through herbivore territoriality, predator avoidance, and other behaviors independent of foraging decisions are probably common in nature, but few have been formally documented. Biotic and geologic refuges providing a physical impedance are the only avoidances shown to have implications for plant biodiversity. This is particularly true for geologic refuges, where there is not a tradeoff between competition and the refuge effect. Small geologic refuges (rock outcrops, cliffs, etc.) are more likely to also positively or negatively alter associated plant microenvironments than large geologic refuges (mesas, islands, etc.). In a survey, 86% of small refuge studies reported positive effects on plant diversity compared to 50% for large refuges. Geologic refuges in more productive environments were more important in protecting diversity than refuges in less productive, semiarid environments, and the effects of protection were greater in communities with short compared to long evolutionary histories of grazing. Other characteristics of refuges such as extent across the landscape and the manner they alter or ameliorate the environment, as well as characteristics of the herbivore such as small or large, generalist or specialist may also determine the effectiveness of refuges, but there are too few studies to assess these factors.  相似文献   
4.

Background

Aortic valve calcification (AVC) secondary to renal failure (RF) is an inflammation-regulated process, but its pathogenesis remains unknown. We sought to assess the cellular processes that are involved in the early phases of aortic valve disease using a unique animal model of RF-associated AVC.

Methods

Aortic valves were obtained from rats that were fed a uremia-inducing diet exclusively for 2, 3, 4, 5, and 6 weeks as well as from controls. Pathological examination of the valves included histological characterization, von Kossa staining, and antigen expression analyses.

Results

After 2 weeks, we noted a significant increase in urea and creatinine levels, reflecting RF. RF parameters exacerbated until the Week 5 and plateaued. Whereas no histological changes or calcification was observed in the valves of any study group, macrophage accumulation became apparent as early as 2 weeks after the diet was started and rose after 3 weeks. By western blot, osteoblast markers were expressed after 2 weeks on the diet and decreased after 6 weeks. Collagen 3 was up-regulated after 3 weeks, plateauing at 4 weeks, whereas collagen 1 levels peaked at 2 and 4 weeks. Fibronectin levels increased gradually until Week 5 and decreased at 6 weeks. We observed early activation of the ERK pathway, whereas other pathways remained unchanged.

Conclusions

We concluded that RF induces dramatic changes at the cellular level, including macrophage accumulation, activation of cell signaling pathway and extracellular matrix modification. These changes precede valve calcification and may increase propensity for calcification, and have to be investigated further.  相似文献   
5.
6.

Background

Mood stabilizers used for treating bipolar disorder (BD) selectively downregulate arachidonic acid (AA) turnover (deacylation–reacylation) in brain phospholipids, when given chronically to rats. In vitro studies suggest that one of these, valproic acid (VPA), which is teratogenic, reduces AA turnover by inhibiting the brain long-chain acyl-CoA synthetase (Acsl)4 mediated acylation of AA to AA-CoA. We tested whether non-teratogenic VPA analogues might also inhibit Acsl4 catalyzed acylation, and thus have a potential anti-BD action.

Methods

Rat Acsl4-flag protein was expressed in Escherichia coli, and the ability of three VPA analogues, propylisopropylacetic acid (PIA), propylisopropylacetamide (PID) and N-methyl-2,2,3,3-tetramethylcyclopropanecarboxamide (MTMCD), and of sodium butyrate, to inhibit conversion of AA to AA-CoA by Acsl4 was quantified using Michaelis–Menten kinetics.

Results

Acsl4-mediated conversion of AA to AA-CoA in vitro was inhibited uncompetitively by PIA, with a Ki of 11.4 mM compared to a published Ki of 25 mM for VPA, while PID, MTMCD and sodium butyrate had no inhibitory effect.

Conclusions

PIA's ability to inhibit conversion of AA to AA-CoA by Acsl4 in vitro suggests that, like VPA, PIA may reduce AA turnover in brain phospholipids in unanesthetized rats, and if so, may be effective as a non-teratogenic mood stabilizer in BD patients.  相似文献   
7.
A laboratory culture of an Israeli benthic midge, Chironomus luridus, was exposed to two chemicals: copper and monochloramine. The objective of this study was to determine the protective nature of Chironomus' larval tube. Three experimental conditions were tested: larva with sand tubes, with silt tubes and without tubes. Larvae without tubes were significantly more sensitive to copper and chloramine than larvae that had sand or silt as tube building substrate. The tubes protected the insects against chemicals throughout 14 days of exposure time. Silt tubes had higher protective value than sand tubes, especially when exposed to copper for a short period of time (LC50/ 24 h, with silt, sand, or none: 80.0, 7.0 and 3.4 mg l–1 copper, respectively). C. luridus seemed to be better protected against copper than against chloramine (LC50/ 24 h, with silt, sand, or none: 12.2, 6.4 and 3.7 mg l–1 chloramine, respectively). The acute toxicity of copper to chironomid larvae was investigated using a cytochemical method. Larva in silt tubes had significantly higher non-specific esterase activity in midgut cells than larvae without tubes. We conclude that, in addition to its role in feeding, respiration and anti-predation shelter, the C. luridus tube protects its inhabitant from toxic substances.  相似文献   
8.
Vibrio cholerae is a waterborne bacterium native to the aquatic environment. There are over 200 known serogroups yet only two cause cholera pandemics in humans. Direct contact of human sewage with drinking water, sea-born currents and marine transportation, represent modes of dissemination of the bacteria and thus the disease. The simultaneous cholera outbreaks that occur sometimes in distant localities within continental landmasses are puzzling. Here we present evidence that flying, non-biting midges (Diptera; Chironomidae), collected in the air, carry viable non-O1 non-O139 serogroups of V. cholerae. The association of V. cholerae with chironomid egg masses, which serve as a V. cholerae reservoir, was further confirmed. In simulated field experiments, we recorded the transfer of environmental V. cholerae by adult midges from the aquatic environment into bacteria-free water-pools. In laboratory experiments, flying adult midges that emerged from V. cholerae (O1 or O139) contaminated water transferred the green fluorescent protein (GFP)-tagged pathogenic bacteria from one laboratory flasks to another. Our findings show that aerial transfer by flying chironomids may play a role in the dissemination of V. cholerae in nature.  相似文献   
9.
Plants of 25 wild Lycopersicon accessions were screened in the greenhouse for resistance to the whitefly-borne tomato yellow leaf curl virus (TYLCV). High levels of resistance were detected in 7 of 9 accessions of L. peruvianum and in all 5 accessions of L. chilense tested. In contrast, plants of 7 accessions of L. hirsutum and 3 of 4 accessions of L. pimpinellifolium were highly susceptible. Plants of accession CIAS 27 (L. pimpinellifolium) showed moderate resistance to TYLCV.  相似文献   
10.

Background and aims

Due to the high spatial and temporal variation in soil CO2 efflux, terrestrial carbon budgets rely on a detailed understanding of the drivers of soil respiration from a diverse range of ecosystems and climate zones. In this study we aim to evaluate the independent influence of vegetation structure and climate on soil CO2 efflux within cerrado ecosystems.

Methods

We examine the seasonal and diel variation of soil CO2 efflux, including its autotrophic and heterotrophic components, within two adjacent and structurally contrasting woody savannas in central Brazil.

Principle results

We found no significant difference in the annual soil CO2 efflux between the two stands (p?=?0.53) despite a clear disparity in both LAI (p?<?0.01) and leaf litterfall (p?<?0.01). The mean annual loss of carbon from the soil was 17.32(±1.48) Mg C?ha?1 of which approximately 63% was accounted for by autotrophic respiration. The relative contribution of autotrophic respiration varied seasonally between 55% in the wet season to 79% of the total soil CO2 efflux in the dry season. Furthermore, seasonal fluctuations of all the soil respiration components were strongly correlated with soil moisture (R 2?=?0.79–0.90, p?<?0.01).

Conclusions

Across these two structurally distinct cerrado stands, seasonal variations in rainfall, was the main driver of soil CO2 efflux and its components. Consequently, soil respiration within these ecosystems is likely to be highly sensitive to any changes in seasonal precipitation patterns.  相似文献   
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