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101.
102.
Plant mitochondria contain an alternative oxidase (AOX) acting as a terminal electron acceptor of the alternative pathway
in the electron transport chain. Here we describe the production of inducible antisense Aox1a plants of Arabidopsis thaliana (L.) Heynh. and the procedures used to determine the resulting alternative pathway activity. The Arabidopsis Aox1a cDNA sequence was cloned behind a copper-inducible promoter system in the antisense orientation. Arabidopsis thaliana (Columbia) plants were transformed by in-planta vacuum infiltration with Agrobacterium containing the antisense construct. Whole-leaf ethanol production was used as a measure to investigate alternative pathway
activity in the presence of antimycin A. After 24 h, leaves from the copper-induced, antisense line F1.1 produced up to 8.8
times more ethanol (via aerobic fermentation) than the non-induced and wild-type leaves, indicating effective cytochrome pathway
inhibition by antimycin A and a decreased alternative pathway activity in induced F1.1 leaves. Transgene expression studies
also revealed no expression in non-induced leaves and up until 24 h post-induction. Copper-induced transgenic leaves were
less susceptible to alternative pathway inhibition than non-induced transgenic leaves, as seen via tissue-slice respiratory
studies, and mitochondrial respiration, using F1.1 cell cultures, also supported this. These results demonstrate the successful
production of a transgenic line of Arabidopsis in which the alternative pathway activity can be genetically manipulated with an inducible antisense system.
Received: 31 March 2000 / Accepted: 10 May 2000 相似文献
103.
David JT. Campbell Paul E. Ronksley Braden J. Manns Marcello Tonelli Claudia Sanmartin Robert G. Weaver Deirdre Hennessy Kathryn King-Shier Tavis Campbell Brenda R. Hemmelgarn for the Interdisciplinary Chronic Disease Collaboration 《PloS one》2014,9(4)
Background
Management of chronic diseases requires patients to adhere to recommended health behavior change and complete tests for monitoring. While studies have shown an association between low income and lack of adherence, the reasons why people with low income may be less likely to adhere are unclear. We sought to determine the association between household income and receipt of health behavior change advice, adherence to advice, receipt of recommended monitoring tests, and self-reported reasons for non-adherence/non-receipt.Methods
We conducted a population-weighted survey, with 1849 respondents with cardiovascular-related chronic diseases (heart disease, hypertension, diabetes, stroke) from Western Canada (n = 1849). We used log-binomial regression to examine the association between household income and the outcome variables of interest: receipt of advice for and adherence to health behavior change (sodium reduction, dietary improvement, increased physical activity, smoking cessation, weight loss), reasons for non-adherence, receipt of recommended monitoring tests (cholesterol, blood glucose, blood pressure), and reasons for non-receipt of tests.Results
Behavior change advice was received equally by both low and high income respondents. Low income respondents were more likely than those with high income to not adhere to recommendations regarding smoking cessation (adjusted prevalence rate ratio (PRR): 1.55, 95%CI: 1.09–2.20), and more likely to not receive measurements of blood cholesterol (PRR: 1.72, 95%CI 1.24–2.40) or glucose (PRR: 1.80, 95%CI: 1.26–2.58). Those with low income were less likely to state that non-adherence/non-receipt was due to personal choice, and more likely to state that it was due to an extrinsic factor, such as cost or lack of accessibility.Conclusions
There are important income-related differences in the patterns of health behavior change and disease monitoring, as well as reasons for non-adherence or non-receipt. Among those with low income, adherence to health behavior change and monitoring may be improved by addressing modifiable barriers such as cost and access. 相似文献104.
Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania
MJAM Oesterholt JT Bousema OK Mwerinde C Harris P Lushino A Masokoto H Mwerinde FW Mosha CJ Drakeley 《Malaria journal》2006,5(1):1-7
Background
Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections.Methods
Urine and saliva were obtained from 47 thick film positive and 4 negative individuals one day after collection of blood slides and filter paper blood spots. P. falciparum DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy® kit (urine and saliva only). Blood, urine and saliva extracts were subjected to PCR in separate batches. Amplicons from the various sample types were examined for MSP2 polymorphisms and restriction fragment patterns on DHFR amino acid codon 59.Results and discussion
Malaria infections exhibited primarily low-grade parasite densities, with a geometric mean of 775 asexual parasites/μl. Regularly matching polymorphic MSP2 genotypes were found between the corresponding urine, saliva and peripheral blood amplicons of each individual, with different inter-individual polymorphic genotypes. Amplicon yields were significantly dependent on DNA extraction method, parasite density and primer set (p < 0.001). A Qiagen® kit extraction had more than 2× higher amplicon yield than the Chelex method, for both urine and saliva. Amplicon yields were 1.6 fold higher from saliva than urine. For each unit increase in log parasite density, the probability of amplicon enhanced 1.8 fold. Highest amplicon yields were obtained from the primer set with the shortest PCR product.Conclusion
P. falciparum infection is detectable by PCR on human urine and saliva samples. Subject to further refinement of extraction technique and amplicon yields, large-scale malaria parasite screening and epidemiological surveys could be possible without the need to collect blood and use of needles or sharps. 相似文献105.
James H. Bryce Joaquim Azcon-Bieto Joseph T. Wiskich David A. Day 《Physiologia plantarum》1990,78(1):105-111
The aim was to test the hypothesis that rotenone-insensive electron transport (bypass of complex I) may underlie rapid state 4 (ADP-limited) mitochondrial respiration. A comparison of mitochondria from soybean ( Glycine max L. cv. Bragg) cotyledons and nodules showed that ADP-sufficient (state 3) malate plus pyruvate oxidation by mitochondria from 7-day-old cotyledons was inhibited 50% by rotenone and state 4 rates were rapid, whereas nodule mitochondria were 80% inhibited by rotenone and had slower state 4 rates of malate plus pyruvate oxidation. Respiration of malate alone (pH 7.6) by cotyledon mitochondria was slow, especially in the absence of ADP; subsequent addition of pyruvate dramatically increased state 4 oxygen uptake concomitant with a rapid rise in mitochondrial NADH (determined by fluorimetry). Rotenone had no effect on this increased rate of state 4 respiration. The rate of malate oxidation by nodule mitochondria was relatively rapid compared with cotyledon mitochondria. The addition of pyruvate in state 4 caused a slow increase in matrix NADH and only a slight stimulation of oxygen uptake. Rotenone inhibited state 4 malate plus pyruvate oxidation by 50% in these mitochondria. From a large number of cotyledon and nodule mitochondrial preparations, a close correlation was found between the rate of state 4 oxygen uptake and rotenone-resistance. During cotyledon development increased rotenone-resistance was associated with an increase in the alternative oxidase. Addition of pyruvate to cotyledon mitochondria, during state 4 oxidation of malate in the presence of antimycin A, significantly stimulated O2 uptake and also almost eliminated respiratory control. Such combined operation of the rotenone-insensitive bypass and the alternative oxidase in vivo will significantly affect the extent to which adenylates control the rate of electron transport. 相似文献
106.
2-Oxoglutarate dehydrogenase and pyruvate dehydrogenase activities in plant mitochondria: interaction via a common coenzyme a pool 总被引:3,自引:0,他引:3
2-Oxoglutarate (2-OG)-dependent O2 uptake by washed or purified turnip (Brassica rapa L.) and pea (Pisum sativum L. cv. Massey Gem) leaf mitochondria, in the presence of malonate, was inhibited between 65 and 90% by micromolar levels of pyruvate. The inhibition was not observed in the absence of malonate and was reversed by alpha-cyano-4-hydroxycinnamic acid. The inhibition was also reversed by oxaloacetate or by malate, but not by any other tricarboxylic acid cycle intermediates. The stimulation of O2 uptake by oxaloacetate was half maximal at 8-9 microM and was transient, indicating its action was not mediated through the complete metabolic removal of pyruvate. Pyruvate had not effect on 2-OG oxidation under conditions in which pyruvate dehydrogenase was not active, indicating that pyruvate metabolism, rather than pyruvate itself, was responsible for producing the inhibition of 2-OG oxidation. Similar results were obtained with detergent-treated mitochondrial extracts with the exception that the inhibition of 2-OG oxidation by pyruvate could also be reversed by coenzyme A. The results suggest that pyruvate inhibits 2-oxoglutarate oxidation, in intact plant mitochondria, by sequestering intramitochondrial CoA as acetyl-CoA and, in the absence of citrate synthase activity, reduces the amount of free coenzyme A available for 2-oxoglutarate dehydrogenase. These results indicate that pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase share a common CoA pool within plant mitochondria and that the turnover of the acyl-CoA product of one enzyme will dramatically influence the activity of the other. 相似文献
107.
108.
Characteristics of primary phases in chlorophyll a fluorescence transients based on room temperature in vivo measurement with a Plant Productivity Fluorometer (Brancker Model SF-10) can be greatly facilitated by coupling the instrument to a fast data acquisition system. The SF-10 was linked to a Multitech Industrial Corporation Microprofessor Microcomputer and further modified to ensure simultaneous onset of light activation and signal capture. Circuit diagrams and program listings are given in detail. This microprocessor system is capable of capturing signal changes over a minimum period of 200 milliseconds to a maximum of 6 seconds. Accuracy of recorded data is dependent on rate of change of the input signal and the recording time period. Acquisition and storage of 5000 points from zero to 300 milliseconds ensured clear resolution of Fo, I and D when played back over 120 seconds on a chart recorder. For routine use, the primary transient can be captured over 0–2 seconds and then replayed as an accompaniment to standard slower presentation of primary plus secondary transients. Coincidence of signal amplitude for Fp on both systems can then be ascertained while retaining adequate resolution of Fo and I. 相似文献
109.
Inokuchi Ritsuko; Itagaki Tadashi; Wiskich Joseph T.; Nakayama Katsumi; Okada Mitsumasa 《Plant & cell physiology》1997,38(3):327-335
NADP-glutamate dehydrogenase (EC 1.4.1.4
[EC]
; NADP-GDH) was purifiedto electrophoretic homogeneity from the multinuclear-unicellulargreen marine alga in Sipho-nales, Bryopsis maxima, and its propertieswere examined. Mr of the undenatured enzyme was 280 kDa, andthe enzyme is thought to be a hexamer of 46 kDa subunit protein.Optimum pHs for the reductive amination and oxidative deaminationwere 7.5 and 8.2-9.0 respectively. The enzyme displayed NADPH/NADH-specificactivities with a ratio of 18 :1. Apparent Km values for 2-oxoglutarate,ammonia, NADPH, glutamate and NADP+ were 3.0, 2.2, 0.03, 3.2and 0.01 mM respectively. The enzymochemical characteristicsof the GDH were studied and compared to those of other species.The B. maxima GDH was insensitive to 5 mM Ca2+ and to 1 mM EDTAin contrast to higher plant NAD-GDHs. Chemical modificationswith DTNB and pCMBS suggested that cysteine residues are essentialfor the enzymatic activity as in other species GDHs. The GDHwas not affected by 1 mM purine nucleotides, suggesting thatthe enzyme is not allosteric, in contrast to animal NAD(P)-GDHsand fungal NAD-GDHs. (Received August 12, 1996; Accepted January 7, 1997) 相似文献
110.
Characteristics of primary phases in chlorophyll a fluorescence transients based on room temperature in vivo measurement with a Plant Productivity Fluorometer (Brancker Model SF-10) can be greatly facilitated by coupling the instrument to a fast data acquisition system. The SF-10 was linked to a Multitech Industrial Corporation Microprofessor Microcomputer and further modified to ensure simultaneous onset of light activation and signal capture. Circuit diagrams and program listings are given in detail. This microprocessor system is capable of capturing signal changes over a minimum period of 200 milliseconds to a maximum of 6 seconds. Accuracy of recorded data is dependent on rate of change of the input signal and the recording time period. Acquisition and storage of 5000 points from zero to 300 milliseconds ensured clear resolution of Fo, I and D when played back over 120 seconds on a chart recorder. For routine use, the primary transient can be captured over 0–2 seconds and then replayed as an accompaniment to standard slower presentation of primary plus secondary transients. Coincidence of signal amplitude for Fp on both systems can then be ascertained while retaining adequate resolution of Fo and I. 相似文献