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1.
Assessment of uncoupling activity of uncoupling protein 3 using a yeast heterologous expression system. 总被引:2,自引:0,他引:2
Uncoupling protein 3L, uncoupling protein 1 and the mitochondrial oxoglutarate carrier were expressed in Saccharomyces cerevisae. Effects on different parameters related to the energy expenditure were studied. Both uncoupling protein 3L and uncoupling protein 1 reduced the growth rate by 49% and 32% and increased the whole yeast O2 consumption by 31% and 19%, respectively. In isolated mitochondria, uncoupling protein 1 increased the state 4 respiration by 1.8-fold, while uncoupling protein 3L increased the state 4 respiration by 1.2-fold. Interestingly, mutant uncoupling protein 1 carrying the H145Q and H147N mutations, previously shown to markedly decrease the H+ transport activity of uncoupling protein 1 when assessed using a proteoliposome system (Bienengraeber et al. (1998) Biochem. 37, 3-8), uncoupled the mitochondrial respiration to almost the same degree as wild-type uncoupling protein 1. Thus, absence of this histidine pair in uncoupling protein 2 and uncoupling protein 3 does not by itself rule out the possibility that these carriers have an uncoupling function. The oxoglutarate carrier had no effect on any of the studied parameters. In summary, a discordance exists between the magnitude of effects of uncoupling protein 3L and uncoupling protein 1 in whole yeast versus isolated mitochondria, with uncoupling protein 3L having greater effects in whole yeast and a smaller effect on the state 4 respiration in isolated mitochondria. These findings suggest that uncoupling protein 3L, like uncoupling protein 1, has an uncoupling activity. However, the mechanism of action and/or regulation of the activity of uncoupling protein 3L is likely to be different. 相似文献
2.
A comparison of the respiratory chain in particles from Paracoccus denitrificans and bovine heart mitochondria by EPR spectroscopy 总被引:6,自引:0,他引:6
S P Albracht H W van Verseveld W R Hagen M L Kalkman 《Biochimica et biophysica acta》1980,593(2):173-186
A study is presented on the EPR characteristics of the paramagnetic groups in the respiratory chain present in membrane particles of Paracoccus denitrificans, the respiratory system of which is very similar to that in submitochondrial particles from beef heart. All paramagnetic prosthetic groups of the mitochondrial system are also found in the bacterial plasma membrane. Their properties suggest that the respiratory groups are embedded in very similar protein environments in the two systems. 相似文献
3.
Peter-Leon Hagedoorn Laura van der Weel Wilfred R. Hagen 《Journal of visualized experiments : JoVE》2014,(93)
Electron Paramagnetic Resonance (EPR) monitored redox titrations are a powerful method to determine the midpoint potential of cofactors in proteins and to identify and quantify the cofactors in their detectable redox state.The technique is complementary to direct electrochemistry (voltammetry) approaches, as it does not offer information on electron transfer rates, but does establish the identity and redox state of the cofactors in the protein under study. The technique is widely applicable to any protein containing an electron paramagnetic resonance (EPR) detectable cofactor.A typical titration requires 2 ml protein with a cofactor concentration in the range of 1-100 µM. The protein is titrated with a chemical reductant (sodium dithionite) or oxidant (potassium ferricyanide) in order to poise the sample at a certain potential. A platinum wire and a Ag/AgCl reference electrode are connected to a voltmeter to measure the potential of the protein solution. A set of 13 different redox mediators is used to equilibrate between the redox cofactors of the protein and the electrodes. Samples are drawn at different potentials and the Electron Paramagnetic Resonance spectra, characteristic for the different redox cofactors in the protein, are measured. The plot of the signal intensity versus the sample potential is analyzed using the Nernst equation in order to determine the midpoint potential of the cofactor. 相似文献
4.
Reduced hepatic alpha-tocopherol content after long-term administration of ethanol to rats 总被引:1,自引:0,他引:1
G E Bj?rneboe A Bj?rneboe B F Hagen J M?rland C A Drevon 《Biochimica et biophysica acta》1987,918(3):236-241
We have studied the effects of long-term administration of ethanol on the distribution and pharmacokinetics of alpha-tocopherol. In rats fed ethanol (35% of total energy) for 5-6 weeks concentration of alpha-tocopherol in whole liver was reduced by 25% as compared to the pair-fed controls (P less than 0.003). This reduction was significant in the parenchymal cells (28%, P less than 0.004), whereas no significant difference was observed for the nonparenchymal cells. Mitochondrial alpha-tocopherol content was reduced by 55% in the ethanol-treated rats as compared to the controls (P less than 0.002), whereas no significant difference was observed in microsomes, light mitochondria or cytosol. The serum levels of alpha-tocopherol showed no significant difference between the groups. When in vivo labeled chylomicron alpha-[3H]tocopherol was injected intravenously to anesthetized rats, we found a significant increase in serum half-life of alpha-tocopherol in the ethanol-treated group as compared to the controls (P less than 0.025). Hepatic alpha-[3H]tocopherol content was similar in the two groups 24 h after injection. 相似文献
5.
Secretion of alpha-tocopherol from cultured rat hepatocytes 总被引:1,自引:0,他引:1
A Bj?rneboe G E Bj?rneboe B F Hagen J O Nossen C A Drevon 《Biochimica et biophysica acta》1987,922(2):199-205
Primary cultures of rat hepatocytes and rat liver perfusions were used to study hepatic secretion of alpha-tocopherol. The secretion of alpha-tocopherol from hepatocytes in culture was linear with time for 4 h. Ultracentrifugation of the medium revealed that 89.4 +/- 2.1% of alpha-tocopherol secreted during 4 h incubation was associated with the very-low density lipoprotein fraction (VLDL, d less than 1.006 g/ml). Oleic acid had no significant effect on the secretory rate of alpha-tocopherol, whereas eicosapentaenoic acid reduced the amount of alpha-tocopherol secreted to 48.4 +/- 12.7% of the control value after 20 h incubation (P less than 0.01). Monensin, a known inhibitor of VLDL secretion, reduced the secretion of alpha-tocopherol to 14.1 +/- 4.3% of the control value (P less than 0.02). Colchicine and chloroquine inhibited the secretion of alpha-tocopherol in the same order of magnitude as monensin. Hepatic perfusion after intravenous injection of in vivo labeled alpha-[3H]tocopherol lymph, showed that about 75% of the secreted radioactivity was in the VLDL fraction. From these results we conclude that most alpha-tocopherol is secreted from the liver associated with nascent VLDL in rats. 相似文献
6.
Joel B. Hagen 《Biology & philosophy》1989,4(4):433-455
Ecology has often been characterized as an immature scientific discipline. This paper explores some of the sources of this alleged immaturity. I argue that the perception of immaturity results primarily from the fact that historically ecologists have based their work upon two very different approaches to research. 相似文献
7.
8.
The genes coding for human pro alpha 1(IV) collagen and pro alpha 2(IV) collagen are both located at the end of the long arm of chromosome 13. 总被引:5,自引:4,他引:1 下载免费PDF全文
C D Boyd S E Toth-Fejel I K Gadi M Litt M R Condon M Kolbe I K Hagen M Kurkinen J W Mackenzie E Magenis 《American journal of human genetics》1988,42(2):309-314
We have isolated and characterized a cDNA clone containing DNA sequences coding for the noncollagenous carboxy-terminal domain of human pro alpha 2(IV) collagen. Using this cDNA clone in both Southern blot analysis of DNA isolated from human-mouse somatic-cell hybrids and in situ hybridization of normal human metaphase chromosomes, we have demonstrated that the gene coding for human pro alpha 2(IV) collagen is located at 13q33----34, in the same position on chromosome 13 as the pro alpha 1(IV) collagen gene. 相似文献
9.
In an effort to suppress the tuliptree aphidIllinoia liriodendri (Monell), approximately 2,000 eggs ofChrysoperla carnea (Stephens) from a commercial insectary were released 4 times on each of 8 tuliptreesLiriodendron tulipifera L. in Berkeley, California, during the spring of 1984.
On trees foraged by the Argentine antIridomyrmex humilis (Mayr), 98% of the eggs ofC. carnea were removed from the egg release tapes by the ants. A total of about 1,250 larvae per tree eclosed from the 8,000 eggs released
on each tree without ants. Fifty percent of the larvae that did eclose died due to cannibalism or entrapment in the sticky
egg release tapes and approximately 625 first instar larvae on each tree were free to forage for aphids.
Inundative lacewing releases ofC. carnea did not suppress populations ofI. liriodendri due to ant predation, the low viability of commercial eggs (0–73% eclosion),
Résumé Dans le but de limiter les populations du puceron du tulipierIllinoia liriodendri (Monell), 4 lachers d'environ 2.000 œufs de provenance commerciale deChrysoperla carnea (Stephens) ont été réalisés au cours du printemps 1984 sur 8 tulipiersLiriodendron tulipifera L., à Berkeley en Californie. cannibalism by emerged larvae, and inadequate release technology. Sur les arbres visités par la fourmi d'ArgentineIridomyrmex humilis (Mayr), 98% des œufs deC. carnea ont été enlevés du support artificiel par les fourmis. Sur les arbres exempts de fourmis, la mortalité de 50% des larves est due au cannibalisme ou à leur engluement sur le support de lacher. A partir de 8.000 œufs déposés sur chaque arbre sans fourmi, on aboutit à environ 625 larves de 1er stade susceptibles de rechercher des pucerons. Les lachers inondatifs deC. carnea n'ont pas limité les populations d'I. liriodendri. Les raisons en sont: la consommation par les fourmis, une faible viabilité de la plus grande partie des œufs commercialisés (0–73% d'éclosions), une technique inadaptée pour le lacher des œufs et le cannibalisme par les larves elles-mêmes deC. carnea.相似文献
10.
J M Aiken F D Miller F Hagen D I McKenzie S A Krawetz J H van de Sande J B Rattner G H Dixon 《Biochemistry》1985,24(22):6268-6276
We have located an extensive (AC)n-rich but specific sequence downstream of three rainbow trout protamine genes. Although sharing considerable sequence homology, including a perfectly conserved 46 base pair repeat, the sequences exhibit a regular heterogeneity in the length of the (AC)n-rich tracts. Radioimmunoassay experiments, S1 nuclease sensitivity studies, two-dimensional electrophoretic analysis, and immunoelectron microscopy studies have been used to determine if the region could assume a Z DNA conformation. It was found that, in a supercoiled plasmid, the (AC)n-rich region has the ability to attain the Z DNA conformation under physiological conditions. 相似文献