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371.
《Journal of lipid research》2022,63(10):100271
The main fatty acids at the sn-1 position of phospholipids (PLs) are saturated or monounsaturated fatty acids such as palmitic acid (C16:0), stearic acid (C18:0), and oleic acid (C18:1) and are constantly replaced, like unsaturated fatty acids at the sn-2 position. However, little is known about the molecular mechanism underlying the replacement of fatty acids at the sn-1 position, i.e., the sn-1 remodeling. Previously, we established a method to evaluate the incorporation of fatty acids into the sn-1 position of lysophospholipids (lyso-PLs). Here, we used this method to identify the enzymes capable of incorporating fatty acids into the sn-1 position of lyso-PLs (sn-1 lysophospholipid acyltransferase [LPLAT]). Screenings using siRNA knockdown and recombinant proteins for 14 LPLATs identified LPLAT7/lysophosphatidylglycerol acyltransferase 1 (LPGAT1) as a candidate. In vitro, we found LPLAT7 mainly incorporated several fatty acids into the sn-1 position of lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), with weak activities toward other lyso-PLs. Interestingly, however, only C18:0-containing phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were specifically reduced in the LPLAT7-mutant cells and tissues from knockout mice, with a concomitant increase in the level of C16:0- and C18:1-containing PC and PE. Consistent with this, the incorporation of deuterium-labeled C18:0 into PLs dramatically decreased in the mutant cells, while deuterium-labeled C16:0 and C18:1 showed the opposite dynamic. Identifying LPLAT7 as an sn-1 LPLAT facilitates understanding the biological significance of sn-1 fatty acid remodeling of PLs. We also propose to use the new nomenclature, LPLAT7, for LPGAT1 since the newly assigned enzymatic activities are quite different from the LPGAT1s previously reported. 相似文献
372.
Agneta Önfelt 《Chemico-biological interactions》1983,46(2):201-217
Asynchronously growing V79 Chinese hamster cells were treated with colcemid, diamide, carbaryl and methyl mercury, which are all known to be spindle disturbing agents. For each compound the dose response for c-mitosis, survival and level of free sulfhydryl groups was investigated under comparable conditions. Diamide, carbaryl and methyl were all found to give a significant increase of c-mitosis at a dose giving a decrease of non-protein sulfhydryl groups (NPSH, mainly glutathione) of 30–40% suggesting that a decrease of this magnitude may have a predictive value for spindle disturbances. Despite this similarity at concentrations close to the respective thresholds it was found that the c-mitotic activity at higher concentrations was not a simple function of average NPSH decrease. Diamide, which rapidly oxidizes glutathione to glutathione disulfide, was a less efficient c-mitotic agent than carbaryl and methyl mercury in relation to average NPSH decrease at higher concentrations. Protein bound sulfhydryl groups (PSH) were not significantly affected with diamide and carbaryl at their lowest c-mitotic concentrations while methyl mercury caused a significant decrease already at concentrations below the lowest c-mitotic concentration. With colcemid a significant decrease of average NPSH (14%) and PSH (12%) was observed only with concentrations giving close to 100% c-mitotic cells. Concentrations giving more than 20% c-mitosis gave a pronounced decrease of survival with carbaryl, diamide and methyl mercury while no toxic effects were obtained with colcemid, not even with concentrations giving close to 100% c-mitosis. Carbaryl, diamide and methyl mercury caused increased glutathione peroxidase activity indicating that these compounds cause increased lipid peroxidation. The possible connection between peroxidative damage of membranes and c-mitosis is discussed. 相似文献
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375.
Transport and metabolism of 8-14 C-zeatin, applied to an attached de-tipped one-year-old mature leaf of a Yucca plant bearing a bleeding inflorescent stalk, has been studied. Radioactive zeatin ribotide was found in the exudate of the bleeding inflorescence, which was collected over a period of 5 days. Radioactive zeatin ribotide was mainly extracted from the fed leaf. Minor conversion products in this leaf were zeatin ribotide, zeatin- o -β-glucoside and zeatinriboside o -β-glucoside.
In not zeatin fed plants, zeatin- o -β-glucoside was tentatively identified as the main endogenous cytokinin in one-year-old mature leaves. In the bleeding sap of not treated plants no free bases of zeatin or zeatin ribosides were found. After alkaline phosphatase treatment zeatin-riboside was detected by combined gas chromatography-mass spectrometry, indicating the presence of zeatin ribotide in the bleeding sap. High β-glucosidase activity was found in the stern.
Results suggest that stared cytokinin glucosides from Yucca leaves are, converted by β-glucosidase in leaves and stem, transported through the inflorescent stalk as zeatin nucleotides. 相似文献
In not zeatin fed plants, zeatin- o -β-glucoside was tentatively identified as the main endogenous cytokinin in one-year-old mature leaves. In the bleeding sap of not treated plants no free bases of zeatin or zeatin ribosides were found. After alkaline phosphatase treatment zeatin-riboside was detected by combined gas chromatography-mass spectrometry, indicating the presence of zeatin ribotide in the bleeding sap. High β-glucosidase activity was found in the stern.
Results suggest that stared cytokinin glucosides from Yucca leaves are, converted by β-glucosidase in leaves and stem, transported through the inflorescent stalk as zeatin nucleotides. 相似文献
376.
The electrical potential across the cytoplasmic membrane of bacteria can be calculated from the distribution of the lipophilic cation tetraphenylphosphonium between the bulk phases of the medium and the cytoplasm. In order to determine the bulk phase concentrations, information about the binding of the probe to the cellular components is required. In de-energized cells of Rhodopseudomonas sphaeroides the binding appears to be proportional to the free probe concentration. The bulk phase concentrations can only be determined when knowledge is available about the distribution of the binding of the probe over the different cellular components. In this report, models for binding are presented which are based on the assumption that the binding is an energy-independent process. These models allow a proper calculation of the electrical potential when the binding of the probe to the different cellular components is known. 相似文献
377.
Rapid calculation of the solution scattering profile from a macromolecule of known structure 总被引:1,自引:0,他引:1
E E Lattman 《Proteins》1989,5(2):149-155
If one expands the structure factor equation in spherical coordinates, rotational averaging of the molecular Fourier transform, which leads directly to the solution scattering profile, is greatly simplified. It becomes a projection in the polar and azimuthal angular variables. The profile is given by I(R) = 1/2 infinity sigma n = 0 n sigma m = 0 epsilon mNm,n magnitude of Gm,n(R) 2 where Gm,n(R) = sigma jfjYm,n(theta j, phi j)jn(2 pi rjR) The index j runs over all atoms; r, theta, phi are atomic coordinates and epsilon and N are constants; the Ym,n are complex spherical harmonics, and jn are spherical Bessel functions; R = 2 sin theta/lambda. The effects of solvent have been modeled by subtracting from each protein atom a properly weighted water. Hydrogens have been included by using scattering curves fj derived from the spherical averaging of protein atoms with their attached hydrogens. This approach may also be satisfactory for neutron scattering. Published scattering profiles for lysozyme and BPTI have been accurately matched in less than one-tenth the time required by other methods. Separate, adjustable temperature factors for the protein, solvent waters, and bound waters are used, and appear to be needed. In the case of BPTI, as suggested by NMR observations, the observed diffraction pattern was much better accounted for by including only 4 tightly bound waters rather than the roughly 60 seen by crystallography. 相似文献
378.
Emmanuel Van Obberghen Pierre De Meyts Jesse Roth 《Biochimica et Biophysica Acta (BBA)/General Subjects》1979,582(3):486-495
Little is known of the effects of the solvent on hormone-receptor interactions. In the present study the effect of the polar solvent dimethyl sulfoxide on the binding of insulin to its surface receptors on cultured human lymphocytes of the IM-9 line was investigated. At concentrations exceeding 0.1% (v/v), dimethyl sulfoxide produced a dose-related inhibition of 125I-labeled insulin binding. Insulin binding was totally abolished in 20% dimethyl sulfoxide. This inhibition was immediately present and was totally reversible. Analysis of the data of binding at steady state indicated that the decrease in binding of 125I-labeled insulin was due to a reduced affinity of the insulin receptor without noticeable change in the concentration of receptor sites. Kinetic studies showed that the decreased affinity could largely be accounted for by a decreased association rate constant; effects on dissociation and negative cooperativity of the insulin receptor were affected to a much lesser extent. 相似文献
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