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41.
CPT1c is localized in endoplasmic reticulum of neurons and has carnitine palmitoyltransferase activity 总被引:2,自引:0,他引:2
Sierra AY Gratacós E Carrasco P Clotet J Ureña J Serra D Asins G Hegardt FG Casals N 《The Journal of biological chemistry》2008,283(11):6878-6885
CPT1c is a carnitine palmitoyltransferase 1 (CPT1) isoform that is expressed only in the brain. The enzyme has recently been localized in neuron mitochondria. Although it has high sequence identity with the other two CPT1 isoenzymes (a and b), no CPT activity has been detected to date. Our results indicate that CPT1c is expressed in neurons but not in astrocytes of mouse brain sections. Overexpression of CPT1c fused to the green fluorescent protein in cultured cells demonstrates that CPT1c is localized in the endoplasmic reticulum rather than mitochondria and that the N-terminal region of CPT1c is responsible for endoplasmic reticulum protein localization. Western blot experiments with cell fractions from adult mouse brain corroborate these results. In addition, overexpression studies demonstrate that CPT1c does not participate in mitochondrial fatty acid oxidation, as would be expected from its subcellular localization. To identify the substrate of CPT1c enzyme, rat cDNA was overexpressed in neuronal PC-12 cells, and the levels of acylcarnitines were measured by high-performance liquid chromatography-mass spectrometry. Palmitoylcarnitine was the only acylcarnitine to increase in transfected cells, which indicates that palmitoyl-CoA is the enzyme substrate and that CPT1c has CPT1 activity. Microsomal fractions of PC-12 and HEK293T cells overexpressing CPT1c protein showed a significant increase in CPT1 activity of 0.57 and 0.13 nmol.mg(-1).min(-1), respectively, which is approximately 50% higher than endogenous CPT1 activity. Kinetic studies demonstrate that CPT1c has similar affinity to CPT1a for both substrates but 20-300 times lower catalytic efficiency. 相似文献
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Joseph Bonaventura Celia Bonaventura Gino Amiconi Eraldo Antonini Maurizio Brunori 《Archives of biochemistry and biophysics》1974,161(1):328-332
Hemoglobin Leiden is an abnormal human hemoglobin in which a glutamic acid residue has been deleted from the β-chain at position 6 or 7. The α-amino groups of the β-chain N-termini in tetrameric hemoglobin A are thought to be directly involved in the binding of simple anions and organic phosphates (1). The deletion of the 4th or 5th residue of the A helix in hemoglobin Leiden shortens the N-terminus of the β-chain, and the results reported here show that the anion binding site has been affected. Hemoglobin Leiden shows a decreased response to inorganic phosphate, chloride, 2,3-diphosphoglycerate, and inositol hexaphosphate, both in equilibria and kinetics of ligand binding. Although hemoglobin Leiden shows an altered response to anions, neither the cooperativity of ligand binding nor the Bohr effect are significantly altered by the deletion. The decreased effect of cofactors seems to be due to a decrease in the strength of anion binding which may be attributed to the altered geometry of the anion binding site. 相似文献
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E Chiancone N M Anderson E Antonini J Bonaventura C Bonaventura M Brunori C Spagnuolo 《The Journal of biological chemistry》1974,249(18):5689-5694
47.
Maurizio Brunori Joseph Bonaventura Celia Bonaventura Bruno Giardina Francesco Bossa Eraldo Antonini 《Molecular and cellular biochemistry》1973,1(2):189-196
Summary The diversity of the structural and functional properties of the various components of trout blood may be taken as a type case of molecular adaptation to physiological requirements. Studies on this system yield, in addition, information which appears relevant to the interpretation of the behavior of mammalian hemoglobins.an invited article 相似文献
48.
C Bonaventura G Ferruzzi S Tesh R D Stevens 《The Journal of biological chemistry》1999,274(35):24742-24748
S-Nitrosated hemoglobin (SNO-Hb) is of interest because of the allosteric control of NO delivery from SNO-Hb made possible by the conformational differences between the R- and T-states of Hb. To better understand SNO-Hb, the oxygen binding properties of S-nitrosated forms of normal and sickle cell Hb were investigated. Spectral assays and electrospray ionization mass spectrometry were used to quantify the degree of S-nitrosation. Hb A(0) and unpolymerized Hb S exhibit similar shifts toward their R-state conformations in response to S-nitrosation, with increased oxygen affinity and decreased cooperativity. Responses to 2, 3-diphosphoglycerate were unaltered, indicating regional changes in the deoxy structure of SNO-Hb that accommodate NO adduction. A cycle of deoxygenation/reoxygenation does not cause loss of NO or appreciable heme oxidation. There is, however, appreciable loss of NO and heme oxidation when oxygen-binding experiments are carried out in the presence of glutathione. These results indicate that the in vivo stability of SNO-Hb and its associated vasoactivity depend on the abundance of thiols and other factors that influence transnitrosation reactions. The increased oxygen affinity and R-state character that result from S-nitrosation of Hb S would be expected to decrease its polymerization and thereby lessen the associated symptoms of sickle cell disease. 相似文献
49.
Shift of Clinical Human Immunodeficiency Virus Type 1 Isolates from X4 to R5 and Prevention of Emergence of the Syncytium-Inducing Phenotype by Blockade of CXCR4 总被引:5,自引:0,他引:5
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Jos A. Est Cecilia Cabrera Juli Blanco Arantxa Gutierrez Gary Bridger Geoffrey Henson Bonaventura Clotet Dominique Schols Erik De Clercq 《Journal of virology》1999,73(7):5577-5585
The emergence of X4 human immunodeficiency virus type 1 (HIV-1) strains in HIV-1-infected individuals has been associated with CD4(+) T-cell depletion, HIV-mediated CD8(+) cell apoptosis, and an impaired humoral response. The bicyclam AMD3100, a selective antagonist of CXCR4, selected for the outgrowth of R5 virus after cultivation of mixtures of the laboratory-adapted R5 (BaL) and X4 (NL4-3) HIV strains in the presence of the compound. The addition of AMD3100 to peripheral blood mononuclear cells infected with X4 or R5X4 clinical HIV isolates displaying the syncytium-inducing phenotype resulted in a complete suppression of X4 variants and a concomitant genotypic change in the V2 and V3 loops of the envelope gp120 glycoprotein. The recovered viruses corresponded genotypically and phenotypically to R5 variants in that they could no longer use CXCR4 as coreceptor or induce syncytium formation in MT-2 cells. Furthermore, the phenotype and genotype of a cloned R5 HIV-1 virus converted to those of the R5X4 virus after prolonged culture in lymphoid cells. However, these changes did not occur when the infected cells were cultured in the presence of AMD3100, despite low levels of virus replication. Our findings indicate that selective blockade of the CXCR4 receptor prevents the switch from the less pathogenic R5 HIV to the more pathogenic X4 HIV strains, a process that heralds the onset of AIDS. In this article, we show that it could be possible to redirect the evolution of HIV so as to impede the emergence of X4 strains or to change the phenotype of already-existing X4 isolates to R5. 相似文献
50.
Bonaventura Majolo Raffaella Ventura† & Nicola F. Koyama‡ 《Ethology : formerly Zeitschrift fur Tierpsychologie》2009,115(2):152-166
In various social species, animals have been observed to share friendly relationships with some group members and to resolve conflicts through reconciliation, the exchange of affiliative behaviour soon after a conflict that functions to restore the relationship between the former opponents. The valuable relationship hypothesis predicts that reconciliation should be observed more often after conflicts between friends. Friendly relationships can be described by three dimensions (i.e. value, security and compatibility); however, research into the relative importance of these dimensions for the occurrence of reconciliation is sparse. Moreover, reconciliation may depend on factors other than the social relationship between opponents including, for example, their social status or the context of the conflict. Our study aimed at analysing which factors are important determinants of reconciliation and at testing the valuable relationship hypothesis, by analysing the relative effects of relationship value, security and compatibility on the occurrence and timing of reconciliation. We collected data on two troops of wild Japanese macaques living on Yakushima Island, Japan, and selected the best predicting variables of reconciliation using linear mixed models. Our results show that reconciliation occurs more frequently, and earlier, after conflicts between opponents who exchange a higher percentage of grooming. Two additional variables related to relationship security and value were selected in the best models: frequency of aggression and of approaches resulting in tolerated co‐feeding. Among the variables not related to relationship quality, distance between opponents at the end of the conflict, kinship, sex of the opponents and context of conflict (i.e. during feeding or social time) were included in our models. Our findings support the valuable relationship hypothesis and, in particular, highlight that the fitness‐related benefits of social relationships (i.e. the relationship value) are important determinants of the evolution of friendly relationships and reconciliation. 相似文献