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1.
Villalba J. M. Navarro F. Roldán J. M. González-Reyes J. A. Navas P. 《Protoplasma》1994,178(3-4):87-96
Summary Expression of various sugar residues on the plasma membrane of frog (Rana perezi) epidermal cells at different stages of differentiation has been monitored with the use of a battery of HRP-conjugated lectins. In paraffin-embedded tissue, mannose residues (stained by Concanavalin A) were detected at the keratinocyte cell surface in all epidermal strata. However,Lens culinaris agglutinin (LCA), also specific for mannose, specifically stained the plasma membrane of cells from the stratum germinativum. Expression of N-acetyl-glucosamine (GlcNAc), labelled with wheat germ agglutinin (WGA), was maximum at the cell surface of basal cells and progressively decreased through the stratum spinosum. Galactose (Gal) and N-acetyl-galactosamine (GalNAc) residues, labelled withGriffonia simplicifolia I (GS I) andGlycine max (SBA) agglutinins, respectively, were expressed according to the degree of differentiation in amphibian epidermal cells. Sialic acid-containing glycoproteins, labelled withLimax flavus agglutinin (LFA), were found in the outermost plasma membrane of the replacement cell layer and stratum corneum. Glycoproteins responsible for the observed lectin-binding patterns have been identified by staining on nitrocellulose filters after electrophoresis of solubilized plasma membrane fractions and Western blotting. Changes at the level of glycosylation of plasma membrane glycoproteins as epidermal cells differentiate are discussed on the basis of a progressive addition of Gal residues. Integral membrane proteins have been solubilized with the non-denaturing detergent CHAPS and glycoproteins containing terminal Gal residues, that are expressed according to the degree of differentiation in frog epidermis, have been partially purified by affinity chromatography on a GS I-Sepharose 4 B column. The purified fraction was composed by four acidic glycoproteins with isoelectric points between 4.6 and 5.2 and, in SDS-gels gave five major protein bands with approximate molecular weights of 148, 140, 102, 60, and 52 kDa in SDS-gels. The 102 and 52 kDa bands correspond to the a and subunits of amphibian epidermal Na+,K+-ATPase as demonstrated by specific staining with a polyclonal antibody against the catalytic subunit of pig kidney proton pump and staining with lectins GS I, GS II, and WGA. Possible relationships between higher molecular weight proteins and the constituents of intramembranous particles from the outermost plasma membranes of the replacement cell layer and the stratum corneum are also discussed.Abbreviations BSA
bovine serum albumin
- CHAPS
(3-[(cholamidopropyl) dimethyl-ammonio] 1-propanesulfonate)
- Con A
Canavalia ensiformis agglutinin
- DTT
dithiothreitol
- Gal
galactose
- GalNAc
N-acetyl-D-galactosamine
- GlcNAc
N-acetyl-D-glucosamine
- GS I
Griffonia simplicifolia agglutinin I
- GS II
Griffonia simplicifolia agglutinin II
- HRP
horseradish peroxidase
- LFA
Limax flavus agglutinin
- LCA
Lens culinaris agglutinin
- NDPAGIF
non-denaturing polyacrylamide gel isoelectric focusing
- PAGE
polyacrylamide gel electrophoresis
- PAP
peroxidase-antiperoxidase
- PBS
phosphate buffered saline
- PMSF
phenyl methyl sulphonyl fluoride
- RCL
replacement cell layer
- SBA
soybean agglutinin (Glycine max)
- SB
stratum basal
- SDS
sodium dodecyl sulphate
- SG
stratum granulosum
- SS
stratum spinosum
- UEA I
Ulex europaeus agglutinin I
- WGA
wheat germ (Triticum vulgaris) agglutinin 相似文献
2.
J. C. Rodríguez-Aguilera F. Navarro A. Arroyo F. J. Alcaín J. M. Villalba P. Navas 《Protoplasma》1995,184(1-4):229-232
Summary Ascorbate is stabilized in the presence of HL-60 cells. Our results showed that cAMP derivatives and agents that increase cAMP stimulate the ability of HL-60 cells to stabilize ascorbate. On the other hand, tunicamycin, a glycosilation-interfering agent, inhibited this ability. The ascorbate stabilization in the presence of HL-60 cells has been questioned as a simple chemical effect. Further properties and controls about the enzymatic nature of this stabilization are described and discussed. This data, together with hormonal regulation, support the hypothesis that an enzymatic redox system located at the plasma membrane is responsible of the extracellular ascorbate stabilization by HL-60 cells.Abbreviations AFR
ascorbate free radicals
- FCS
fetal calf serum
- Sp-cAMPS
Sp-cyclic adenosine monophosphothionate
- Rp-cAMPS
Rp-cyclic adenosine monophosphothionate 相似文献
3.
A structural and histochemical study of the tongue in the Anuran Amphibian Rana ridibunda was carried out. Histochemical analysis of the filiform and fungiform papillae of the dorsal epithelium has shown a variety of cellular types which may be characterized cytochemically. All of them, except the goblet cells, show a remarkable amount of neutral mucins. The intensity of histochemical positive reaction for sulphomucins, sialomucins and protein is variable according to the cell type. Other histochemical reactions show that the lingual glands are rich in neutral mucins, but not in sialomucins. Histochemically in the component the basic proteins with sulphydryl groups are demonstrated. This sulphydryl groups are more abundant in the glands located in the deep regions. 相似文献
4.
P Navas I L Sun D J Morré F L Crane 《Biochemical and biophysical research communications》1986,135(1):110-115
The short-term incubation of HeLa cells in the presence of diferric transferrin or ferricyanide, which are reduced externally by the transplasma membrane reductase, produces a stoichiometric decrease in NADH and increase in NAD+, which is stimulated by insulin. The NADP/NADPH ratio does not change during 15 min incubation with the oxidants. The total pyridine nucleotide pool of HeLa cells is not affected. Incubation with apotransferrin and ferrocyanide, which cannot act as oxidants for transmembrane electron transport, does not change the pyridine nucleotide concentrations in the cells. Our results show that NADH can act as the internal electron donor for the reduction of external oxidants by the transmembrane reductase. It appears that oxidation of NADH by the transmembrane electron transport using ferricyanide or iron transferrin as external electron acceptors is sufficient to stimulate growth in HeLa cells. 相似文献
5.
Yana Chen Kevork Hagopian Douglas Bibus José?M. Villalba Guillermo López-Lluch Plácido Navas Kyoungmi Kim Roger?B. McDonald Jon?J. Ramsey 《Bioscience reports》2013,33(1)
To investigate the role mitochondrial membrane lipids play in the actions of CR (calorie restriction), C57BL/6 mice were assigned to four groups (control and three 40% CR groups) and the CR groups were fed diets containing soya bean oil (also in the control diet), fish oil or lard. The fatty acid composition of the major mitochondrial phospholipid classes, proton leak and H2O2 production were measured in liver mitochondria following 1 month of CR. The results indicate that mitochondrial phospholipid fatty acids reflect the PUFA (polyunsaturated fatty acid) profile of the dietary lipid sources. CR significantly decreased the capacity of ROS (reactive oxygen species) production by Complex III but did not markedly alter proton leak and ETC (electron transport chain) enzyme activities. Within the CR regimens, the CR-fish group had decreased ROS production by both Complexes I and III, and increased proton leak when compared with the other CR groups. The CR-lard group showed the lowest proton leak compared with the other CR groups. The ETC enzyme activity measurements in the CR regimens showed that Complex I activity was decreased in both the CR-fish and CR-lard groups. Moreover, the CR-fish group also had lower Complex II activity compared with the other CR groups. These results indicate that dietary lipid composition does influence liver mitochondrial phospholipid composition, ROS production, proton leak and ETC enzyme activities in CR animals. 相似文献
6.
Sergio Padilla-López María Jiménez-Hidalgo Alejandro Martín-Montalvo Catherine F. Clarke Plácido Navas Carlos Santos-Ocaña 《生物化学与生物物理学报:生物膜》2009,1788(6):1238-1248
Coenzyme Q is an isoprenylated benzoquinone lipid that functions in respiratory electron transport and as a lipid antioxidant. Dietary supplementation with Q is increasingly used as a therapeutic for treatment of mitochondrial and neurodegenerative diseases, yet little is known regarding the mechanism of its uptake. As opposed to other yeast backgrounds, EG103 strains are unable to import exogenous Q6 to the mitochondria. Furthermore, the distribution of exogenous Q6 among endomembranes suggests an impairment of the membrane traffic at the level of the endocytic pathway. This fact was confirmed after the detection of defects in the incorporation of FM4-64 marker and CPY delivery to the vacuole. A similar effect was demonstrated in double mutant strains in Q6 synthesis and several steps of endocytic process; those cells are unable to uptake exogenous Q6 to the mitochondria and restore the growth on non-fermentable carbon sources. Additional data about the positive effect of peptone presence for exogenous Q6 uptake support the hypothesis that Q6 is transported to mitochondria through an endocytic-based system. 相似文献
7.
Carvalho JE Gomes FR Navas CA 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2008,178(4):447-456
Anuran amphibians exhibit different patterns of energy substrate utilization that correlate with the intensity of vocal and
locomotor activities. Given the remarkable differences among species in breeding and feeding strategies, and the different
ways energy is used in the whole animal, the suggested correlations between calling and locomotor behavior and the level of
energy substrates in the muscles responsible for such activities are more complex than previously reported. We explored the
relationships between calling and locomotor behavior and energy supply to trunk and hindlimb muscles, respectively, within
the ecologically diverse tree-frog genus Scinax. Specifically, we measured the relative amount of carbohydrates and lipids in these two groups of muscles, and in the liver
of three species of Scinax that differ in vocal and locomotor performance, and compared our results with those of two other species for which comparable
data are available. We also compared the contents of lipids and carbohydrates of conspecific males collected at the beginning
and after 4 h of calling activity. The stomach content to potential feeding opportunities across species was also assessed
in both groups of males. Scinax hiemalis and S. rizibilis exhibit comparatively low and episodic calling during long periods of activity whereas S. crospedospilus calls at higher rates over shorter periods. Male S. hiemalis had highest levels of trunk muscle glycogen followed by those of S. rizilbilis and S. crospedospilus, respectively. There was no correlation between total lipid content in trunk muscle and calling rate among different species,
suggesting that other metabolic aspects may be responsible for the energetic support for vocal activity. The levels of lipids
and carbohydrates in trunk and hindlimb muscles and liver of males collected at the beginning and 4 h into the calling period
were similar across species, so the extent of energetic reserves does not appear to constrain vocal or locomotor activity.
Finally, we found exceptionally high levels of carbohydrates and lipids in the liver of S. rizibilis, a trait perhaps related to a long and demanding breeding period. 相似文献
8.
9.
Panizo María Mercedes Ferrara Giuseppe García Nataly Moreno Xiomara Navas Trina Calderón Enrique 《Current fungal infection reports》2020,14(1):29-39
Current Fungal Infection Reports - The aim of this work is to contribute to the knowledge of diagnosis, burden, and mortality of pneumocystosis or Pneumocystis jirovecii pneumonia (PCP) in... 相似文献
10.
Helen S. Findlay Yuri Artioli Juan Moreno Navas Sebastian J. Hennige Laura C. Wicks Veerle A. I. Huvenne E. Malcolm S. Woodward J. Murray Roberts 《Global Change Biology》2013,19(9):2708-2719
Cold‐water coral (CWC) reefs are recognized as ecologically and biologically significant areas that generate habitats and diversity. The interaction between hydrodynamics and CWCs has been well studied at the Mingulay Reef Complex, a relatively shallow area of reefs found on the continental shelf off Scotland, UK. Within ‘Mingulay Area 01’ a rapid tidal downwelling of surface waters, brought about as an internal wave, is known to supply warmer, phytoplankton‐rich waters to corals growing on the northern flank of an east‐west trending seabed ridge. This study shows that this tidal downwelling also causes short‐term perturbations in the inorganic carbon (CT) and nutrient dynamics through the water column and immediately above the reef. Over a 14 h period, corresponding to one semi‐diurnal tidal cycle, seawater pH overlying the reef varied by ca. 0.1 pH unit, while pCO2 shifted by >60 μatm, a shift equivalent to a ca. 25 year jump into the future, with respect to atmospheric pCO2. During the summer stratified period, these downwelling events result in the reef being washed over with surface water that has higher pH, is warmer, nutrient depleted, but rich in phytoplankton‐derived particles compared to the deeper waters in which the corals sit. Empirical observations, together with outputs from the European Regional Shelf Sea Ecosystem Model, demonstrate that the variability that the CWC reefs experience changes through the seasons and into the future. Hence, as ocean acidification and warming increase into the future, the downwelling event specific to this site could provide short‐term amelioration of corrosive conditions at certain times of the year; however, it could additionally result in enhanced detrimental impacts of warming on CWCs. Natural variability in the CT and nutrient conditions, as well as local hydrodynamic regimes, must be accounted for in any future predictions concerning the responses of marine ecosystems to climate change. 相似文献