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71.
Kaznacheyeva E Zubov A Nikolaev A Alexeenko V Bezprozvanny I Mozhayeva GN 《The Journal of biological chemistry》2000,275(7):4561-4564
In most nonexcitable cells, calcium (Ca(2+)) release from inositol 1,4,5-trisphosphate (InsP(3))-sensitive intracellular Ca(2+) stores is coupled to Ca(2+) influx (calcium release-activated channels (I(CRAC))) pathway. Despite intense investigation, the molecular identity of I(CRAC) and the mechanism of its activation remain poorly understood. InsP(3)-dependent miniature calcium channels (I(min)) display functional properties characteristic for I(CRAC). Here we used patch clamp recordings of I(min) channels in human carcinoma A431 cells to demonstrate that I(min) activity was greatly enchanced in the presence of anti-phosphatidylinositol 4, 5-bisphosphate antibody (PIP(2)Ab) and diminished in the presence of PIP(2). Anti-PIP(2) antibody induced a greater than 6-fold increase in I(min) sensitivity for InsP(3) activation and an almost 4-fold change in I(min) maximal open probability. The addition of exogenous PIP(2) vesicles to the cytosolic surface of inside-out patches inhibited I(min) activity. These results lead us to propose an existence of a Ca(2+) influx pathway in nonexcitable cells activated via direct conformational coupling with a selected population of InsP(3) receptors, located just underneath the plasma membrane and coupled to PIP(2). The described pathway provides for a highly compartmentalized Ca(2+) influx and intracellular Ca(2+) store refilling mechanism. 相似文献
72.
Fedorova OA Gromov SP Eshcheulova OV Zeiss V Moebius D Baryshnikova EA Vereshchetin VP Sergeeva TI Zubov VP Zaĭtsev SIu Alfimov MV 《Bioorganicheskaia khimiia》2000,26(9):707-717
New amphiphilic photochromic benzo-15(18)-crown-5(6) ethers (APC) differing in the position of the octadecyl substituent and the size of the crown cavity were synthesized. The compounds form stable monolayers in the air/water and air/alkaline metal salt solution interfaces. The results of the pressure isotherm measurements, atomic force microscopy (AFM), and electronic spectroscopy show that the structure of the monolayers formed depends on the structure of the parent APC and the nature of the cation in salt solutions. The area per molecule of APC in the monolayer (specific area) is the smallest on the water surface and increases by 20-40% on the aqueous subphase surface with an increasing concentration of salts therein to indicate the formation of APC complexes with the metal cations. When the hydrophobic aliphatic substituent is displaced from position 3 to position 5 of the benzothiazole ring, the specific area on the surface of water and subphases decreases twofold, which indicates the compactization of the monolayer on this modification. A reversible E-Z-photoisomerization of APC was found in the monolayers formed in the salt solution/air interface. The features of the reaction are defined by the specific organization of the amphiphilic molecules in the monolayer and by the nature of the cation. 相似文献
73.
74.
E P Erokhin I S Tartakovski? O V Radchenko Iu V Lukin D M Avdeev N K Misurenko V P Zubov S V Prozorovski? 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1991,(11):41-43
The method of synthetizing dispersions of gelatin-modified polyacrolein microspheres 1.5-2.5 micrograms in diameter, used as a solid-phase carrier for the preparation of immunodispersion diagnostica, has been developed. The possibility of using immunodispersion diagnosticum in the passive agglutination test with hyperimmune rabbit sera has been demonstrated. High activity and specificity of immunodispersion diagnosticum, combining methodological simplicity and rapidity characteristic of agglutination, has been shown. 相似文献
75.
Ionic currents of cells of neuroblastoma clone N18 A-1 was studied under conditions when the internal medium was placed for artificial fluoride or phosphate solutions. The specific membrane leakage resistance was measured to be 8.1 +/- 2.6 kOhm.cm2 and 1.3 +/- 0.3 Kohm.cm2, respectively. The presence of usual sodium and tetraethylammonium sensitive potassium channels is demonstrated. Potassium conductance is shown to amount to 0.25--0.025 of sodium conductance. Dialysis of the cells by phosphate solutions induces a slow outward current, which is not inhibited by tetraethylammonium ions. 相似文献
76.
L V Kozlov B B Sho?bonov A E Ivanov V P Zubov V K Antonov 《Biokhimii?a (Moscow, Russia)》1989,54(10):1745-1751
An affinity sorbent comprising macroporous glass coated with the polymer with the polymer with immobilized immunoglobulin IgG was used for the isolation from human serum of the first component of the complement and for its separation into subcomponents C1r, C1s and C1q by the one-step procedure. Serum C1 was quantitatively bound to the sorbent at 0 degrees C. The unbound part of the serum can be used as a R1 reagent for determining the hemolytic activity of C1. After activation of bound C1 by heating (30 degrees C, 40 min) the activated subcomponent C1r is eluted from the sorbent. Stepwise elution with EDTA at pH 7.4 or with EDTA + 1 M NaCl at pH 8.5 results in a selective and quantitative elution of the activated subcomponent C1s and subcomponent C1q. Stepwise elution of C1 subcomponents from the affinity sorbent after activation reflects the process of C1 breakdown following its activation on immune complexes. 相似文献
77.
Vikhrov A.A. Markvicheva E.A. Mareeva T.Yu. Khaidukov S.V. Nesmeyanov V.A. Manakov M.N. Goergen J-L. Marc A. Zubov V.P. 《Biotechnology Techniques》1998,12(1):11-14
A simple and reliable technique was developed to prepare pure monoclonal antibody (MAb) to interleukin-2 using cells entrapped in novel composite poly(N-vinyl caprolactam)-calcium alginate beads. Flow cytometry was applied to study cell size and cell cytoplasm granularity distribution. Maximum MAb production by the gel-entrapped cells in serum free medium was 2-3-fold higher compared to free suspension culture in serum containing medium. The only contaminating protein in culture supernatant was transferrin at 5% w/v. 相似文献
78.
79.
Marina V. Donova Irina F. Kuzkina Anna Yu. Arinbasarova Igor I. Pashkin Elena A. Markvicheva Tatyana G. Baklashova Galina V. Sukhodolskaya Victoria V. Fokina Yurii E. Kirsh Kira A. Koshcheyenko Vitalii P. Zubov 《Biotechnology Techniques》1993,7(6):415-422
Summary A new gel-type support poly-N-vinylcaprolactam for microbial cell immobilization is presented. The method allows one to obtain beads of biocatalyst in a single step. The properties of beads obtained using different types of gel stabilizers were compared; the best stabilizer was found to be tannin. The method developed was used for entrapment of viable bacterial cells and fungal spores. The biocatalysts obtained were used for transformations of both hydrophilic (sorbitol, indolyl-3-acetic acid) and lipophilic (cortexolone, hydrocortisone) substrates.Abbreviations PVC
poly-N-vinylcaprolactam
- ImC
immobilized cells
- IAA
indolyl-3-acetic acid
- TLC
thin layer chromatography 相似文献
80.
K. S. Stashevskaya E. A. Markvicheva S. M. Strukova A. V. Rusanova A. M. Makarova L. R. Gorbacheva I. A. Prudchenko V. P. Zubov K. Grandfis 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2007,1(2):147-154
Thrombin receptor agonist peptide (TRAP-6) may be successfully used instead of thrombin to stimulate regeneration of damaged tissues. Thrombin application is limited by its high price, instability, and proin-flammatory effect at high concentrations. Immobilization of TRAP-6 into a matrix based on lactic and glycolic acid copolymer (PLGA) prevents its destruction by peptidases located in the wound and can also provide controlled release of the peptide. PLGA microparticles with the immobilized peptide were prepared by the double emulgation method. The presence of the immobilized peptide increased the porosity of the microparticle surface detected by scanning electron microscopy. Kinetics of the TRAP-6 release was characterized by a dramatic increase in its concentration in buffer solution (pH 7.5) during the first 2 h after the experiment beginning, and the complete release of the peptide after 20 h. An investigation of TRAP-6 destruction by scanning electron microscopy revealed the increase in the microparticle size and surface porosity already after one day of incubation, and the destroyed microparticles were aggregated by the seventh day of the incubation. Thus, peptide immobilization into PLGA microparticles may be employed for elaboration of a prolonged action preparation with the controlled release of the active agent (peptide). 相似文献