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101.
Koichi Iwata Masahide Terazima Hiroshi Masuhara 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(2):335-357
Novel methodologies utilizing pulsed or intense CW irradiation obtained from lasers have a major impact on biological sciences. In this article, recent development in biophysical researches fully utilizing the laser irradiation is described for three topics, time-resolved fluorescence spectroscopy, time-resolved thermodynamics, and manipulation of the biological assemblies by intense laser irradiation. First, experimental techniques for time-resolved fluorescence spectroscopy are concisely explained in Section 2. As an example of the recent application of time-resolved fluorescence spectroscopy to biological systems, evaluation of the viscosity of lipid bilayer membranes is described. The results of the spectroscopic experiments strongly suggest the presence of heterogeneous membrane structure with two different viscosity values in liposomes formed by a single phospholipid. Section 3 covers the time-resolved thermodynamics. Thermodynamical properties are important to characterize biomolecules. However, measurement of these quantities for short-lived intermediate species has been impossible by traditional thermodynamical techniques. Recently, development of a spectroscopic method based on the transient grating method enables us to measure these quantities and also to elucidate reaction kinetics which cannot be detected by other spectroscopic methods. The principle of the measurements and applications to some protein reactions are reviewed. Manipulation and fabrication of supramolecues, amino acids, proteins, and living cells by intense laser irradiation are described in Section 4. Unconventional assembly, crystallization and growth, amyloid fibril formation, and living cell manipulation are achieved by CW laser trapping and femtosecond laser-induced cavitation bubbling. Their spatio-temporal controllability is opening a new avenue in the relevant molecular and bioscience research fields. This article is part of a Special Issue entitled “Biophysical Exploration of Dynamical Ordering of Biomolecular Systems” edited by Dr. Koichi Kato. 相似文献
102.
Kondo Masahide Taya Toshiki Matsuba Takao Fushimi Noriko Inouye Kuniyo Kidokoro Shun-ichi Yasukawa Kiyoshi 《Biotechnology Techniques》1996,10(8):547-552
Summary By using a commercially available surface plasmon resonance (SPR) biosensor, the values of the association rate constant (kass), dissociation rate constant (kdiss), and association constant (KA = kass / kdiss) for binding to the antigens were determined. They were almost the same for the recombinant antibody expressed in COS cells, CHO cells, and mouse hybridoma cells. The system of transient expression of the recombinant antibody (Ab) in COS cells and SPR analysis of the supernatant should be useful for rapid expression and evaluation of the binding ability of large numbers of engineered Abs. 相似文献
103.
We have developed a new immunostaining technique specifically for the detection of human tumors transplanted into nude mice
using mouse monoclonal antibodies (MoAbs) produced in our laboratory. The formation of a molecular complex consisting of three
components (mouse MoAb or hybridoma supernatant, biotin-labeled anti-mouse immunoglobulins, and normal nude mouse serum) markedly
reduced background staining and enhanced specific reaction with the transplanted tumors in nude mice. When MoAb production
by electrofusion was screened with this new method, the incidence of hybridoma supernatant reactive with sections of transplanted
tumor was 2.3 per 100 wells immunostained. These results suggest that production of MoAbs using transplanted tumors is immunohistochemically
possible and that this method may provide a new means for developing useful tumor markers.
Accepted: 30 September 1996 相似文献
104.
105.
Masanobu Tokushige Katsumi Iinuma Masahide Yamamoto Yasunori Nishijima 《Biochemical and biophysical research communications》1980,96(2):863-869
Excitation of apotryptophanase from Escherichia coli at 290 nm yielded a fluorescence emission centered at 340 nm. Binding of pyridoxal phosphate to apoenzyme induced quenching of protein fluorescence concomitant with an appearance of another peak at 510 nm by way of energy transfer from tryptophan. Based on the results, an approximate distance between the coenzyme and tryptophan was estimated to be 18–24 Å according to the Förster's theory. The ozone-inactivated enzyme yielded only the 340 nm-peak upon excitation at 290 nm following reconstitution with the coenzyme. The fluorescence decay time of the tryptophyl residue was somewhat increased by ozone-inactivation. These results suggest that the tryptophyl residue essential for the activity is involved in a direct interaction with the coenzyme. 相似文献
106.
Masahide Ishibashi 《Journal of bacteriology》1967,93(1):379-389
Specific aggregate formation of F pili was observed, by electron microscopy, in a mixture of male Escherichia coli (or of isolated F pili) and anti-f(+) serum. Cellular appendages other than F pili never showed such aggregation when mixed with anti-f(+) serum. The f(+) agglutinability of male cells, as well as F piliation, was sensitive to mechanical agitation. The f(+) agglutination was inhibited when appropriate numbers of phage M12, capable of attaching to F pili, were mixed with the male culture before the addition of anti-f(+) serum. Correlation between f(+) agglutinability and the extent of F piliation was observed. It was concluded that the F pilus is the structure of the f(+) antigen and is responsible for f(+) agglutination. 相似文献
107.
Shingo Kiyokawa Yoshiyuki Hirata Yasuo Nagaoka Makio Shibano Masahiko Taniguchi Masahide Yasuda Kimiye Baba Shinichi Uesato 《Bioorganic & medicinal chemistry》2010,18(11):3925-3933
New 2-aminobenzamide-type histone deacetylase (HDAC) inhibitors were synthesized. They feature a sulfur-containing bicyclic arylmethyl moiety—a surface recognition domain introduced to increase in cellular uptake—and a substituted tert-amino group which affects physicochemical properties such as aqueous solubility. Compound 22 with a (2-hydroxyethyl)(4-(thiophen-2-yl)benzyl)amino group reduced the volume of human colon cancer HCT116 xenografts in nude mice to T/C 67% by oral administration at 45 mg/kg, which was comparable to the rate (T/C 62%) for a positive control, MS-275. Western blot analyses as well as cell cycle and TUNEL assays by flow cytometry suggested that the two compounds inhibited the growth of cancer cells via similar mechanisms. 相似文献
108.
109.
Koichiro Fukuoka Fuminori Suda Ryo Suzuki Masahide Ishikawa Hiroshi Takaku Tsujiaki Hata 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1557-1567
Abstract Bifunctional phosphorylating reagents 1 and 2 were employed for the synthesis of the cap part, m7G7 pppG, from guanosine 5′-phosphates on a large scale without any protecting groups. 相似文献
110.
The nexin-dynein regulatory complex (N-DRC) forms a cross-bridge between the outer doublet microtubules of the axoneme and regulates dynein motor activity in cilia/flagella. Although the molecular composition and the three-dimensional structure of N-DRC have been studied using mutant strains lacking N-DRC subunits, more accurate approaches are necessary to characterize the structure and function of N-DRC. In this study, we precisely localized DRC1, DRC2, and DRC4 using cryo–electron tomography and structural labeling. All three N-DRC subunits had elongated conformations and spanned the length of N-DRC. Furthermore, we purified N-DRC and characterized its microtubule-binding properties. Purified N-DRC bound to the microtubule and partially inhibited microtubule sliding driven by the outer dynein arms (ODAs). Of interest, microtubule sliding was observed even in the presence of fourfold molar excess of N-DRC relative to ODA. These results provide insights into the role of N-DRC in generating the beating motions of cilia/flagella. 相似文献