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81.
Pukala TL Urathamakul T Watt SJ Beck JL Jackway RJ Bowie JH 《Rapid communications in mass spectrometry : RCM》2008,22(22):3501-3509
Amphibian peptides which inhibit the formation of nitric oxide by neuronal nitric oxide synthase (nNOS) do so by binding to the protein cofactor, Ca2+calmodulin (Ca2+CaM). Complex formation between active peptides and Ca2+CaM has been demonstrated by negative ion electrospray ionisation mass spectrometry using an aqueous ammonium acetate buffer system. In all cases studied, the assemblies are formed with a 1:1:4 calmodulin/peptide/Ca2+ stoichiometry. In contrast, the complex involving the 20-residue binding domain of the plasma Ca2+ pump C20W (LRRGQILWFRGLNRIQTQIK-OH) with CaM has been shown by previous two-dimensional nuclear magnetic resonance (2D NMR) studies to involve complexation of the C-terminal end of CaM. Under identical conditions to those used for the amphibian peptide study, the ESI complex between C20W and CaM shows specific 1:1:2 stoichiometry. Since complex formation with the studied amphibian peptides requires Ca2+CaM to contain its full complement of four Ca2+ ions, this indicates that the amphibian peptides require both ends of the CaM to effect complex formation. Charge-state analysis and an H/D exchange experiment (with caerin 1.8) suggest that complexation involves Ca2+CaM undergoing a conformational change to a more compact structure. 相似文献
82.
83.
Watt SJ Urathamakul T Schaeffer PM Williams NK Sheil MM Dixon NE Beck JL 《Rapid communications in mass spectrometry : RCM》2007,21(2):132-140
The Escherichia coli DnaB protein (DnaB(6)) is the hexameric helicase that unwinds genomic DNA so it can be copied by the DNA replication machinery. Loading of the helicase onto DNA requires interactions of DnaB(6) with six molecules of its loading partner protein, DnaC. Nano-electrospray ionisation mass spectrometry (nanoESI-MS) of mutant proteins was used to examine the roles of the residues Phe102 (F102) and Asp82 (D82) in the N-terminal domain of DnaB in the assembly of the hexamer. When the proteins were prepared in 1 M ammonium acetate containing magnesium and adenosine triphosphate (ATP) at pH 7.6, both hexameric and heptameric forms of wild-type and F102W, F102E and D82N mutant DnaBs were observed in mass spectra. The spectra of the D82N mutant also showed substantial amounts of a decameric species and small amounts of a dodecamer. In contrast, the F102H DnaB mutant was incapable of forming oligomers of order higher than the hexamer. Thus, although Phe102 is not the only determinant of hexamer assembly, this residue has a role in oligomerisation. NanoESI mass spectra were obtained of mixtures of DnaB(6) with DnaC. The DnaB(6)(DnaC)(6) complex (calculated M(r) 481 164) was observed only when the two proteins were present in equimolar amounts. The data are consistent with cooperative assembly of the complex. ESI mass spectra of mixtures containing DnaC and ATP showed that DnaC slowly hydrolysed ATP to ADP as indicated by ions corresponding to DnaC/ATP and DnaC/ADP complexes. These experiments show that E. coli DnaB can form a heptameric complex and that nanoESI-MS can be used to probe assembly of large (>0.5 MDa) macromolecular complexes. 相似文献
84.
Salmanca Oviedo LN Gomez-Herrero A Landa Canovas AR Otero-Diaz LC 《Micron (Oxford, England : 1993)》2000,31(5):597-603
Two new misfit layer structures have been synthesized within the Sb-Nb-Se system. Powder X-ray diffraction and electron microscopy techniques (electron diffraction, HREM, XEDS) have been used to determine the nature of their structure. According to TEM and XEDS data (for more than 15 crystals studied) both phases are monolayer type, i.e. (SbSe)1+delta (NbSe2). Electron microscopy reveals a composite modulated structure that consists of the periodical intergrowth of a pseudotetragonal SbSe layer, denominated as Q, and a pseudohexagonal layer NbSe2, denominated as H. Both layers fit along b, stack along c and do not fit along a (misfit) giving rise to an incommensurate modulation along this direction. The two phases differ in the symmetry of the Q layers being in one case orthorhombic (for delta = 0.17) and monoclinic in the other (for delta = 0.19). After the characterization of the sample by electron microscopy the unit cells of the basic layers could be refined for both phases by powder X-ray diffraction: aQ = 5.824(2) A, bQ = 5.962(5) A, cQ = 23.927(6) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees and aH = 3.415(5) A, bH = 5.962(6) A,, cH = 11.962(1) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees for the orthorhombic phase; aQ = 5.844(2) A, bQ = 5.981(1) A, cQ = 23.919(5) A, alpha = 90 degrees, beta = 90 degrees and gamma = 96.00(3)degrees and aH = 3.439(1) A, bH = 5.994(2) A, cH = 11.956(3) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees for the monoclinic phase. The phase with the monoclinic Q-sublattice often appears as twinned crystals. The more abundant crystals are disordered intergrowths of both monolayer phases. 相似文献
85.
Bezuidenhout DI Barnard W van der Westhuizen B van der Watt E Liles DC 《Dalton transactions (Cambridge, England : 2003)》2011,40(25):6711-6721
Fischer carbene complexes of tungsten with substituents containing up to two additional different transition metals, with all the metals in electronic contact with the carbene carbon atom, were synthesised and studied both in solution and in the solid state. For the complexes of the type [W(CO)(5){C(OR')R}], the substituents chosen were heteroaromatic 2-benzo[b]thienyl (2-BT), or 2-BT π-bonded to a chromium tricarbonyl fragment ([Cr(CO)(3)(2-η(6)-BT)]) or ferrocenyl (Fc) as the R-substituent, while the OR'-substituent was systematically varied between an ethoxy or a titanoxy group, to yield the complexes 1b (R' = Et, R = 2-BT), 2b (R' = Et, R = [Cr(CO)(3)(2-η(6)-BT)]), 3b (R' = TiCp(2)Cl, R = 21-BT), 4b (R' = TiCp(2)Cl, R = [Cr(CO)(3)(2-η(6)-BT)]), 5b (R' = Et, R = Fc) and 6b (R' = TiCp(2)Cl, R = Fc). The structural features and their relevance to bonding in the multimetal carbene compounds of both these tungsten and the analogous chromium complexes were investigated as they represent indicators of possible reactivity sites in multimetal carbene assemblies. The possibility of using DFT calculations to quantify the effect of metal-containing substituents on the carbene ligands was tested and correlated with experimental parameters by employing methods such as vibrational spectroscopy, molecular orbital analysis, and cyclic voltammetry. 相似文献
86.
Orihuela R Fernández B Palacios O Valero E Atrian S Watt RK Domínguez-Vera JM Capdevila M 《Chemical communications (Cambridge, England)》2011,47(44):12155-12157
Ferritin (Ft) interaction with the Zn-complexes of mammalian MT1, MT2 and MT3 metallothioneins (MT) leads to simultaneous Fe(II) and Zn(II) release. 相似文献
87.
López-Castro JD Delgado JJ Perez-Omil JA Gálvez N Cuesta R Watt RK Domínguez-Vera JM 《Dalton transactions (Cambridge, England : 2003)》2012,41(4):1320-1324
An electron microscopy study, in combination with modeling and image simulation, of four different reconstituted ferritin samples: recombinant human H and L homopolymers, and H and L heteropolymers of native L-subunit-rich horse spleen and H-subunit-rich human heart ferritins, points out the existence of a correlation between iron core shape and protein shell. 相似文献
88.
Sulaf Assi Robert A. Watt Anthony C. Moffat 《Journal of Raman spectroscopy : JRS》2012,43(8):1049-1057
Quantification of ciprofloxacin in proprietary Ciproxin tablets and generic ciprofloxacin tablets was made using handheld Raman spectroscopy. Powder dilutions of ciprofloxacin or crushed Ciproxin tablets with excipient(s) were made. The spectra were taken either from the powder dilutions or from pellets compressed from these dilutions and were stored in the instrument library. Methods were created from these powder dilutions: each representing a separate model. The spectra of the tablets were identified within each method using the inbuilt algorithm of the instrument. The results showed that only the models made with the crushed Ciproxin tablets were semi‐quantitative for the analysis of Ciproxin tablets. On the other hand, none of the models could identify generic ciprofloxacin tablets. Thus, the spectra were exported to MATLAB for off‐line quantification. Whereas, univariate regression models were hardly linear, partial least square regression models gave linear calibrations with correlation coefficients above 0.99 and root mean square error of calibration below 3.75% m/m. For the prediction of tablets, the models varied in their results with the one created with ciprofloxacin and lactose giving the most satisfactory results. However, in total the method did not give the pharmacopoeial accuracy of ±5% and stays a semi‐quantitative approach although a satisfactory identity of the constituents was achieved. Copyright © 2011 John Wiley & Sons, Ltd. 相似文献
89.
High-pulse-energy supercontinuum radiation with a width exceeding 900 nm in the near-infrared spectral region has been generated
in conventional single-mode fibre. The fibre was pumped at 1064 nm which is in the normal dispersion regime, resulting in
predominantly red-shifted spectral broadening. Supercontinuum pulse energies exceeding 450 nJ were obtained. The use of conventional
fibre allows for inexpensive generation of near-infrared supercontinuum radiation, featuring high pulse energies and good
spatial beam quality. This supercontinuum radiation was used to acquire high-resolution (15 pm) broadband absorption spectra
of H2O, C2H2 and C2H4 in the near-infrared spectral region (1340–1700 nm), using an optical spectrum analyser for detection. H2O spectra were also recorded at high repetition rates, by dispersing the supercontinuum pulses and detecting the transmitted
signal in the time domain. A spectral resolution of 38 pm was obtained employing the dispersed supercontinuum pulses, which
is comparable to the H2O line widths at ambient conditions.
PACS 07.07.Df; 42.62.Fi; 42.79.Nv; 42.81.-i 相似文献
90.