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排序方式: 共有104条查询结果,搜索用时 15 毫秒
1.
Binding characteristics of reduced hepatic receptors for acetylated low-density lipoprotein and maleylated bovine serum albumin. 总被引:3,自引:0,他引:3
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E Ottnad D P Via H Sinn E Friedrich R Ziegler H A Dresel 《The Biochemical journal》1990,265(3):689-698
The binding characteristics of reduced hepatic membrane proteins for acetylated low-density lipoprotein (acetyl-LDL) and maleylated bovine serum albumin (Mal-BSA) have been examined. Two receptor activities were extracted from hepatic membranes in the presence of octyl beta-D-glucoside and beta-mercaptoethanol, and were separated by chromatography on Mal-BSA-Sepharose 4B. The receptors were revealed by ligand blotting. The active binding proteins had apparent molecular masses of 35 and 15 kDa in SDS/polyacrylamide gels. Equilibrium studies with protein-phosphatidylcholine complexes indicated that the reduced 35 kDa protein expresses two binding sites for Mal-BSA and one for acetyl-LDL, whereas the 15 kDa protein-phosphatidylcholine complex binds 131I-Mal-BSA and 131I-acetyl-LDL with a 4:1 stoichiometry. 131I-Mal-BSA binding was linear with both proteins, with a Kd of 4.8 nM at the 35 kDa protein and a Kd of 5.6 nM at the 15 kDa protein. The 35 kDa protein displayed saturable binding of 131I-acetyl-LDL with a Kd of 5 nM; the 15 kDa binding protein bound 131I-acetyl-LDL with a Kd of 2.3 nM. A 85 kDa protein was obtained by Mal-BSA-Sepharose chromatography when the hepatic membranes had been solubilized with Triton X-100 in presence of GSH/GSSG. This protein displayed saturable 131I-Mal-BSA binding with a Kd of 30 nM and 131I-acetyl-LDL binding with a Kd of 6.5 nM. The 131I-Mal-BSA binding capacity was four times higher than that of 131I-acetyl-LDL. Competition studies with the 35 kDa, 15 kDa and 85 kDa proteins binding Mal-BSA, acetyl-LDL, formylated albumin and polyanionic competitors provide evidence for the existence of more than one class of binding sites at the reduced binding proteins. 相似文献
2.
Regulation of heparan sulphate metabolism by adenosine 3′:5′-cyclic monophosphate in hepatocytes in culture
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Freshly isolated rat hepatocytes maintained as monolayers in a serum-free medium synthesize sulphated glycosaminoglycans, most of which behave as heparan sulphate and are mainly distributed into intracellular compartments. Cyclic AMP, dibutyryl cyclic AMP, glucagon, noradrenaline, prostaglandin E(1), and theophylline, all drugs and hormones known to increase intracellular cyclic AMP concentrations, decreased the incorporation of (35)SO(4) (2-) into heparan sulphate of intra-, extra- and peri-cellular pools. The inhibition mediated by dibutyryl cyclic AMP was dose-dependent and observed as early as 2h after exposure to the drug. In the presence of 1mm-dibutyryl cyclic AMP, incorporation of (35)SO(4) (2-) or [(14)C]glucosamine into heparan sulphate was decreased to 40-50%, suggesting that dibutyryl cyclic AMP interfered with the synthesis of heparan sulphate. This was further supported by pulse-chase experiments, where dibutyryl cyclic AMP had no effect on the degradation of sulphated glycosaminoglycans. Heparan sulphates synthesized and secreted into the extracellular pool in the presence of dibutyryl cyclic AMP were smaller in size, whereas the degree of sulphation and molecular size of the heparan sulphate chains released by beta-elimination from these proteoglycans were not different from control values. In the presence of 1mm-cycloheximide, (35)SO(4) (2-) incorporation was decreased to 5%. Addition of p-nitrophenyl beta-d-xyloside, an artificial acceptor of glycosaminoglycan chain synthesis, enhanced this incorporation to 18%. Dibutyryl cyclic AMP did not have any inhibitory effect on the synthesis of chains initiated on p-nitrophenyl beta-d-xylosides. Incorporation of [(3)H]serine into heparan sulphate was not affected by dibutyryl cyclic AMP, whereas the degree of substitution of serine residues with heparan sulphate chains was less in heparan sulphate synthesized in the presence of dibutyryl cyclic AMP, suggesting that cyclic AMP exerts its effect on the metabolism of sulphated glycosaminoglycans by affecting the transfer of xylose on to the protein core. 相似文献
3.
The secretion of heparan sulphate by cultured rat hepatocytes was increased in the presence of (+)-catechin. The increase was due to a new species of heparan sulphate that lacked the carbohydrate-protein linkage between xylose and serine in normal heparan sulphate proteoglycan. The mean molecular weight of this heparan sulphate varied between 6300 and 9500, was not affected by treatment with alkali or Pronase and was 2-3-fold lower than that of chains released from heparan sulphate proteoglycan. After digestion with Pronase, only a minor fraction of chains contained serine, and after treatment with alkali and NaB3H4 reduction less than 5% of the chains exposed [3H]xylitol at the reducing terminals. These results suggested that (+)-catechin or metabolites of it acted as acceptors of heparan sulphate synthesis. In cultures treated wih cycloheximide, synthesis of heparan sulphate decreased to less than 5%. (+)-Catechin could restore the heparan sulphate synthesis to almost normal values. The (+)-catechin-induced heparan sulphate was secreted. Only a small fraction was incorporated into the plasma membrane or other cellular compartments. This may indicate that the protein core is essential for association of heparan sulphate with cellular compartments. 相似文献
4.
Shanping Wang Fei Liu Keai Sinn Tan Hooi‐Leng Ser Loh Teng‐Hern Tan Learn‐Han Lee Wen Tan 《Journal of cellular and molecular medicine》2020,24(1):722-736
Evidence demonstrates that M1 macrophage polarization promotes inflammatory disease. Here, we discovered that (R)‐salbutamol, a β2 receptor agonist, inhibits and reprograms the cellular metabolism of RAW264.7 macrophages. (R)‐salbutamol significantly inhibited LPS‐induced M1 macrophage polarization and downregulated expressions of typical M1 macrophage cytokines, including monocyte chemotactic protein‐1 (MCP‐1), interleukin‐1β (IL‐1β) and tumour necrosis factor α (TNF‐α). Also, (R)‐salbutamol significantly decreased the production of inducible nitric oxide synthase (iNOS), nitric oxide (NO) and reactive oxygen species (ROS), while increasing the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio. In contrast, (S)‐salbutamol increased the production of NO and ROS. Bioenergetic profiles showed that (R)‐salbutamol significantly reduced aerobic glycolysis and enhanced mitochondrial respiration. Untargeted metabolomics analysis demonstrated that (R)‐salbutamol modulated metabolic pathways, of which three metabolic pathways, namely, (a) phenylalanine metabolism, (b) the pentose phosphate pathway and (c) glycerophospholipid metabolism were the most noticeably impacted pathways. The effects of (R)‐salbutamol on M1 polarization were inhibited by a specific β2 receptor antagonist, ICI‐118551. These findings demonstrated that (R)‐salbutamol inhibits the M1 phenotype by downregulating aerobic glycolysis and glycerophospholipid metabolism, which may propose (R)‐salbutamol as the major pharmacologically active component of racemic salbutamol for the treatment of inflammatory diseases and highlight the medicinal value of (R)‐salbutamol. 相似文献
5.
Benjamin Lamp Angelika Url Kerstin Seitz Jürgen Eichhorn Christiane Riedel Leonie Janina Sinn Stanislav Indik Hemma K?glberger Till Rümenapf 《PloS one》2016,11(11)
European honey bees are highly important in crop pollination, increasing the value of global agricultural production by billions of dollars. Current knowledge about virulence and pathogenicity of Deformed wing virus (DWV), a major factor in honey bee colony mortality, is limited. With this study, we close the gap between field research and laboratory investigations by establishing a complete in vitro model for DWV pathogenesis. Infectious DWV was rescued from a molecular clone of a DWV-A genome that induces DWV symptoms such as crippled wings and discoloration. The expression of DWV proteins, production of infectious virus progeny, and DWV host cell tropism could be confirmed using newly generated anti-DWV monoclonal antibodies. The recombinant RNA fulfills Koch’s postulates circumventing the need of virus isolation and propagation of pure virus cultures. In conclusion, we describe the development and application of a reverse genetics system for the study of DWV pathogenesis. 相似文献
6.
Kirke L. Munch Erik Wapstra Scott Thomas Michelle Fisher David L. Sinn 《Ethology : formerly Zeitschrift fur Tierpsychologie》2019,125(4):203-211
Humans differ in how they perceive, assess, and measure animal behaviour. This is problematic because strong observer bias can reduce statistical power, accuracy of scientific inference, and in the worst cases, lead to spurious results. Unfortunately, reports and studies of measurement reliability in animal behaviour studies are rare. Here, we investigated two aspects of measurement reliability in working dogs: inter‐observer agreement and criterion validity (comparing novice ratings with those given by experts). Here, we extend for the first time a powerful framework used in human psychological studies to investigate three potential aspects of (dis)agreement in nonhuman animal behaviour research: (a) that some behaviours are easier to observe than others; (b) that some subjects are easier to observe than others; and (c) that observers with different levels of experience with the subject animal give the same or different ratings. We found that novice observers with the same level of experience agreed upon measures of a wide range of behaviours. We found no evidence that age of the dogs affected agreement between these same novice observers. However, when observers with different levels of experience (i.e., novices vs. a working dog expert) assessed the same dogs, agreement appeared to be strongly affected by the measurement instrument used to assess behaviour. Given that animal behaviour research often utilizes different observers with different levels of experience, our results suggest that further tests of how different observers may measure behaviour in different ways are needed across a wider variety of organisms and measurement instruments. 相似文献
7.
We investigated the application of inelastic x-ray scattering (IXS) to lipid bilayers. This technique directly measures the dynamic structure factor S(q,omega) which is the space-time Fourier transform of the electron density correlation function of the measured system. For a multiatomic system, the analysis of S(q,omega) is usually complicated. But for multiple bilayers of lipid, S(q,omega) is dominated by chain-chain correlations within individual bilayers. Thus IXS provides a unique probe for the collective dynamics of lipid chains in a bilayer that cannot be obtained by any other method. IXS of dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylcholine + cholesterol at two different concentrations were measured. S(q,omega) was analyzed by three-mode hydrodynamic equations, including a thermal diffusive mode and two propagating acoustic modes. We obtained the dispersion curves for the phonons that represent the collective in-plane excitations of lipid chains. The effect of cholesterol on chain dynamics was detected. Our analysis shows the importance of having a high instrument resolution as well as the requirement of sufficient signal-to-noise ratio to obtain meaningful results from such an IXS experiment. The requirement on signal-to-noise also applies to molecular dynamics simulations. 相似文献
8.
Middelboe M Hagström A Blackburn N Sinn B Fischer U Borch NH Pinhassi J Simu K Lorenz MG 《Microbial ecology》2001,42(3):395-406
Viral lysis of specific bacterial populations has been suggested to be an important factor for structuring marine bacterioplankton
communities. In the present study, the influence of bacteriophages on the diversity and population dynamics of four marine
bacterial phage-host systems was studied experimentally in continuous cultures and theoretically by a mathematical model.
By use of whole genome DNA hybridization toward community DNA, we analyzed the dynamics of individual bacterial host populations
in response to the addition of their specific phage in continuous cultures of mixed bacterial assemblages. In these experiments,
viral lysis had only temporary effects on the dynamics and diversity of the individual bacterial host species. Following the
initial lysis of sensitive host cells, growth of phage-resistant clones of the added bacteria resulted in a distribution of
bacterial strains in the phage-enriched culture that was similar to that in the control culture without phages after about
50-60 h incubation. Consequently, after a time frame of 5-10 generations after lysis, it was the interspecies competition
rather than viral lysis of specific bacterial strains that was the driving force in the regulation of bacterial species composition
in these experiments. The clonal diversity, on the other hand, was strongly influenced by viral activity, since the clonal
composition of the four species in the phage-enriched culture changed completely from phage-sensitive to phage-resistant clones.
The model simulation predicted that viral lysis had a strong impact on the population dynamics, the species composition, and
the clonal composition of the bacterial community over longer time scales (weeks). However, according to the model, the overall
density of bacteria in the system was not affected by phages, since resistant clones complemented the fluctuations caused
by viral lysis. Based on the model analysis, we therefore suggest that viral lysis can have a strong influence on the dynamics
of bacterial populations in planktonic marine systems. 相似文献
9.
Paschke R Kalbitz J Paetz C Luckner M Mueller T Schmoll HJ Mueller H Sorkau E Sinn E 《Journal of inorganic biochemistry》2003,94(4):335-342
This report continues our work on new compounds which consist of three functional parts--a transport fragment, a spacer and a biologically active 'drug' component. Here cholic acid functions as the transport fragment, linked via an alkyl spacer to a carboplatin analog, representing the drug (carbo-ChAPt-Fig. 1). We describe the synthesis and characterization of the series of complexes [Pt(Cyclobutane-1,1-dicarboxylato)(diamine)], [diamine=CholCOO(CH(2))(n)CH(CH(2)NH(2))(2) and THP(CH(2))(n)CH-(CH(2)NH(2))(2), n=4, 6, 8, 11]. The compounds were characterized by elemental analysis and NMR-measurements. Cytostatic activity data are given. In general, the cytostatic activity is similar to that of the parent compound and is strongly influenced by the length of the alkyl chain spacer separating the drug and transport fragments, the ones with long chain spacers being more toxic than the parent complexes. Preliminary investigations indicate the ability of the ChAPt to break resistance of tumor cells against common platinum tumor drugs, e.g. cisplatin. They are effective even on cell lines that have developed resistance to other drugs such as cis- and carboplatin. They are more cytotoxic so they are potentially effective at lower dose concentrations. The mode of cell death was examined by trypan-blue exclusion test and DNA gelelectrophoresis. Typical fragmentation of DNA was observed and the cells were still able to exclude trypan-blue. 相似文献
10.
Chen PJ Liu Y Weiss TM Huang HW Sinn H Alp EE Alatas A Said A Chen SH 《Biophysical chemistry》2003,105(2-3):721-741
We summarize a series of experimental results made with the newly developed high resolution X-ray scattering (IXS) instrument on two pure lipid bilayers, including dimyristoylphosphatidylcholine (DMPC) and dilauroylphosphatidylcholine (DLPC) in both gel and liquid crystal phases, and lipid bilayers containing cholesterol. By analyzing the IXS data based on the generalized three effective eigenmode model (GTEE), we obtain dispersion relations of the high frequency density oscillations (phonons) of lipid molecules in these bilayers. We then compare the dispersion relations of pure lipid bilayers of different chain lengths among themselves and the dispersion relations of pure lipid bilayers with those of the cholesterol containing bilayers. We also compare our experimental results with collective dynamics data generated by computer molecular dynamics (MD) simulations for dipalmitoylphosphatidylcholine (DPPC) in gel phase and DMPC in liquid crystal phase. 相似文献