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1.
Three phase partitioning (TPP) uses t-butanol and ammonium sulfate to precipitate enzymes and proteins from aqueous solutions. The method is useful both upstream with crude samples and downstream where a scaleable simple step is needed. About 25 enzymes and proteins have been isolated by various laboratories using TPP-t-butanol. The relation of t-butanol used in TPP, with n-butanol used as an extraction agent from Morton's work, is reviewed. Some t-butanol appears bound to TPP-precipitated proteins which are actually protein-t-butanol coprecipitates. They float above denser aqueous salts because bound t-butanol increases their buoyancy, similar to the behavior of many lipoproteins. On redissolving TPP-precipitated enzymes, total and specific activities usually are regained and sometimes increased. Sulfate ion-in large concentrations-likely exerts itself through its kosmotropic action as in conventional salting out. t-Butanol likewise appears to be a kosmotrope and crowding agent at room temperature or above, whereas C1 and C2 cosolvents (e.g., ethanol) do not so behave except at near or below zero temperatures. However, kosmotropy is not the entire origin of TPP, nor probably of conventional salting out. Electrostatic forces, capacity to force protein conformation tightening and protein hydration shifts, also contribute. Electrostatic forces, and the tendency for salt ions to bind and tighten protein molecule conformation, are indicated by the sharp pH dependency of both conventional salting out and TPP, around pH regions where proteins undergo conformation changes. Sulfate anion is densely-perhaps extraordinarily-hydrated, adding much to its effective size, and therefore it has a tendency to crowd or exclude proteins, when sulfate concentrations are in the 0.5 to 3 M range. 相似文献
2.
As a model for organic ferromagnetism in a one-dimensional system, substituted polyacetylenes are re-considered on the basis of the molecular orbital (MO) and the crystal orbital (CO) methods. The semiempirical MO calculations with configuration interaction for the dimer model show that the exchange interaction on poly[(4-oxyphenyl)acetylene] (1) is negative in spite of Ovchinnikov's prediction, due to the direct interaction between the adjacent pendant spins. On the other hand, it is shown that a polyacetylene chain with phenoxy radicals as pendants on every other active site can become a one-dimensional feromagnet. Moreover, the CO calculations by means of the unrestricted Hartree-Fock method give a theoretical background for the realization of a ground state with macroscopic spin alignment on the improved model chain. 相似文献
3.
We have investigated the effect of methylprednisolone on the intracellular activity of erythromycin and clindamycin in vitro. An assay system was developed for the determination of intracellular activity of antibiotics against Legionella pneumophila using guinea pig resident alveolar macrophages. Erythromycin at a concentration of 0.625 mg/L (5 x MIC) and clindamycin at a concentration of 8 mg/L (MIC) inhibited the growth of a single strain of L. pneumophila in macrophages, whilst ceftizoxime at a concentration of 0.625 mg/L (5 x MIC) did not. Methylprednisolone at therapeutic concentrations did not affect the intracellular antibacterial activity of either erythromycin or clindamycin against L. pneumophila. We found no direct effect of methylprednisolone on the intracellular antibacterial activity of either erythromycin or clindamycin. 相似文献
4.
F Higa N Kusano M Tateyama T Shinzato N Arakaki K Kawakami A Saito 《Canadian Metallurgical Quarterly》1998,36(5):1392-1398
We developed a new simple assay for the quantitation of the activities of drugs against intracellular Legionella pneumophila. The cells of a murine macrophage-like cell line (J774.1 cells) allowed the intracellular growth and replication of the bacteria, which ultimately resulted in cell death. The infected J774.1 cell monolayers in 96-well microplates were first treated with antibiotics and were further cultured for 72 h. The number of viable J774.1 cells in each well was quantified by a colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and an enzyme-linked immunosorbent assay reader. The number of growing bacteria in each well was also determined by counting the numbers of CFU on buffered charcoal yeast extract-alpha agar plates. Viable J774.1 cell counts, determined by the colorimetric assay, were inversely proportional to the number of intracellular replicating bacteria. The minimum extracellular concentrations (MIECs) of 24 antibiotics causing inhibition of intracellular growth of L. pneumophila were determined by the colorimetric assay system. The MIECs of beta-lactams and aminoglycosides were markedly higher than the MICs in buffered yeast extract-alpha broth. The MIECs of macrolides, fluoroquinolones, rifampin, and minocycline were similar to the respective MICs. According to their intracellular activities, clarithromycin and sparfloxacin were the most potent among the macrolides or fluoroquinolones tested in this study. Our results indicated that the MTT assay system allows comparative and quantitative evaluations of the intracellular activities of antibiotics and efficient processing of a large number of samples. 相似文献
5.
Eiichi Hoshino Kazunari Maruta Yasunao Wada Kazuo Mori 《Journal of the American Oil Chemists' Society》1995,72(7):785-791
The interaction of highly purified alkaline protease fromBacillus sp. KSM-K16 with the horny cells of human skin contained in skin grime was directly visualized by electron microscopy. It
became clear that the protease first penetrates the horny cells and then adsorbs, mainly onto the internal structure of the
cells at the initial stage of hydrolysis, and directly hydrolyzes the keratin filaments, though the marginal band surrounding
them retains its original shape. Then, hydrolysate produced from the keratin filaments flows out of the cell, and early in
the hydrolysis process keratin filaments decrease and then disappear, leaving a marginal band, i.e., the cell turns to a hollow
state. As a result, the remaining marginal band loses support from inside the cell, thus promoting cleavage and dispersion.
Until this stage in the protease reaction, the remarkable liberation of hydrolysis products as water-soluble protein does
not occur. 相似文献
6.
7.
Three hundred students across eight countries and eleven higher education institutions in the Asia Pacific Region participated in two courses on climate change and disaster management that were supported by learning technologies: a satellite-enabled video-conferencing system and a learning management system. Evaluation of the student experience across the region proved a significant challenge, particularly the way the students conceived of the links between the technologies and their studies, and the way they approached the use of the technologies in connected and sometimes disconnected ways. This study examines the results of a quantitative investigation into the student experience and identifies key aspects of the structure of the variables used to evaluate the experience as well as identifying groups of students in the sample population who reported qualitatively different experiences. Significant outcomes reveal that cohesive conceptions of the learning technologies tend to be related to more effective ways of using both the video-conferencing and learning management systems, and that both at the level of variables, and at the level of groups of students, these experiences tend to be related to relatively higher outcomes. The results have important implications for both the design and teaching of technology-mediated courses and offer ideas for courses that combine systems such as learning management and video-conferencing. 相似文献
8.
Stephen Matthew Lyth Huaiyu Shao Jianfeng Liu Kazunari Sasaki Etsuo Akiba 《International Journal of Hydrogen Energy》2014
Hydrogen storage is a crucial technology for the realization of a carbon-neutral society. However, few materials have been able to approach useful hydrogen storage capacity at reasonable temperatures and pressures. Graphene has an extremely high surface-area-to-weight ratio, is strong, cheap, chemically inert, and environmentally benign. As such it may be an ideal substrate for hydrogen storage. Here we present synthesis of graphene foam by combustion of sodium ethoxide. This technique is low-cost, scalable, and results in a three-dimensional graphene network with a surface area of more than 1200 m2/g. It is applied as a hydrogen storage material at liquid nitrogen temperature, with a capacity of 2.1 wt%. 相似文献
9.
10.
Ryuta Shigefuku Hideaki Takahashi Hiroyasu Nakano Tsunamasa Watanabe Kotaro Matsunaga Nobuyuki Matsumoto Masaki Kato Ryo Morita Yousuke Michikawa Tomohiro Tamura Tetsuya Hiraishi Nobuhiro Hattori Yohei Noguchi Kazunari Nakahara Hiroki Ikeda Toshiya Ishii Chiaki Okuse Shigeru Sase Fumio Itoh Michihiro Suzuki 《International journal of molecular sciences》2016,17(9)
The progression of chronic liver disease differs by etiology. The aim of this study was to elucidate the difference in disease progression between chronic hepatitis C (CHC) and nonalcoholic fatty liver disease (NAFLD) by means of fibrosis markers, liver function, and hepatic tissue blood flow (TBF). Xenon computed tomography (Xe-CT) was performed in 139 patients with NAFLD and 152 patients with CHC (including liver cirrhosis (LC)). The cutoff values for fibrosis markers were compared between NAFLD and CHC, and correlations between hepatic TBF and liver function tests were examined at each fibrosis stage. The cutoff values for detection of the advanced fibrosis stage were lower in NAFLD than in CHC. Although portal venous TBF (PVTBF) correlated with liver function tests, PVTBF in initial LC caused by nonalcoholic steatohepatitis (NASH-LC) was significantly lower than that in hepatitis C virus (C-LC) (p = 0.014). Conversely, the liver function tests in NASH-LC were higher than those in C-LC (p < 0.05). It is important to recognize the difference between NAFLD and CHC. We concluded that changes in hepatic blood flow occurred during the earliest stage of hepatic fibrosis in patients with NAFLD; therefore, patients with NAFLD need to be followed carefully. 相似文献