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41.
Hisakazu Mihara Kin-Ya Tomizaki Norikazu Nishino Tsutomu Fujimoto Haruhiko Tamaoki Yuji Kobayashi 《Biopolymers》1994,34(7):963-967
A model 16-peptide of endothelin-1 (MET-1), which has the minimized sequence homology to the corresponding pan of endothelin-1 (ET-1), was designed to confirm the cystine-stabilized α-helix motif. The model structure consists of an extended structure, a β-turn part, and an α-helix structure that is stabilized by two disulfide bonds. The α-helix segment was designed to emphasize the amphiphilic nature. In order to combine the extended structure and the α-helix segment, a D -Ala-Pro sequence was selected to fix the β-turn. The model endothelin 16-peptide amide was synthesized by solid-phase synthesis on a 4-methylbenzhydrylamine resin. Its conformation was examined by CD and two-dimensional (2D) 1H-nmr measurements. MET-1 showed similar CD patterns to ET-1 in both buffer and 50% aqueous trifluoroethanol solution. The 2D nmr experiments in 50% aqueous ethylene glycol revealed that MET-1 closely resembles the conformation of ET-1 with an extended structure, an α-helix, and a β-turn unit in the same position of the sequence. Furthermore, model peptides without disulfide bond(s) could not assume a stable structure in aqueous solution, while they did have similar α-helical content in 50% trifluoroethanol with MET-1. When the two disulfide bridges were simultaneously formed, the peptide with the correct disulfide bonds (MET-1) was obtained in threefold excess to the isomer (apamin type. MET-2). These findings obtained by the modeling of ET-1 showed an important role for the stabilization of peptide conformation with disulfide bonds. © 1994 John Wiley & Sons, Inc. 相似文献
42.
Shinichi Kitamura Takashi Hirano Kenichi Takeo Mitsuru Mimura Kanji Kajiwara Bjrn T. Stokke Tsutomu Harada 《International journal of biological macromolecules》1994,16(6)
The conformation and dilute solution properties of (2→1)-β-d-fructan in aqueous solution were studied by gel permeation chromatography, low-angle laser light-scattering photometry, viscometry, small-angle X-ray scattering and electron microscopy. Fractions covering a broad range of weight-average molecular weights (Mw) from 1.49 × 104 to 5.29 × 106 were obtained from a native sample by ultrasonic degradation and fractional precipitation. For Mw < 4 × 104, the intrinsic viscosity [η] varies with Mw0.71, indicating that the fructan chain behaves as a random coil expanded by an excluded-volume effect in this molecular weight region. For Mw > 105, [η] exhibits an unusually weak dependence on Mw and finally becomes almost independent of molecular weight. This behaviour is interpreted in terms of a globular conformation of the high-molecular-weight fructan molecules. Small-angle X-ray-scattering measurements and electron microscopic observations support this interpretation of the values of [η] observed. 相似文献
43.
A genomic clone for the cyc07 gene, which is expressed specifically at the S phase during the cell cycle in synchronous cultures of periwinkle (Catharanthus roseus) cells, was isolated. Determination of the nucleotide sequence of the clone revealed that the cyc07 gene consists of seven exons separated by six introns. Genomic Southern analysis indicated that the cyc07 gene is present as a single copy per haploid genome in periwinkle. Expression of related genes was detected in a wide range of other plants. Transgenic Arabidopsis plants were generated that expressed the gene for -glucuronidase (GUS) under the control of the promoter of the cyc07 gene. The tissue-specific pattern of expression directed by the promoter was investigated by analysis of GUS activity. Histochemical tests demonstrated that 589 bp of the 5-upstream sequence of the cyc07 gene could direct specifical expression of the GUS reporter gene in meristematic tissues in transgenic plants. The spatial pattern of expression directed by the promoter was closely correlated with meristematic activity and cell proliferation, suggesting an association between the function of the cyc07 gene and cell proliferation. 相似文献
44.
45.
A novel complex mutation in the LDL receptor gene probably caused by the simultaneous occurrence of deletion and insertion in the same region 总被引:3,自引:0,他引:3
Kimiko Yamakawa-Kobayashi Tsutomu Kobayashi Hisako Yanagi Yae Shimakura Juichi Satoh Hideo Hamaguchi 《Human genetics》1994,93(6):625-628
A novel complex mutation with the presence of both deletion and insertion in very close proximity in the same region was detected in exon 8 of the LDL receptor gene from two apparently unrelated Japanese families with familial hypercholesterolemia (FH). In this mutant LDL receptor gene, the nine bases from nucleotide (nt) 1115 to nt 1123 (AGGGTGGCT) were replaced by six different bases (CACTGA), and consequently the four amino acids from codon 351 to 354, Glu-Gly-Gly-Tyr, were replaced by three amino acids, Ala-Leu-Asn, in the conserved amino acid region of the growth factor repeat B of the LDL receptor. The nature of the amino acid substitution and data on the families suggest that this mutation is very likely to affect the LDL receptor function and cause FH. The generation of this complex mutation can be explained by the simultaneous occurrence of deletion and insertion through the formation of a hairpin-loop structure mediated by inverted repeat sequences. Thus this mutation supports the hypothesis that inverted repeat sequences influence the stability of a given gene and promote human gene mutations. 相似文献
46.
Yoshiyuki Kawakami Ichiro Ueno Tsutomu Katsuyama Ken'ichi Furihata Hideki Matsumoto 《Microbiology and immunology》1994,38(11):891-895
Epidemiological typing, based on restriction fragment length polymorphism (RFLP) by pulsed-field gel electrophoresis (PFGE), was attempted for the 38 clinical isolates of Moraxella catarrhalis obtained at Shinshu University Hospital during the years 1987 and 1993. Digestion with SmaI or NotI generated well separable, 12 to 5 genomic DNA fragments ranging from 1,000 kb to 30 kb and the strains could be classified into 14 or 13 types, respectively. The electrophoretic profile differed with the strain in most of them and was hence useful to distinguish the each strain. Investigation for their RFLP have, however, suggested that majority of them, including the type strain ATCC25238, may have derived from a common ancestor. 相似文献
47.
Takeo Uchiyama Masuro Numata Seiji Terada Tsutomu Hosino 《Plant Cell, Tissue and Organ Culture》1993,32(2):153-159
A suspension culture of Mentha was established from callus which formed on the tips of young shoots of a Mentha hybrid (M. arvenis × M. spicata). Changes in growth parameters during a culture cycle were recorded. The general appearance of cells during division and growth, including the changes in cell form, was also represented.Suspension-cultured cells of Mentha hybrid released a large amount of extracellular polysaccharides (ECP) mainly at the logarithmic phase of the growth cycle. The ECP contained galacturonic acid as major components and arabinose, galactose, glucose, xylose, rhamnose and mannose as minor components. The ratio of the uronic acid content to total sugar content in the ECP was below 40% at day 7, but increased up to 90% at day 21. The relative contents of xylose and glucose in the ECP decreased during the culture period, while the arabinose content increased and those of rhamnose, mannose and galactose remained constant.The IR spectrum suggested that the ECP were low-methoxylated pectic polysaccharides. The presence of lignin and related compounds in the ECP was not detected. The protein content of the ECP was about 10% and the main amino acids were alanine, proline, hydroxyproline, valine, asparticacid and serine, in that order. 相似文献
48.
Tsutomu Araki Hiroyuki Kato Yasuo Kanai Kyuya Kogure 《Neurochemistry international》1993,23(6):541-548
Age-related alterations in major neurotransmitter receptors and voltage dependent calcium channels were analyzed by receptor autoradiography in the gerbil brain. [3H]Quinuclidinyl benzilate (QNB). [3H]cyclohexyladenosine (CHA), [3H]muscimol, [3H]MK-801, [3H]SCH 23390, [3H]naloxone, and [3H]PN200-110 were used to label muscarinic acetylcholine receptors, adenosine A1 receptors, γ-aminobutyric acidA (GABAA) receptors,
(NMDA) receptors, dopamine D1 receptors, opioid receptors, and voltage dependent calcium channels, respectively. In middle-aged gerbils (16 months old), the hippocampus exhibited a significant elevation in [3H]QNB, [3H]MK-801, [3H]SCH 23390, [3H]naloxone, and [3H]PN200-110 binding, whereas [3H]CHA and [3H]muscimol binding showed a significant reduction in this area, compared with that of young animals (1 month). On the other hand, the cerebellum showed a significant alteration in [3H]QNB, [3H]CHA, and [3H]naloxone binding and the striatum also exhibited a significant alteration in [3H]SCH 23390 and [3H]CHA binding in middle-aged gerbils. The neocortex showed a significant elevation only in [3H]CHA binding in middle-aged animals. The nucleus accumbens and thalamus also showed a significant alteration only in [3H]muscimol binding. However, the hypothalamus and substantia nigra exhibited no significant alteration in these bindings in middle-aged gerbils. These results demonstrate the age-related alterations of various neurotransmitter receptors and voltage dependent calcium channels in most brain regions. Furthermore, they suggest that the hippocampus is most susceptible to aging processes and is altered at an early stage of senescence. 相似文献
49.
Examination of the method for measuring soil respiration in cultivated land: Effect of carbon dioxide concentration on soil respiration 总被引:7,自引:1,他引:6
Toshie Nakadai Hiroshi Koizumi Youzou Usami Mitsumasa Satoh Takehisa Oikawa 《Ecological Research》1993,8(1):65-71
An acceleration of soil respiration with decreasing CO2 concentration was suggested in the field measurements. The result supporrs that obtained in laboratory experiments in our
previous study. The CO2 concentrations in a chamber of the alkali absorption method (the AA-method) were about 150–250 parts/106 lower than that in the atmosphere (about 350 parts/106), while those observed in the open-flow IRGA method (the OF-method) were nearly equal to the soil surface CO2 levels. The AA-method at such low CO2 levels in the chamber appears to overestimate the soil respiration. Our results showed that the rates obtained by the AA-method
were about twice as large as those by the OF-method in field and laboratory measurements. This finding has important consequences
with respect to the validity of the existing data obtained by the AA-method and the estimation of changes in the terrestrial
carbon flow with elevated CO2 相似文献
50.
Isao Sakita Takushi Monden Hirohito Nagaoka Yoshihiro Katsumoto Taro Wakasugi Naohiro Tomita Tsutomu Takeda Tetsuro Kobayashi Takashi Shimano Takesada Mori 《Biotherapy》1993,6(2):103-112
We have previously reported that the antitumor effect of OK-432, a streptococcal preparation, is markedly augmented when injected intratumorally together with fibrinogen (OK-432/fbg) [1]. In order to elucidate the effects of this immunotherapy on regional lymph nodes (RLN), we carried out both morphological and functional analyses of the RLN from colonic cancer patients treated with OK-432/ fbg. Computer-aided morphometry revealed that the maximal cross-sectional areas and the broadest diameters of the RLN were significantly greater (p<0.01) in patients who had undergone local immunotherapy than in patients who had not. The component structures of RLN, such as sinus, follicle and paracortex, were all enlarged in the OK-432/fbg-treated patients, and necrosis of metastatic tumors was observed. RLN lymphocytes recovered from OK-432/fbg treated patients showed elevated reactivity to phytohemagglutinin (PHA) and the stimulation index was clearly higher than that of control patients. Flow cytometric analysis revealed a predominance of T-cells, especially CD4 subsets, and higher positivity for both CD25 and HLA-DR. Furthermore, RLN lymphocytes killed more effectively K562 and Daudi cells in the patients who had had immunotherapy. These results suggest that the effect of local immunotherapy with OK-432/fbg is not restricted to the site of injection but extends to the lymph nodes, and contributes to tumor regression through the augmentation of cellular immunity.Abbreviations RLN
regional lymph node(s)
- OK-432/fbg
OK-432/fibrinogen solution
- PHA
phytohemagglutinin
- NK
natural killer
- LAK
lymphocyte activated killer 相似文献