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41.
N. Ohmiya Shinsuke Saga Masaharu Ohbayashi Ken-ichi Kozaki Osamu Miyaishi Miya Kobayashi Shiro Kasuya Tomiyasu Arisawa Hidemi Goto Tetsuo Hayakawa 《Histochemistry and cell biology》1997,108(1):27-34
An association between eosinophils and tissue damage has been observed in numerous disorders. However, few reports have addressed
the role of infiltrating eosinophils in gastric ulcer healing. The aim of this study was to investigate the kinetics and role
of eosinophils infiltrating experimental chronic gastric ulcers in the rat. We developed a monoclonal antibody against human
matrix metalloproteinase 1 (MMP1) purified from conditioned culture medium of human skin fibroblasts. Acetic acid-induced
gastric ulcers were resected from rats on days 1, 3, 5, 10, 20, 40, and 180 after the days of induction (day 0). Tissue specimens
were immunostained with this antibody and examined with an electron microscope. Few eosinophils were observed in the granulation
tissue until day 20. By days 40 and 180, MMP1-positive eosinophils had increased in the granulation tissue of open ulcers.
Azan staining revealed dispersed collagen fibers around infiltrating eosinophils. In contrast, scars demonstrated few eosinophils
in fibrous tissue on days 40 and 180. Eosinophils which express MMP1 infiltrate granulation tissue at the chronic stage of
gastric ulceration. The results suggest that eosinophils may play a role in tissue remodeling and deterioration of ulceration.
Accepted: 18 March 1997 相似文献
42.
Ayako Yamamoto Tetsuo Hashimoto Emiko Asaga Masami Hasegawa Nobuichi Goto 《Journal of molecular evolution》1997,44(1):98-105
Major parts of amino-acid-coding regions of elongation factor (EF)-1α and EF-2 in Trichomonas tenax were amplified by PCR from total genomic DNA and the products were cloned into a plasmid vector, pGEM-T. The three clones
from each of the products of the EF-1α and EF-2 were isolated and sequenced. The insert DNAs of the clones containing EF-1α
coding regions were each 1,185 bp long with the same nucleotide sequence and contained 53.1% of G + C nucleotides. Those of
the clones containing EF-2 coding regions had two different sequences; one was 2,283 bp long and the other was 2,286 bp long,
and their G + C contents were 52.5 and 52.9%, respectively. The copy numbers of the EF-1α and EF-2 gene per chromosome were
estimated as four and two, respectively.
The deduced amino acid sequences obtained by the conceptual translation were 395 residues from EF-1α and 761 and 762 residues
from the EF-2s. The sequences were aligned with the other eukaryotic and archaebacterial EF-1αs and EF-2s, respectively.
The phylogenetic position of T. tenax was inferred by the maximum likelihood (ML) method using the EF-1α and EF-2 data sets. The EF-1α analysis suggested that
three mitochondrion-lacking protozoa, Glugea plecoglossi, Giardia lamblia, and T. tenax, respectively, diverge in this order in the very early phase of eukaryotic evolution. The EF-2 analysis also supported the
divergence of T. tenax to be immediately next to G. lamblia.
Received: 15 February 1996 / Accepted: 28 June 1996 相似文献
43.
44.
Summary The parameters for HN chemical shift calculations of proteins have been determined using data from high-resolution crystal structures of 15 proteins. Employing these chemical shift calculations for HN protons, the observed secondary structure chemical shift trends of HN protons, i.e., upfield shifts on helix formation and downfield shifts on -sheet formation, are discussed. Our calculations suggest that the main reason for the difference in NH chemical shifts in helices and sheets is not an effect from the directly hydrogen-bonded carbonyl, which gives rise to downfield shifts in both cases, but arises from an additional upfield shift predicted in helices and originating in residues i-2 and i-3. The calculations also explain the well-known relationship between amide proton shifts and hydrogen-bond lengths. In addition, the HN chemical shifts of the distorted amphipathic helices of the GCN4 leucine zipper are calculated and used to characterise the solution structure of the helices. By comparing the calculated and experimental shifts, it is shown that in general the agreement is good between residues 15 and 28. The most interesting observation is that in the N-terminal half of the zipper, although both calculated and experimental shifts show clear periodicity, they are no longer in phase. This suggests that for the N-terminal half, in the true average solution structure the period of the helix coil is longer by roughly one residue compared to the NMR structures. 相似文献
45.
H Nomura A Kitamura Y Tanigawa M Tsuchiya M Ueki O Sugimoto M Shimoyama 《Biochimica et biophysica acta》1984,781(1-2):112-120
We examined the fragmentation of DNA treated with N-methyl-N-nitrosourea under conditions in which Ca2+, Mg2+-dependent endonuclease is active. The molecular mass of DNA found in mouse liver slices treated with methylnitrosurea in the presence of Ca2+ plus Mg2+ was 4 X 10(5) Da. Similar results were obtained with a reconstituted system containing partially purified Ca2+, Mg2+-dependent endonuclease and methylnitrosurea-treated DNA. The enzyme extensively cleaved methylnitrosurea-treated DNA, compared with non-treated DNA. The methylnitrosurea-treated nuclear proteins obtained from mouse liver nuclei had no effect on the DNA fragmentation by the enzyme. Using closed-circular DNA treated with methylnitrosurea, the enzyme produced single-strand cuts in the DNA, as was seen in non-treated, closed-circular DNA, however, the rate of hydrolysis was increased. Ca2+, Mg2+-dependent endonuclease thus warrants further investigation, with regard to the precise mechanism of extensive degradation of DNA in cells treated with carcinogenic alkylating agents. 相似文献
46.
We have previously shown that a coenzyme-B12 analog, adenosylcobalamin (AdoCbl)-(e-OH), with the e-propionamide group converted to a carboxylic acid, serves as a poor coenzyme for dioldehydrase. During the course of the catalytic process, the enzyme AdoCbl-(e-OH) complex becomes catalytically inactive (T. Toraya, E. Krodel, A. S. Mildvan, and R. H. Abeles, 1979, Biochemistry18, 417–426). We have now examined the mechanism of this inactivation further. Inactivation only occurs in the presence of substrate. The dioldehydrase coenzyme analog complex is stable in the absence of substrate. In the inactivated complex, the coenzyme analog was stoichiometrically converted to a cob(II)alamin species. The cob-(II)alamin formed remained irreversibly bound at the active site of the enzyme and resisted oxidation by O2 even in the presence of CN?. Stoichiometric formation of 5′-deoxyadenosine from the 5′-deoxy-5′-adenosyl moiety of the coenzyme analog was demonstrated with [8-14C]-AdoCbl(e-OH). This nucleoside also remained tightly bound to the enzyme and was not exchangeable with free 5′-deoxyadenosine nor was it removed by Sephadex chromatography. The rate of inactivation showed no deuterium isotope effect when the inactivation occurred in the presence of l,2-propanediol-l-d2. The inactivated complex was resolved by acid ammonium sulfate treatment into the intact apoenzyme and the hydroxocobalamin derivative. This indicates that the apoenzyme itself is not modified in the inactivation process. These results suggest that the inactivation reaction occurs from one of the intermediates in the normal catalysis. We propose that the inactivation is due to incorrect binding of the modified coenzyme in an intermediate of the catalytic process. This incorrect binding leads to the loss of the substrate radical, and consequently, to loss of catalytic activity. 相似文献
47.
Sachiko Kuno Tetsuo Toraya Saburo Fukui 《Archives of biochemistry and biophysics》1980,205(1):240-245
The apoenzyme of diol dehydrase was inactivated by two arginine-specific reagents, 2,3-butanedione and phenylglyoxal, in borate buffer. In both cases, the inactivation followed pseudo-first-order kinetics. Kinetic data show that the incorporation of a single reagent molecule per active site of the enzyme is necessary for the complete inactivation. The modification with 2,3-butanedione was reversed by dilution of the reagent and borate concentrations (65% activity recovered). 1,2-Propanediol (substrate) partially protected the enzyme against inactivation. The holoenzyme was almost insensitive to 2,3-butanedione and phenylglyoxal, indicating that the essential arginine residue is prevented from the attack of these reagents either by direct blockage with the bound coenzyme or by an indirect conformational change caused by coenzyme binding. The inactivation of diol dehydrase by 2,3-butanedione did not result in dissociation of the enzyme into subunits. From these results, we concluded that the essential arginine residue is located at or in close proximity to the active site of diol dehydrase. 相似文献
48.
Upon addition of divalent cations to the incubation medium ofcultured tobacco cells, the release of phosphatase into themedium increased and the time course of the release became biphasic.A rapid release (phase I release) occurred instantaneously afterthe addition and then a release at a constant rate (phase IIrelease) followed. Sodium and potassium ions did not affectthe enzyme release. Lanthanum ions caused the biphasic enzymerelease but inhibited the phase II release. The effects of temperature and metabolic inhibitors indicatedthat phase I release was limited by a diffusion process butphase II release was limited by an enzymatic reaction requiringmetabolic energy. From the results it was concluded that divalent cations enhancedthe enzyme release not only by stimulating the transport ofenzyme to the outside of the cell membrane, but also by liberatingthe enzyme retained on the exterior of the cells, e.g., thecell walls. The released phosphatase could be separated into two fractions,F-I and F-II. Only F-I was released by phase I release, whileboth F-I and F-II resulted from phase II release. This indicatedthat F-I was preferentially trapped on the exterior of the cells.
1 These experiments were carried out at the Department of Botanyin the Faculty of Science of the University of Tokyo. (Received December 15, 1978; ) 相似文献
49.
Tetsuo Omata Noritada Iwamoto Tomio Kimura Atsuo Tanaka Saburo Fukui 《Applied microbiology and biotechnology》1981,11(4):199-204
Summary
dl-Menthyl succinate was successfully hydrolyzed stereoselectively by Rhodotorula minuta var. texensis cells entrapped within photo-crosslinked or polyurethane resin gels in water-saturated n-heptane. The hydrolyzed product was found to be pure l-menthol. The catalytic activity of the immobilized cells, especially those entrapped in urethane polymers, was far more stable than that of the free cells. The half-life of the polyurethaneentrapped cells was estimated to be 55–63 days in the organic solvent.Dedicated to the 65th birthday od Professor Dr. G. Manecke 相似文献
50.
Chromatographic separation, of extracellular acid phosphatase of tobacco cells cultured under Pi-supplied and omitted conditions 总被引:1,自引:0,他引:1
An extracellular acid phosphatase preparation of tobacco XD-6cells cultured in suspension was resolved into three fractionsby sequential chromatography. Two of these were neutral pyrophosphatasewith diesterase activity, having optimum pH at 6.8. The otheris a nonspecific acid phosphatase having optimum pH at 5.8.The latter was concluded to be involved in the increase in extracellularactivity upon Pi-depletion. (Received August 31, 1976; ) 相似文献