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Biomedical subjects

T Mine

Publications and source records attributed to T Mine.

At least 55 records · Page 3Linked to original sources
PubMed1998

Evidence for chloramphenicol/H+ antiport in Cmr (MdfA) system of Escherichia coli and properties of the antiporter.

We detected chloramphenicol/H+ antiport activity in membrane vesicles of Escherichia coli and cloned a gene for the antiporter from chromosomal DNA of E. coli. Introduction of the gene into E. coli cells conferred resistance to chloramphenicol and ethidium. A slight increase in resistance to acridine orange was also observed. Elevated chloramphenicol efflux and ethidium efflux were observed in cells harboring a plasmid carrying the gene. Addition of chloramphenicol to the assay mixture reduced the efflux of ethidium. Elevated chloramphenicol/H+ antiport activity was observed in membrane vesicles prepared from cells harboring the plasmid. The pH optimum for the activity was 6.5. We sequenced the gene and deduced the amino acid sequence of its product. A sequence homology search revealed that it was same as that of Cmr (or MdfA). Thus, it became clear that Cmr (MdfA) is the chloramphenicol(and ethidium)/H+ antiporter.

Bacterial Proteins↗
PubMed1998

Relationship between the eradication of Helicobacter pylori and the healing pattern of peptic ulcer.

We investigated the relationship between the eradication of Helicobacter pylori and the stage after the eradication of this bacterium. Eighty-six patients with H. pylori who had gastric ulcer (n=45) or duodenal ulcer (n=41) were enrolled in the study. As eradication therapy, patients received 1,500 mg amoxicillin, 400 mg clarithromycin, and 30 mg lansoprazole for 2 weeks, followed by 30 mg lansoprazole for 6 weeks in patients with gastric ulcer or for 4 weeks in those with duodenal ulcer. The ulcer stages were evaluated using the indigocarmine method at the third and sixth month after entry. The overall eradication rate of H. pylori was 85% in this study. In the gastric ulcer group, 62% of H. pylori-eradicated patients and 37.5% of H. pylori-uneradicated patients showed the S2 stage (white scar) at the sixth month. In the duodenal ulcer group, 61% of H. pylori-eradicated patients showed the S2 stage and none of the H. pylori-uneradicated patients showed the S2 stage (p < 0.05). We concluded that H. pylori eradication might promote not ulcer healing but maturity of the ulcer scar, especially in duodenal ulcer patients, and that this might also be related to the reduction of ulcer relapse.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed1998

NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli.

We found that cells of Vibrio parahaemolyticus possess an energy-dependent efflux system for norfloxacin. We cloned a gene for a putative norfloxacin efflux protein from the chromosomal DNA of V. parahaemolyticus by using an Escherichia coli mutant lacking the major multidrug efflux system AcrAB as the host and sequenced the gene (norM). Cells of E. coli transformed with a plasmid carrying the norM gene showed elevated energy-dependent efflux of norfloxacin. The transformants showed elevated resistance not only to norfloxacin and ciprofloxacin but also to the structurally unrelated compounds ethidium, kanamycin, and streptomycin. These results suggest that this is a multidrug efflux system. The hydropathy pattern of the deduced amino acid sequence of NorM suggested the presence of 12 transmembrane domains. The deduced primary structure of NorM showed 57% identity and 88% similarity with that of a hypothetical E. coli membrane protein, YdhE. No reported drug efflux protein in the sequence databases showed significant sequence similarity with NorM. Thus, NorM seems to be a novel type of multidrug efflux protein. We cloned the ydhE gene from E. coli. Cells of E. coli transformed with the cloned ydhE gene showed elevated resistance to norfloxacin, ciprofloxacin, acriflavine, and tetraphenylphosphonium ion, but not to ethidium, when MICs were measured. Thus, it seems that NorM and YdhE differ somehow in substrate specificity.

Amino Acid Sequence↗
PubMed1998

Zonal differences in effects of HGF/SF and EGF on DNA synthesis in hepatocytes under fed or starved conditions.

Zonal differences of DNA synthesis in hepatocytes induced by hepatocyte growth factor and/or scatter factor (HGF/SF) and epidermal growth factor (EGF) were investigated using male Wistar rats under fed or starved conditions. Overall, DNA synthesis was greater in fed rats than in starved rats. The predominance of EGF in periportal hepatocytes (PPH) on zonal DNA synthesis was reversed by starved conditions, but the predominance of HGF/SF on zonal DNA synthesis in perivenous hepatocytes (PVH) was not influenced by nutritional conditions. 125I-labeled EGF and 125I-labeled HGF/SF-receptor binding studies revealed no significant difference between PPH and PVH in starved or fed rats. To investigate the mechanism of the signal transduction pathway, we used genistein, an inhibitor of tyrosine kinase. Genistein had different effects on zonal difference in EGF and HGF/SF. In EGF, 1 microgram/ml genistein abolished zonal differences, but in HGF/SF 1 microgram/ml genistein did not abolish zonal differences. These data suggest that, in contrast to HGF/SF, zonal difference of DNA synthesis by EGF was dependent on nutritional conditions and DNA synthesis induced by HGF/SF and EGF might be related to tyrosine kinase, but the influence of tyrosine kinase on DNA synthesis was different between HGF/SF and EGF.

Animals↗
PubMed1997

Relationship between healing of acetic acid-induced chronic gastric ulcer and connexin.

This study examined the relationship between healing of acetic acid-induced chronic gastric ulcer and connexin formation. In addition, the effect of anti-ulcer drugs on ulcer healing and the presence of connexin was investigated. In a rat model, acetic acid-induced gastric ulcers were healed without administration of drugs after 14 days. Appearance of an electrophoretic connexin 32 band was observed 7 days after ulcer induction. Administration of cimetidine (3 mg/day) promoted ulcer healing, i.e., ulcers were healed 12 days after ulcer induction, 2 days earlier than the control. The appearance of a connexin 32 band in electrophoresis was observed on the fourth day after ulcer induction. Administration of cimetidine at a higher dose further promoted ulcer healing and the connexin 32 band was more strongly visible. After administration of i.p. 2 nM EGF, acetic acid-induced gastric ulcers were healed 12 days after ulcer induction and the appearance of connexin 32 was observed on the fourth day after induction. These results indicate that connexin 32 is related to the healing of acetic acid-induced gastric ulcers.

Acetic Acid↗
PubMed1997

[Urease-negative Helicobacter pylori isolates from gastrointestinal mucosa of patients with peptic ulcer].

It is well known that Helicobacter pylori produced a large amount of urease which plays an important role in the maintenance of infection and adhesion. Two thousand three hundred and thirty-one specimens of the gastrointestinal mucosa were obtained from patients with peptic ulcer to isolate H. pylori strains. Of these specimens, 1602 strains of H. pylori were isolated and 7 urease-negative H. pylori strains were found. Biological characteristics (except urease production) of urease-negative H. pylori were in accord with urease-positive reference strains (ATCC strains). Furthermore, these urease-negative strains were confirmed to be H. pylori strains by PCR. These results suggest that the emergence of urease-negative strains pose a new problem for H. pylori infection in patients with peptic ulcer.

Gastric Mucosa↗
PubMed1996

Betacellulin and activin A coordinately convert amylase-secreting pancreatic AR42J cells into insulin-secreting cells.

Rat pancreatic AR42J cells possess exocrine and neuroendocrine properties. Activin A induces morphological changes and converts them into neuron-like cells. In activin-treated cells, mRNA for pancreatic polypeptide (PP) but not that for either insulin or glucagon was detected by reverse transcription-PCR. About 25% of the cells were stained by anti-PP antibody. When AR42J cells were incubated with betacellulin, a small portion of the cells were stained positively with antiinsulin and anti-PP antibodies. The effect of betacellulin was dose dependent, being maximal at 2 nM. Approximately 4% of the cells became insulin positive at this concentration, and mRNAs for insulin and PP were detected. When AR42J cells were incubated with a combination of betacellulin and activin A, approximately 10% of the cells became insulin positive. Morphologically, the insulin-positive cells were composed of two types of cells: neuron-like and round-shaped cells. Immunoreactive PP was found in the latter type of cells. The mRNAs for insulin, PP, glucose transporter 2, and glucokinase, but not glucagon, were detected. Depolarizing concentration of potassium, tolbutamide, carbachol, and glucagon-like peptide-1 stimulated the release of immunoreactive insulin. These results indicate that betacellulin and activin A convert amylase-secreting AR42J cells into cells secreting insulin. AR42J cells provide a model system to study the formation of pancreatic endocrine cells.

Activins↗
PubMed1996

Norepinephrine reverses the effects of activin A on DNA synthesis and apoptosis in cultured rat hepatocytes.

Activin A, an autocrine factor produced by hepatocytes, inhibits mitogen-stimulated DNA synthesis and induces apoptotic death of cultured rat hepatocytes. Several lines of evidence indicate that norepinephrine (NE), as a comitogenic growth factor, alters the balance between growth stimulation and inhibition and acts as a trigger for the initiation of hepatocyte proliferation. In the present study, we examined whether NE modulated the effects of activin A on rat hepatocytes in primary culture. Activin A, at a concentration of 10(-9) mol/L, blocked the effect of epidermal growth factor (EGF) on DNA synthesis, that was assessed by measuring [3H] thymidine incorporation and nuclear labeling, almost completely, and NE reversed the inhibitory effect of activin A on DNA synthesis. This effect of NE was dose-dependent, being significant at concentrations of 10(-6) mol/L and above, but was overcome by higher concentrations of activin A, and was attenuated by prazosin, but not by yohimbine or propranolol. NE exerted its effect during the first 24 hours of culture, but was ineffective when added after 24 hours. EGF augmented the release of follistatin, an activin-binding protein known to block the action of activin A, by hepatocytes and NE did not affect the amount of follistatin they released. In addition to inhibiting DNA synthesis by hepatocytes cultured with EGF, activin A induced death of hepatocytes cultured in the absence of EGF. The nuclear morphology of cells cultured with activin A alone was strikingly changed compared with untreated control cells and marked identation of the nuclear membranes and moderate chromatin condensation were observed. Fragmentation of DNA was also observed, suggesting that activin A induced apoptosis, and activin-mediated cell death was prevented significantly by NE. These results indicate that NE, acting on alpha 1-adrenergic receptors, attenuates the effects of activin A on DNA synthesis by and apoptosis of cultured rat hepatocytes.

Activins↗
PubMed1996

Intravenous administration of follistatin: delivery to the liver and effect on liver regeneration after partial hepatectomy.

When 1 microgram 125I-follistatin was administered into a rat intravenously, radioactivity levels in serum decreased rapidly. Analysis with a biexponential equation showed that the initial half-life and the terminal half-life were 4.0 and 130.8 minutes, respectively. After 2 hours of infusion, approximately 9% of the follistatin infused remained in the liver, which was much more than that in kidney, spleen, pancreas, intestine, or lung. Autoradiography of the liver obtained at 24 hours of infusion revealed that numerous grains were located in parenchymal cells. Radioactivity of 125I-follistatin in the liver remained elevated until 72 hours and declined markedly thereafter. When a booster shot of 125I-follistatin was administered at 72 hours, radioactivity in the liver at 120 hours was markedly increased compared with that in rats that received a single shot of 125I-follistatin. We then examined the effect of intravenous infusion of follistatin on liver regeneration after hepatectomy of 70%. Immediately after the hepatectomy, either 1 microgram follistatin or saline was infused intravenously. In some rats, a booster shot was infused at 72 hours. After 120 hours of hepatectomy of 70%, remnant liver weight, liver regeneration rate, and DNA content were significantly (P < .05) higher in rats that received a booster shot of follistatin at 72 hours than those in control rats. These results indicate that follistatin administered intravenously accumulates in the liver and promotes liver regeneration after partial hepatectomy.

Animals↗
PubMed1996

Characterization of the activin receptor in cultured rat hepatocytes.

Activin A is an autocrine inhibitor of initiation of DNA synthesis in rat hepatocytes. The present study was conducted to characterize the cell-surface receptors for activin A in cultured rat hepatocytes by measuring 125I-activin A binding. Scatchard analysis of 125I-activin A binding indicated the existence of two classes of binding sites with apparent Kd values of 3 x 10(-10) mol/L and 3.5 x 10(-9) mol/L. Pretreatment of the cells with heparitinase reduced the number of low-affinity binding sites, whereas pretreatment with excess exogenous follistatin increased the number of low-affinity binding sites. Affinity cross-linking of 125I-activin A to hepatocytes revealed distinct protein complexes with molecular weights of approximately 48, 65, and 85 kd, which may represent cross-linked cell-bound follistatin, type I and type II activin receptors, respectively. Another band with a molecular weight of 180 kd was also found, which may represent the type III activin receptor. When hepatocytes were cultured with epidermal growth factor (EGF), both high- and low-affinity binding sites increased at 12 hours without altering their affinities. At 60 hours of the incubation with EGF, the high-affinity binding sites decreased while the number of low-affinity binding sites increased slightly. These results indicate that two classes of 125I-activin A binding sites exist in cultured hepatocytes: the high-affinity binding site may represent oligomeric complex of the type I and type II receptors, and at least part of the low-affinity binding site may represent cell-bound follistatin. The number of activin receptors in hepatocytes is increased after the stimulation with EGF.

Activin Receptors↗
PubMed1996

A detoxication route for acetaldehyde: metabolism of diacetyl, acetoin, and 2,3-butanediol in liver homogenate and perfused liver of rats.

The metabolism of diacetyl (2,3-butanedione), acetoin (3-hydroxy-2-butanone), and 2,3-butanediol, which are metabolites of acetaldehyde was quantitatively investigated using rat liver homogenate, liver perfusion, and in vivo experiments. Diacetyl and acetoin were reduced to 2,3-butanediol in these experiments, but acetoin and 2,3-butanediol were scarcely oxidized to diacetyl, indicating that the reduction reaction to 2,3-butanediol from diacetyl occurs actively in rat liver. The formation of acetoin from diacetyl required either NADH or NADPH as a reductant, while the reduction of acetoin to 2,3-butanediol required NADH. Acetoin and 2,3-butanediol were more readily accumulated than diacetyl in brain tissue.

Acetaldehyde↗
PubMed1996

Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor.

Pancreatic AR42J cells are derived from acinar cells and express both exocrine and neuroendocrine properties. We have recently shown that these cells convert into insulin-producing cells in vitro after treatment with activin A and betacellulin. Here, we investigated the effect of hepatocyte growth factor (HGF) in those cells. When AR42J cells were incubated with HGF, DNA synthesis was attenuated, and the amylase content was reduced in a concentration-dependent manner. HGF-treated cells extended processes, but bundle formation was not observed using an antibody against tubulin. Reverse both insulin and pancreatic polypeptide (PP) were expressed in HGF-treated, but not naive, AR42J cells. Immunocytochemical analysis indicated that approximately 3% of the HGF-treated cells were stained with antiinsulin antibody, and some were also stained with anti-PP antibody. When AR42J cells were exposed to a combination of activin A and HGF, cells extended longer processes, and over 10% of them were stained with antiinsulin antibody. In these cells, messenger RNAs for insulin, PP, glucose transporter 2, and glucokinase, but not those for glucagon or somatostatin, were expressed. A subclone of AR42J cells, AR42J-B13, was obtained. Most of the AR42J-B13 cells converted to insulin-producing cells after the incubation with activin A and HGF. Insulin secretion was augmented by tolbutamide, depolarizing concentrations of potassium, carbachol, and glucagon-like peptide-1 in these cells. These results indicate that HGF reduces the acinar cell-like property of AR42J cells and converts them into insulin-producing cells. The effect of HGF was markedly enhanced by activin A.

Activins↗
PubMed1995

Ultrastructural observations on the ossification of the supraspinous ligament.

STUDY DESIGN: This study analyzed the process of ossification of spinal ligaments. Supraspinous ligaments excised during surgery were studied by light and scanning electron microscopy. OBJECTIVES: The results were correlated to determine the mechanism of ossification of spinal ligaments. SUMMARY OF BACKGROUND DATA: Ossification of the ligamentum flavum has been described in detail by Hiraoka, Yamaguchi, and others. However, the pathogenesis of ossification of the spinal ligaments remains unclear. Some studies have been performed by light and transmission electron microscopy, but no detailed investigation of the ossification of the spinal ligaments by scanning electron microscopy has been performed. METHODS: Specimens of supraspinous ligament were taken from 41 patients during spinal surgery. Ossification was diagnosed macroscopically and radiologically for 20 patients. The specimens were examined using light and scanning electron microscopy. RESULTS: Collagen fibrils were 700-2000 A in diameter and were arranged in parallel. The ligament insertion divided into four zones and seemed to fit the description of Enthesis histologically. Close to the ossification region, there were a region in which some fibrils were thinner and branchings became slightly stronger, tending to form bridges of minute fibrils between other fibrils. Closer to the region of actual ossification, there was a region in which extra-fibrillar substances completely deposited. In the region of actual ossification, there were medullary spaces of varying sizes, and the surrounding collagen fibers were dense and arranged in a lamellar fashion. Osteocyte lacunae had formed and the cells regarded to be the osteocytes were present on the inside. CONCLUSION: Ossification of the supraspinous ligament possibly occurs as follows. Fibroblasts or chondrocyte-like cells respond to some external stimulus, form an irregular network of fine fibrils, and produce acid mucopolysaccharide. These undergo calcification and capillary invasion. Undifferentiated mesenchymal cells invaded and are transformed into osteoblasts. Then osteogenesis ensues with progressive calcification.

Adolescent↗
PubMed1995

A single intraportal administration of follistatin accelerates liver regeneration in partially hepatectomized rats.

BACKGROUND/AIMS: Activin A is an autocrine negative regulator of DNA synthesis in rat hepatocytes and is expressed in remnant liver after partial hepatectomy. To determine the role of activin A in liver regeneration, the effects of exogenous follistatin, which blocks the action of activin A, were examined. METHODS: Human recombinant follistatin was infused into the portal vein immediately after 70% hepatectomy. Changes in body weight, remnant liver weight, liver regeneration rate, and nuclear bromodeoxyuridine labeling were measured. RESULTS: In control rats, nuclear labeling was observed at 24 hours and peaked at 36 hours after the hepatectomy. In follistatin-treated rats, nuclear labeling was first observed after 18 hours and was significantly (P < 0.05) greater than that in control rats at 24 hours. In follistatin-treated rats, both remnant liver weight and liver regeneration rate were significantly greater at 120 hours. Serum concentrations of albumin and glucose remained reduced for up to 120 hours in control rats but recovered in follistatin-treated rats. CONCLUSIONS: A single administration of follistatin accelerates the initial round of DNA synthesis after partial hepatectomy. Activin A produced in remnant liver may exert tonic inhibitory effect on liver regeneration. Follistatin may be useful as a potential therapeutic agent to promote liver regeneration.

Activins↗
PubMed1995

Derangements in the activin-follistatin system in hepatoma cells.

BACKGROUND/AIMS: The growth of normal hepatocytes is regulated by the activin-follistatin system. The aim of this study was to investigate the activin-follistatin system in hepatoma cells. METHODS: The production and action of activin and follistatin in human hepatoma cell lines were examined. Activin A and follistatin were measured by bioassay and protein-binding assay, respectively. RESULTS: Activin A inhibited cell growth in HepG2 cells but not in either PLC/PRF/5 or HLE cells. However, the effect of activin A in HepG2 cells was attenuated at high cell density. In HepG2 cells, two classes of activin-binding sites were expressed, and affinity cross-linking showed that 125I-activin A bound specifically to three proteins with molecular weights of 48, 67, and 94 kilodaltons. In PLC/PRF/5 cells, a single class of binding site was observed, and the binding capacity was approximately 60% of the capacity in HepG2 cells. Virtually no 125I-activin A binding was detected in HLE cells. Bioactivity and messenger RNA for activin A were undetectable in three cell lines. In contrast, follistatin was released from three cell lines. CONCLUSIONS: Multiple alterations in the activin-follistatin system were found in three hepatoma cell lines. The accelerated growth observed in hepatoma cells may be caused, at least partly, by the attenuation of the action of activin A.

Activins↗