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T Shigematsu

Publications and source records attributed to T Shigematsu.

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

Nitric oxide modulates T-lymphocyte migration in Peyer's patches and villous submucosa of rat small intestine.

BACKGROUND & AIMS: Although nitric oxide (NO) is known to influence the recruitment of neutrophils in inflamed tissue, its role in lymphocyte-endothelial cell interactions remains poorly understood. The objectives of this study were to assess the effects of NO synthesis inhibition on T-lymphocyte migration in microvessels of rat small intestine and to define the role of adhesion molecules in this process. METHODS: T lymphocytes collected from rat intestinal lymph were labeled with carboxyfluorescein diacetate succinimidyl ester and injected into the jugular vein of recipient rats. The migration of T lymphocytes into normal and NG-nitro-L-arginine methyl ester (L-NAME)-treated intestinal microvessels was monitored by using an intravital microscope. RESULTS: L-NAME significantly increased rolling and adherence of lymphocytes in postcapillary venules of Peyer's patches and submucosal venules without significantly decreasing red blood cell velocity. The subsequent appearance of lymphocytes in the initial lymphatics was also accelerated by L-NAME. Anti-4-integrin antibody markedly inhibited the L-NAME-induced lymphocyte-endothelial cell interaction. Anti-P-selectin monoclonal antibody also significantly attenuated these adhesive interactions in both vascular regions. CONCLUSIONS: These data suggest that NO is an important modulator of lymphocyte migration in Peyer's patches and in nonlymphoid regions of the intestine.

Animals↗
PubMed1998

Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.

BACKGROUND & AIMS: The basic mechanisms of food allergies are still unknown. The aims of this study were to investigate whether endothelins (ETs) in the intestinal mucosa are involved in the pathogenesis of intestinal anaphylaxis. METHODS: Sprague-Dawley rats were sensitized to chicken egg albumin (EA) by intraperitoneal injection. Fourteen days after sensitization, EA was administered in the jejunal segments to induce intestinal anaphylaxis. Net water outflux and histamine release into loops and serum concentrations of rat mast cell protease II (RMCP-II) were determined. ET-1 and ET-3 concentrations in the jejunal mucosa were determined, and expression of the corresponding messenger RNAs was examined by competitive polymerase chain reaction. RESULTS: In sensitized animals, challenge with intraluminal antigen caused a significant increase in net water outflux and histamine release together with an elevation of serum RMCP-II concentrations. Mucosal concentrations of ET-1 and ET-3 and expression of their messenger RNAs were significantly increased in sensitized animals after EA challenge. Treatment with an ETA-receptor antagonist, but not an ETB-receptor antagonist, attenuated the increase in net water outflux, histamine release, and serum RMCP-II concentrations in rats with EA-induced intestinal anaphylaxis. CONCLUSIONS: Release of ETs in the intestinal mucosa increased in sensitized animals after EA challenge. ETs may play a significant role in the development of intestinal anaphylaxis via an ETA receptor.

Albumins↗
PubMed1998

Medical baseline data collection on bone and muscle change with space flight.

It has been documented that astronauts suffer from a progressive and continuous negative calcium balance in space flight. The National Space Development Agency of Japan (NASDA) discussed the experimental protocols with the National Aeronautics and Space Agency's (NASA's) Johnson Space Center (JSC) and has started a medical baseline collection on bone and calcium metabolism, and muscle changes with space flight. The subjects were two astronauts, a 42-year-old female and a 32-year-old male, who experienced real space flights. Fractional excretion of calcium (FECa) increased in both subjects just after the space flight. There was a negative calcium balance with urinary calcium leak even after a short flight. We also noticed a decrease (-3.0%) of bone mineral density (BMD) of the lumbar spine (L2-4), a weight bearing bone. These bone changes may be due to a negative calcium balance. However, the BMD of the skull, a nonweight bearing bone, increased after the flight. This indicates that the effect of weightlessness on bone is different in respective bones, depending on the weight loading. Our data of the bone metabolic marker clearly indicate that bone resorption is stimulated, shown by an elevation of urinary pyridinolinks and plasma tartrate-resistant acid phosphate (TRACP) activity. Bone specific alkaline phosphatase, a bone formation marker, was elevated in both subjects, but not intact osteocalcin. Whether this pathophysiological phenomenon is due to an accelerated bone resorption or suppressed bone formation is still obscure. In addition, the physiological cross-sectional area (PCSA) of muscle in the legs greatly decreased (from - 10% to -15%) after the flight, and it took over a month to be recovered in both subjects. However, the muscle volume loss in the legs seemed to be reversible. To examine bone and muscle metabolism with space flight, further investigations and international standardization of experimental protocols are necessary.

Adult↗
PubMed1998

Apa I polymorphism in the vitamin D receptor gene may affect the parathyroid response in Japanese with end-stage renal disease.

Vitamin D [1,25(OH)2D3] plays a key role in the pathogenesis of secondary hyperparathyroidism. A polymorphism in the vitamin D receptor (VDR) gene is reported to be involved in bone mineral density and the serum level of intact-osteocalcin (i-OC) in patients with osteoporosis. We investigated the relationship between VDR gene polymorphisms and the levels of intact PTH (i-PTH) and i-OC in 129 Japanese patients with end-stage renal disease (ESRD). The VDR gene sequences were PCR-amplified, and the product was cleaved with the restriction enzymes Bsm I and Apa I. Undigested alleles were designated as B and A, and the digested alleles as b and a, respectively. The frequencies for the Bsm I polymorphism were 0.0% BB, 19.4% Bb, and 80.6% bb, while those for Apa I polymorphism were 14.2% AA, 47.2% Aa, and 38.6% aa. The Bsm I polymorphism of VDR was greatly biased in Japanese people. The i-PTH level in the aa group was about twice as high as those in the both AA group and Aa group (P < or = 0.04). The i-OC concentrations in the aa group was also approximately double those in both the AA group and Aa group (P < or = 0.03). In contrast, no significant differences in age, duration of dialysis, male/female ratio, or the incidence of diabetic nephropathy were observed among these three groups. On the other hand, there was no significant differences in i-PTH and i-OC between the Bb and bb groups. These results suggest that VDR gene polymorphisms can affect parathyroid response in ESRD patients, and the Apa I polymorphism is more informative in Japanese patients than the Bsm I polymorphism. The VDR a gene allele may define the pathogenesis of secondary hyperparathyroidism and of high turnover bone disease in patients with ESRD.

Alleles↗
PubMed1998

Oxidative stress and mitochondrial damage precedes gastric mucosal cell death induced by ethanol administration.

Although it has been speculated that active oxidants and mitochondrial membrane damages play roles in ethanol-induced gastric mucosal damages, its detail remains unknown. The present study was designed to investigate whether ethanol induces oxidative stress and mitochondrial permeability transition (MPT) before cell death of gastric mucosal cells. Rat gastric mucosal cells (RGM-1) were kept in serum-free Dulbecco's modified Eagle's medium before addition of various concentrations of ethanol. Nuclear morphological aftemations and membrane barrier dysfunction of RGM-1 cells were assessed by staining with Hoechst 33342 and propidium iodide, respectively. To assess the contribution of oxygen-derived free radicals and intracellular glutathione, scavenger of hydrogen peroxide and the hydroxyl radical, N,N-dimethylthiourea, glutathione precursor, N-acetyl-L-cysteine, and an inhibitor of alcohol dehydrogenase, 4-methylpyrazole were added before treatment with ethanol. To investigate MPT, calcein and tetramethylrhodamine methyl ester were loaded before addition of ethanol, and the changes of fluorescence intensity were monitored using a laser scanning confocal microscope. Ethanol (>5% v/v) dose-dependently increased the number of propidium iodide-positive cells, suggesting a diminished barrier function of cell membrane. After addition of ethanol, mitochondria were filled quickly with calcein indicating MPT, which was accompanied by mitochondrial depolarization, as shown by loss of tetramethylrodamine methyl ester before cell death. Ethanol-induced cell death was significantly attenuated by simultaneous incubation with either N,N-dimethylthiourea or N-acetyl-L-cysteine, suggesting the importance of intracellular redox states in inducing cellular damage, whereas such change was not attenuated by 4-methylpyrazole. Present results suggest that ethanol treatment induces intracellular oxidative stress and produces MPT and mitochondrial depolarization, which are preceding cell death in gastric mucosal cells. Intracellular antioxidants, such as glutathione, may have a significant protective action against ethanol in gastric mucosal cells.

Animals↗
PubMed1998

Increased expression of an inducible isoform of nitric oxide synthase and the formation of peroxynitrite in colonic mucosa of patients with active ulcerative colitis.

BACKGROUND: Increased production of reactive metabolites of oxygen and nitrogen has been implicated in chronic inflammation of the gut. The object of this study was to examine the magnitude and location of nitric oxide synthase (NOS) activity and peroxynitrite formation in the colonic mucosa of patients with ulcerative colitis in relation to the degree of inflammation. SUBJECTS: Thirty three patients with active ulcerative colitis (17 with mild or moderate inflammation, 16 with severe inflammation). METHODS: Inducible NOS activity was determined in the colonic mucosa by measuring the conversion of L-arginine to citrulline in the absence of calcium. The localisation of NOS and nitrotyrosine immunoreactivity was assessed immunohistochemically using the labelled streptavidin biotin method. RESULTS: Inducible NOS activity increased in parallel with the degree of inflammation of the mucosa. Expression of inducible NOS was found not only in the lamina propria, but also in the surface of the epithelium. Peroxynitrite formation as assessed by nitrotyrosine staining was frequently observed in the lamina propria of actively inflamed mucosa. CONCLUSIONS: Nitric oxide and peroxynitrite formation may play an important role in causing irreversible cellular injury to the colonic mucosa in patients with active ulcerative colitis.

Acute Disease↗
PubMed1998

Induction of endothelin-1 synthesis by IL-2 and its modulation of rat intestinal epithelial cell growth.

Endothelin (ET), a vasoconstrictive peptide, is known to have a variety of biological actions. Although ET is released by vascular endothelial cells, other cell populations also have been reported to synthesize and release ET. In this study, we examined whether ET is synthesized by intestinal epithelial cells and whether it affects induction of epithelial cell proliferation by interleukin-2 (IL-2). Subconfluent monolayers of intestinal epithelial cells (IEC-6 and IEC-18) were maintained in serum-free medium before addition of rat IL-2. Both IEC-6 and IEC-18 cells released ET-1 into the medium under unstimulated conditions, as determined by a sandwich ELISA. IL-2 significantly enhanced ET-1 release in a time-dependent manner. ET-3 was not detectable in the culture media of either cell line. Expression of ET-1 and ET-3 mRNA in epithelial cells was assessed by competitive PCR. Both cell lines were shown to express ET-1 mRNA, but no ET-3 mRNA was detected. IL-2 treatment enhanced ET-1 mRNA expression by both IEC-6 and IEC-18 cells. Both cell lines also expressed mRNA for ETA and ETB receptor subtypes. When cell proliferation was assessed, exogenous ET-1 induced a slight proliferative response in both types of cells that was consistent and significant at low ET-1 concentrations; cell growth was inhibited at a higher concentration (10(-7) M). IL-2 produced a significant proliferative response in both cell lines. However, the addition of ET-1 (10(-7) M) to culture media attenuated the IL-2-induced increase in cell proliferation. ETA-receptor antagonists significantly enhanced cellular proliferation, suggesting involvement of the ETA receptor in modulation of IL-2-induced intestinal epithelial cell growth.

Animals↗
PubMed1997

Dissection of the promoter/operator region and evaluation of N-acylhomoserine lactone mediated transcriptional regulation of elastase expression in Pseudomonas aeruginosa.

In Pseudomonas aeruginosa, expression of the lasB gene which codes for the metalloprotease, elastase, depends on small diffusible N-acylhomoserine lactones. lasB expression is regulated through the interactions of N-3-oxododecanoyl-L-homoserine lactone and N-butanoyl-L-homoserine lactone with the transcriptional activators LasR and VsmR(RhlR), respectively. To investigate lasB expression further, we first located the transcriptional start site to a position 141 bp upstream from the translational start site. Using this information, we constructed a series of plasmids containing consecutive 5' deletions of the upstream region of lasB fused to a promoterless chloramphenicol acetyltransferase reporter gene. The results obtained indicate that three regions are required for efficient transcription of lasB; a 35 bp palindromic sequence located at +26 to +60 bp upstream from the translation start site, and two regions located upstream of the transcription start site, at -135 to -85 bp and -63 to -26 bp, respectively. Deletion of the latter region results in the loss of both N-butanoyl-L-homoserine lactone- and N-3-oxododecanoyl-L-homoserine lactone-mediated stimulation of lasB expression and provides further support for the role of this operator site as a target for either or both LasR and VsmR.

Bacterial Proteins↗
PubMed1997

Adenocarcinoma of the uterine cervix with predominantly villogladular papillary growth pattern.

We reviewed the clinical and pathological features of seven cases of adenocarcinoma of the uterine cervix with predominantly villogladular papillary growth pattern. The patients, who ranged in age from 33 to 54 (mean, 45) years, underwent radical hysterectomy. In all seven cases, the tumors were papillary exophytic architecture lined by stratified epithelial cells with mild to moderate nuclear atypicality. In one of seven cases, the majority of the tumor showed villogladular papillary component, but the small foci of small cell carcinoma was present in the endocervical end of the tumor. The lymph vascular invasion was demonstrated in two of seven cases, and these two had pelvic lymph node metastases. One of these two patients had recurrence 30 months after the initial treatment and died of disease after 46 months. The follow-up ranged from 9 to 169 (median, 46) months. The presence or absence of lymph vascular invasion and minor components of this tumor such as small cell carcinoma, serous cell carcinoma, and clear cell carcinoma with a poor prognosis may be important histological findings before deciding to manage this tumor by the conservative treatment.

Adenocarcinoma↗
PubMed1997

Early adenocarcinoma of the uterine cervix.

The purpose of this study was to evaluate the prognostic significance of the tumor depth, horizontal spread, and volume in early cervical adenocarcinoma while excluding adenocarcinoma in situ. Thirty cases who had been treated at our institution having cervical adenocarcinoma with a tumor depth of less than 5 mm were clinicopathologically reviewed. The volumes were estimated based on the portion with the largest tumor surface area by multiplying three dimensions: depth, horizontal spread, and a third dimension. The third dimension was calculated by the method of Burghardt to be 1.5 times the largest measured depth or spread. Two of the 30 patients recurred in the vagina at 18 and 163 months after the initial operation; the former patient died of disease 87 months postoperatively. The remaining 28 patients are all doing well without recurrence (range of follow-up from 24 to 232 months; median 79 months). No pelvic or paraaortic lymph node metastases were seen in 25 and 22 cases, respectively. None of the 21 cases with a lesion measuring less than 3 mm in depth had recurrence. On the other hand, 1 of 23 with a tumor volume up to 500 mm3 had recurrence. The estimated 5-year progression-free survival rates for patients with cervical adenocarcinoma with a depth of less than 3 mm and those with a depth of more than 3 mm were 100 and 88.89%, respectively (P = 0.116). The depth of stromal invasion may therefore be a good predictor of lymph node metastasis and recurrence in early cervical adenocarcinoma.

Adenocarcinoma↗
PubMed1997

Pressure stimulates proliferation and DNA synthesis in rat intestinal epithelial cells.

Effects of intraluminal pressure on cultured intestinal epithelial cells were assessed by measuring cell proliferation and DNA synthesis after exposure to various pressures. Pressures of 40 to 120 mm Hg promoted cell proliferation and DNA synthesis. Pressure-induced DNA synthesis was further enhanced by the addition of interleukin-2, suggesting the regulation of intestinal epithelial growth by pressure in coordination with cytokines. Pretreatment with either a phospholipase C inhibitor or protein kinase C inhibitor significantly inhibited DNA synthesis promoted by pressure and interleukin-2. This study demonstrates a novel mechanism whereby pressure regulates cell growth in intestinal epithelial cells, possibly via activation of phospholipase C and protein kinase C.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1997

Increased nitric oxide production and inducible nitric oxide synthase activity in colonic mucosa of patients with active ulcerative colitis and Crohn's disease.

It is postulated that an enhanced production of nitric oxide by inflamed intestine plays a role in the pathophysiology of active inflammatory bowel disease. In this study, systemic NOx concentrations and colonic nitric oxide synthase activity were determined in patients with ulcerative colitis or Crohn's disease. The relationship between these two parameters and disease activity, as well as differences in nitric oxide synthase activity between ulcerative colitis and Crohn's disease, were areas of specific focus. Patients with active ulcerative colitis and Crohn's disease had significantly elevated plasma NOx concentrations; a positive correlation was found between NOx values and inducible nitric oxide synthase activities in the active mucosa of these patients. In active ulcerative colitis, levels of inducible nitric oxide synthase were significantly elevated in both normal and inflamed mucosa, although inducible nitric oxide synthase activity was higher in the latter. These colonic inducible nitric oxide synthase activities correlated well with the results of endoscopic and histologic grading of inflammation. There was no increase in constitutive nitric oxide synthase activity in patients with active ulcerative colitis. However, constitutive nitric oxide synthase activity was significantly increased in the inflamed mucosa in patients with Crohn's disease. In Crohn's disease, elevated inducible nitric oxide synthase activity was found in both normal and inflamed mucosa, with no significant difference between the tissues. Such differences in nitric oxide production in the colonic mucosa possibly reflect the significant differences in the pathophysiology and characteristic clinical features between ulcerative colitis and Crohn's disease.

Adult↗
PubMed1997

Immunohistochemical detection of advanced glycosylation end-products in the peritoneum and its possible pathophysiological role in CAPD.

It has recently been suggested that advanced glycosylation end-products (AGEs) are formed in the peritoneum in patients on CAPD. However, the exact location of AGE accumulation, the relation with the duration of CAPD and its pathophysiological role in CAPD remain unclear. If the peritoneum is glycosylated, it could bring about altered peritoneal function. Therefore, the aim of this study is to clarify the localization of AGEs in the peritoneum in accordance with the duration of CAPD and to examine its relation to the peritoneal permeability. Fifteen non-diabetic patients were divided into three groups (each 5 patients) on the basis of the mean duration (D) of CAPD (Group 1, D = 0 month; Group II, D = 34 months; Group III, D = 84 months). The AGE staining by monoclonal anti-AGE antibody in the peritoneum and the four-hour peritoneal equilibration test (PET) were compared among these groups. AGE was absent or found only weakly in Group I. However, in groups II and III, AGE was moderately or strongly positive especially in the vascular walls and it was dominant in group III. PET revealed that peritoneal permeability for glucose, creatinine, beta2-microglobulin and albumin was increased in Group II as compared to Group I, and it was further increased in Group III. The results of this study indicate that AGEs become dominantly accumulated in the vascular wall in accordance with the prolongation of CAPD treatment, and this might play some roles for the increased permeability of the peritoneal membrane in CAPD.

Adult↗
PubMed1997

Enhanced lymphocyte interaction in postcapillary venules of Peyer's patches during fat absorption in rats.

BACKGROUND & AIMS: Although dietary fat is known to modulate immune functions, there is a paucity of data concerning the effects of fat absorption on migration of lymphocytes in intestinal lymphoid tissue. The objective of this study was to assess the influence of fat absorption on T-lymphocyte migration in Peyer's patches. METHODS: T lymphocytes collected from rat intestinal lymph were labeled with carboxyfluorescein diacetate succinimidyl ester and injected into the jugular vein of recipient rats. Olive oil or octanoic acid was placed into the duodenum, and Peyer's patch microcirculation was observed by intravital fluorescence microscopy. RESULTS: Rolling and adherence of lymphocytes in postcapillary venules were noted in both fasted and fed rats. However, lymphocyte adherence was significantly greater in olive oil-fed rats than in fasted rats. Olive oil also significantly increased the transendothelial migration of T lymphocytes. Octanoic acid stimulated lymphocyte rolling but did not affect lymphocyte adherence. Olive oil enhanced the expression of both alpha4-integrin and L-selectin on lymphocytes, whereas octanoic acid only increased the expression of L-selectin. CONCLUSIONS: Lymphocyte rolling and adherence are enhanced by olive oil feeding, possibly through an interaction between activated adhesion molecules on lymphocytes and counter-receptors on endothelial cells.

Animals↗
PubMed1997

Vasoactive intestinal peptide modulates T lymphocyte migration in Peyer's patches of rat small intestine.

Although vasoactive intestinal peptide (VIP) has been postulated to function in modulation of T cell trafficking, the exact mechanism has not been elucidated in vivo. In the present study, the effects of VIP on T lymphocyte migration were examined in rat Peyer's patches. T lymphocytes collected from intestinal lymph of rats were labeled with carboxyfluorescein diacetate succinimidyl ester and injected into the jugular vein. Peyer's patches of the recipient rats were observed with intravital fluorescence microscopy. In vivo intra-arterial infusion of or in vitro incubation with VIP did not affect the initial lymphocyte interaction with postcapillary venules of Peyer's patches. However, these treatments with VIP significantly inhibited transendothelial migration and also significantly blocked the interstitial migration of T cells and inhibited their subsequent appearance in the interfollicular lymphatics. Treatment with adenosine 3',5'-cyclic monophosphate (cAMP)-inducing agents resulted in similar inhibitory effect on T lymphocyte migration in Peyer's patches. In conclusion, VIP has significant inhibitory effects on T lymphocyte migration in Peyer's patches, possibly mediated by elevation of the intracellular cAMP concentrations.

Animals↗
PubMed1997

Oxygen sensitivity of NifA protein of Azospirillum lipoferum FS as suggested by gene cloning and expression in Escherichia coli.

We cloned and sequenced a 2.8-kb SalI fragment of Azospirillum lipoferum FS as a homologue of the Klebsiella oxytoca nifA gene. The amino acid sequence deduced from an open reading frame of 1872 bases showed 91% identity to that of the A. brasilense NifA, and the putative central sigma54 interaction domain was conserved as well as the C-terminal DNA-binding domain. The NifA function on the nifH promoter was examined in Escherichia coli using a combination of a nifA driver plasmid and a nifH-lacZ reporter plasmid, in which the transcriptional activation of the nifH promoter by the NifA was evaluated with the beta-galactosidase activity. The A. lipoferum NifA activated the nifH promoter solely under microaerobic conditions, while the K. oxytoca NifA activated it irrespective of the oxygen condition. These observations suggest that oxygen sensitivity is an intrinsic property of the A. lipoferum NifA.

Amino Acid Sequence↗
PubMed1997

Effect of bed rest immobilization on metabolic turnover of bone and bone mineral density.

Immobilization induces abnormal bone metabolism and severe decalcification of bone. To investigate the effect of middle-term immobilization on bone metabolism, we studied 10 young healthy males and females during bed rest for 20 days. Bone mineral density (BMD) rapidly decreased in both lumbar and metacarpal bones. No bone showed consistent BMD alterations, partial increase and partial decrease, and both lumbar and metacarpal bone showed similar rapid BMD change. Urinary excretion of pyridinoline tended to slightly increase by day 10, and to decline by day 20 (mean +/-SE: 34.2 +/-7.4, 26.3+/-4.6 nmol day-1, respectively). Neither alkaline phosphatase (isoform III) nor tartrate-resistant acid phosphatase, changed, suggesting that in the early stage of immobilization bone matrix in some part might increase or be resorbed without any activation of osteoblast or osteoclast, resulting in rapid calcification or decalcification, respectively.

Absorptiometry, Photon↗