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Biomedical subjects

A Kuwahara

Publications and source records attributed to A Kuwahara.

At least 37 records · Page 2Linked to original sources
PubMed2000

Stimulation of Cl(-) secretion by L-alanine oligopeptides in the mammalian large intestine.

A variety of oligopeptides are probably released within the intestinal tissue under inflammatory conditions or during peptide absorption. To examine whether some of these peptides can affect intestinal transport functions, we determined the effects of L-alanine oligopeptide on short-circuit current (I(sc)) and transmucosal conductance (G(t)) in submucosa-mucosa preparations from the mouse cecum and guinea pig distal colon in vitro in Ussing chambers. L-Alanyl-L-alanine (Ala-Ala, 10 mM) added to the serosal side increased I(sc) and G(t), giving a peak followed by a sustained phase (the peak increase in I(sc) was 45 +/-6 microA/cm(2) and the increase in G(t) was 0.55+/-0.11 mS/cm(2)). The tripeptide, L-alanyl-L-alanyl-L-alanine (Ala-Ala-Ala, 10 mM), added to the serosal side also induced increases in I(sc) and G(t) by a similar degree. On the other hand, luminal Ala-Ala, and serosal L-alanine and L-alanine (10 mM) caused significantly smaller increases in I(sc) and G(t) ( approximately 15 microA/cm(2) and approximately 0. 15 mS/cm(2), respectively). The Ala-Ala induced increase in I(sc) was partially inhibited by serosal bumetanide (0.1 mM) and mucosal 5-nitro-2-(3-phenylpropylamino)benzoic acid (0.1 mM), and largely suppressed by removing Cl(-) from the bathing solution. The increase in I(sc) was largely suppressed by serosal low Ca(2+) and tetrodotoxin, but was not affected by indomethacin. In the guinea pig distal colon, serosal Ala-Ala (10 mM) evoked a transient increase in I(sc) by 23+/-7 microA/cm(2) and an increase in G(t) by 1.2+/-0.3 mS/cm(2). These results suggest that Ala-Ala, and probably also Ala-Ala-Ala, added to the serosal side stimulated electrogenic Cl(-) secretion mainly through the activation of submucosal secretomotor neurons in the mammalian large intestine.

Alanine↗
PubMed1999

Isolation and analysis of cDNA within a 300 kb Arabidopsis thaliana genomic region located around the 100 map unit of chromosome 1.

In order to analyze the organization of genes located at the 100 map unit of chromosome 1, we screened cDNAs hybridized with approximately 300kb contiguous DNA using four P1 clones and one YAC clone. A total of 40 kinds of cDNA were isolated, and their entire sequences were determined. A comparison with the GenBank/EMBL database indicated that three of the cDNAs have been found in Arabidopsis, and that similar sequences to 18 of the cDNAs had been detected in Arabidopsis or other organisms. cDNAs were aligned on a physical map of the contiguous DNA, and the transcriptional direction of each cDNA was determined. This contiguous DNA contains a large direct repeat, which contains five genes. In addition, identical or very similar sequences to two cDNAs are located in a narrow region. Thus, a total of 50 genes were identified, and the gene density was revealed to be approximately one gene every 6kb. In addition, cDNA sequencing revealed the existence of unusual transcripts. A sequence of seven cDNAs seemed to have no significant open reading frames. Furthermore, the existence of antisense RNA and the possibility of alternative splicing were also revealed.

Arabidopsis↗
PubMed1999

NA22598, a novel antitumor compound, reduces cyclin D1 levels, arrests cell cycle at G1 phase, and inhibits anchorage-independent growth of human tumor cells.

NA22598, a novel antitumor compound isolated from a microbial cultured broth, inhibited the growth of human colon cancer DLD-1 cells in suspension cultures (anchorage-independent growth) severalfold more strongly than in substratum-attached monolayer cultures. It arrested the cell cycle progression at early G1 phase under both these culture conditions. Rb phosphorylation, cyclin D1 expression, and cdk2 activation in G1 progression were all inhibited by NA22598, but the amounts of cdk2 and p27 were not affected. Among these effects the inhibition of cyclin D1 expression was most prominent, and NA22598 was found to inhibit the synthesis of cyclin D1 without affecting mRNA expression or protein degradation. p27 binding to cdk2 was more markedly increased in suspension cultures than in attached cultures by NA22598, but the compound had no effect on total p27. Apparently, the decrease of cyclin D1 induced redistribution of p27 from the cyclin D1/cdk4 to the cyclin E/cdk2 complexes during G1 phase in the suspension cultures. Because p27 is upregulated during suspension culture, a greater amount of it was associated with cyclin E/cdk2, thus producing greater growth inhibition. An agent, like NA22598, which induces the downregulation of cyclin D1 might offer a new anticancer strategy.

Antineoplastic Agents↗
PubMed1999

Morphological relationship between serotonergic neurons and nitrergic neurons for electrolytes secretion in the submucous plexus of the guinea pig distal colon.

In the submucous plexus, double immunocytochemistry revealed that nitric oxide synthase (NOS)-immunoreactivity was found in both numerous nerve fibers and some nerve cell bodies, while 5-hydroxytryptamine (5-HT)-immunoreactivity was limited to many nerve fibers, but not any nerve cell bodies. About 30% of the total NOS positive neurons (978) had close or some contact with 5-HT positive nerve fiber, suggesting that NO may participate in the 5-HT-evoked chloride secretion.

Animals↗
PubMed1999

New therapeutic strategy for gastric carcinoma: a two-step evaluation of malignant potential from its molecular biologic and pathologic characteristics.

BACKGROUND AND OBJECTIVES: A previous study of ours indicated that platelet-derived growth factor-A (PDGF-A) mRNA expression in biopsy specimens can identify a subgroup of high-risk gastric carcinoma patients, while clinicopathologic studies have shown that lymph node involvement is an important risk factor for predicting overall survival. To identify gastric carcinoma patients at high risk for recurrence, we assessed a two-step evaluation consisting of mRNA expression of tumor growth-related factors and the histopathologic findings. METHODS: The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assay the gene expression of PDGF-A and transforming growth factor-beta1 (TGF-beta1) in 69 gastric carcinoma endoscopic biopsy specimens (prospective cohort). The corresponding gastric carcinoma surgical specimens were classified histologically. Finally, the patients' survival curves were calculated. The relationships among the mRNA expression, histopathologic findings, and survival period were analyzed statistically. RESULTS: Nodal involvement correlated with PDGF-A and TGF-beta1 mRNA expression in early and advanced carcinomas, respectively. Both PDGF-A mRNA and TGF-beta1 mRNA expression were independent preoperative prognostic indicators in advanced cases. The ratio of involved nodes (n1) to total perigastric lymph nodes dissected (percentage of involved nodes) was the most independent postoperative prognostic indicator in advanced cases. Early carcinomas were divided preoperatively into two types. Advanced carcinomas were divided preoperatively into three. These were divided again postoperatively according to the percentage of involved nodes into high- and low-malignacy groups. CONCLUSIONS: A two-step evaluation of the malignant potential of gastric carcinoma by a combination of preoperative evaluation for PDGF-A and TGF-beta1 expression and postoperative pathologic examination would yield a more accurate prognosis for patients with gastric carcinoma.

Humans↗
PubMed1999

Comparison of the right and left ventricle as a systemic ventricle during exercise in patients with congenital heart disease.

BACKGROUND: Few studies have investigated the clinical advantages of surgical correction with the morphologic left ventricle (MLV) instead of the morphologic right ventricle as a systemic ventricle (SV) in patients with congenital heart disease. METHODS: Twenty-four healthy control subjects (group A1), 6 patients with isolated congenitally corrected transposition of the great arteries (TGA) (group A2), 16 patients with TGA who had undergone an arterial switch operation (group B1), 18 patients with TGA who had undergone a venous switch operation (group B2), 9 patients with atrioventricular and ventriculoarterial discordance who had undergone a double switch operation (group C1), and 6 patients with atrioventricular and ventriculoarterial discordance who had undergone a conventional external conduit operation from the MLV to the pulmonary artery (group C2), performed treadmill exercise testing. Their heart rate (HR), oxygen uptake (VO2), and oxygen pulse (O2 pulse), which reflects individual stroke volume, were measured, and contractile function was assessed by echocardiography. RESULTS: The peak HR for the patients after a definitive operation were significantly lower than that in group A1 and was correlated with peak VO2 (r =.67, P <.0001). The peak VO2 and peak O2 pulse for the groups A2 and B2 were significantly lower than those for the groups A1 and B1, respectively. The peak O2 pulse data were strongly correlated with those of peak VO 2 (r = 0.91, P <.0001). The left ventricular ejection fraction was significantly lower in groups B1 and C1 than in group A1 and was correlated with peak VO 2 (r =.50, P <.01). No significant differences in VO2, HR, and O 2 pulse at peak exercise were observed between groups C1 and C2. CONCLUSIONS: Chronotropic incompetence and an impaired response of the stroke volume of the MRV during exercise are partly responsible for the reduced exercise capacity in groups A2 and B2 compared with groups with the MLV as an SV, and the SV function at rest is also related to exercise capacity. Superiority of the double-switch operation compared with the conventional conduit operation was not observed. A longer-term follow-up is necessary before the advantages of these 2 operations can be compared.

Adolescent↗
PubMed1999

1-Naphthol beta-D-glucuronide formed intraluminally in rat small intestine mucosa and absorbed into the colon.

UDP-glucuronosyltransferase expressed in the rat intestinal epithelial cells is important as the first barrier against chemicals. The distribution of 1-naphthol and its glucuronide formed in rat intestine was estimated by using everted intestine. Roughly 60% of the 1-naphthol added to the mucosal fluid was absorbed into the mucosa of the small intestine and colon within 30 min. Approximately 66% of the 1-naphthol absorbed in the proximal intestine was secreted intraluminally as a glucuronide, and a minimal 9% was transported into the serosal fluid as a glucuronide. In the distal intestine, approximately 34% was secreted intraluminally and 30% was transported into the serosal fluid as a glucuronide. The greatest amount of the glucuronide (37% of the absorbed 1-naphthol) was transported into the serosal fluid, whereas a minimal 7% was secreted intraluminally in the colon. In marked contrast, the colon was found to transport 1-naphthol-glucuronide from the mucosal fluid into the serosal fluid at an approximately 8-fold higher rate than that of the small intestine. These results suggest that, in the small intestine, phenolic xenobiotics are mostly glucuronidated and secreted intraluminally and that the resulting glucuronide is absorbed and transported into the serosal side of the colon.

Animals↗
PubMed1999

Motor activity of vascularly perfused rat duodenum. 1. Characteristics of spontaneous movement.

We developed an ex vivo model of arterially perfused rat duodenum to examine the motor activity of intestine. In this preparation, spontaneously occurring pressure waves with regular rhythm were observed. The oxygen consumption and motor activity of the intestine were compared at different arterial perfusion rates to determine the degree of oxygenation required to elicit spontaneous motility. Pressure waves with regular rhythm occurred at a frequency of 1 min-1 when the arterial perfusion was 3-5 mL min-1, and stopped when the perfusion rate fell below 2 mL min-1. Atropine and hexamethonium reduced the percentage motor index/10 min of pressure waves in a dose-dependent manner, and tetrodotoxin completely blocked motor activity. Acetylcholine stimulated motor activity, and this effect was not antagonized by TTX. These findings suggest that spontaneous contraction in the ex vivo perfused rat duodenum might be mediated by a cholinergic mechanism via muscarinic receptors on smooth muscle, but that noncholinergic mechanisms may also participate in this response.

Animals↗
PubMed1999

Motor activity of vascularly perfused rat duodenum. 2. Effects of VIP, PACAP27 and PACAP38.

We examined the mechanisms of effects of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase activating polypeptide (PACAP) 27 and PACAP38 on spontaneously occurring pressure waves in ex vivo perfused rat duodenum. VIP and PACAPs dose-dependently reduced the percentage motor index of pressure waves; this reduction was not prevented by atropine, hexamethonium or tetrodotoxin (TTX). VIP and PACAPs abolished acetylcholine-induced stimulation of pressure waves, even in the presence of TTX. These findings suggest that VIP and PACAPs may exert direct inhibitory effects via VIP/PACAP receptors located on smooth muscle rather than via cholinergic receptors. The inhibitory effects of VIP and PACAPs were partially antagonized by the VIP receptor antagonists VIP(10-28), suggesting that VIP and PACAPs share common receptor sites on intestinal smooth muscle. The effects of VIP and PACAPs were completely antagonized by nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine (L-NA), suggesting that NO mediates the inhibitory effects of VIP and PACAPs on duodenal motility. Furthermore, single injection of L-NA stimulated spontaneously occurring pressure waves, while VIP(10-28) did not affect them. These findings suggest that VIP/PACAPs and NO strongly interact as an inhibitory mediator on duodenal motility, but that their modes of action in doing so may differ.

Animals↗
PubMed1999

Involvement of 5-HT3 and 5-HT4 receptors in the motor activity of isolated vascularly perfused rat duodenum.

The involvement of serotonin (5-HT) receptor subtypes in motor activity of the ex vivo vascularly perfused rat duodenum was investigated. Clusters of phasic contractions (CPCs), migrating in an oral to anal direction, were obtained without any stimulation. Drug effects were evaluated by changes in different components of the pressure waves, such as motor index (MI), frequency, amplitude and duration of the CPC. The effect of 5-HT depletion on motor activity was examined in animals treated with p-chlorophenylalanine (PCPA). The MI, frequency and duration of CPC were decreased by PCPA, but the amplitude was not affected, suggesting that endogenous 5-HT may play an important role in regulation of the motor activity of the rat intestine. The importance of the 5-HT receptor subtypes in the regulation of motor activity was examined. Neither the nonselective 5-HT1 and 5-HT2 receptor antagonist, methysergide, nor the 5-HT2 receptor antagonist, ketanserin, affected motor activity. However, the 5-HT3 receptor antagonists, granisetron and azasetron, decreased percentage MI, frequency, percentage amplitude and percentage duration of CPC. The 5-HT4 receptor antagonist, SB204070, exerted both excitatory and inhibitory actions, with a higher dose (10 nM) stimulating percentage MI, frequency, percentage amplitude and percentage duration, and a lower dose (0.1 nM or 1 nM) decreasing percentage MI and percentage duration of CPC. These results suggest that endogenous 5-HT regulates the motor activity of the rat duodenum through 5-HT3 and 5-HT4 receptors, with the former mediating the stimulatory influence and the latter mediating both stimulatory and inhibitory influences.

Animals↗
PubMed1999

Medical cost for disability: a longitudinal observation of national health insurance beneficiaries in Japan.

OBJECTIVES: To measure the impact of disability on the use of medical care and its costs. DESIGN: A 1-year prospective cohort study of National Health Insurance beneficiaries in a rural Japanese community. Their physical function was examined by the Medical Outcomes Study questionnaire at the end of 1994; medical care and its costs were then monitored for 1 year. SETTING AND PARTICIPANTS: Participants were the 49,364 subjects, aged 40 to 79 years, who were beneficiaries of National Health Insurance and lived in the catchment area of Ohsaki Public Health Center, Miyagi, Japan. The subjects were mainly farmers, self-employed persons, housewives, or pensioners. MEASUREMENTS: Medical care utilization (number of outpatient visits and days of inpatient care) and the costs for each subject were obtained from National Health Insurance Claim History files. These measurements were collected from January to December 1995, and the relationship between physical functioning levels and medical costs was analyzed. RESULTS: The medical costs per capita increased with poorer physical function. Medical costs among those with limitations in performing self-care increased by 4 times in men and 3 times in women compared with those with no functional limitation. In this cohort, the 4.3% of the subjects who were dependent in self-care used 15% of the total inpatient days and 10% of the total medical costs. CONCLUSION: Treatment of patients with disability requires a huge amount of medical resources. There is an urgent need for cost-effective intervention programs for disability prevention, which could be offset against the cost for treating the disabled.

Activities of Daily Living↗
PubMed1999

Effects of activin on hormone secretion by single female rat pituitary cells: analysis by cell immunoblot assay.

We investigated the effect of activin A on secretion of LH, FSH, and prolactin (PRL) by female cultured rat pituitary cells at the single-cell level by means of the cell immunoblot assay. Anterior pituitary cells from 8-week-old female rats were preincubated with or without activin A for 24 h, after which they were monodispersed and immediately used for cell immunoblot assay. The percentages of LH-, FSH- and PRL-immunoreactive cell blots detected were 5.5, 5.3 and 43.1%, respectively, of all pituitary cells applied to the transfer membrane. The percentage of LH-secreting cells and mean LH secretion per cell did not change after treatment with activin. In contrast, activin significantly increased the percentage of FSH-secreting cells and mean FSH secretion per cell to 136.0 and 114. 5% respectively. In addition, activin significantly decreased the percentage of PRL-secreting cells and mean PRL secretion per cell to 52.2 and 72.0% respectively. These results suggest that (1) activin A has effects on female rat pituitary cells that increase not only the amount of FSH secretion per cell but also the number of FSH-secreting cells, and (2) activin A decreases both the amount of PRL secretion per cell and the number of PRL-secreting cells.

Activins↗
PubMed1998

Effect of VIP and PACAP on vascular and luminal release of serotonin from isolated perfused rat duodenum.

Effects of CCK, VIP, PACAP38, and PACAP27 on the release of 5HT into the intestinal lumen and into the portal circulation were examined in in vivo experiments of isolated rat duodenum perfused vascularly and luminally. VIP, PACAP 38 and 27 reduced the release of 5HT into the lumen but did not affect the vascular release of 5HT. These effects were not affected by the presence of atropine, hexamethonium, or TTX, suggesting that VIP, PACAP 38 and 27 exert a direct inhibitory effect on the luminal release of 5HT from the EC cells. Nitric oxide synthase inhibitor, NG-nitro-L-arginine, antagonized the inhibitory effects of VIP, PACAP 38 and 27, suggesting that nitric oxide seems to be essential to exert the inhibitory action of VIP and PACAPs on the release of 5HT into the intestinal lumen from the EC cells.

Animals↗
PubMed1998

Expression of interleukin (IL)-12 mRNA in gastric carcinoma specimens: cellular antitumor immune responses.

BACKGROUND AND OBJECTIVES: Several tumor-related antigen peptides that are recognized by autologous cytolytic T cells (CTL) have been reported. However, most human solid tumors, including gastric carcinoma, are only weakly immunogenic. In this study, we focused on interleukin (IL)-12 and interferon-gamma (IFN-gamma) as key cytokines for estimating positive cellular immune responses. METHODS: To estimate the immunogenicity of gastric carcinomas, we examined IL-12 and IFN-gamma at mRNA levels by reverse transcription-polymerase chain reaction assay (RT-PCR) in tumor specimens and adjacent nontumor specimens from 36 gastric carcinoma patients. RESULTS: IL-12 expression was detected in 12 tumor specimens and in only two adjacent nontumor specimens (P = .003). The frequency of IFN-gamma gene expression was higher in the IL-12 mRNA-positive tumor specimens than in the IL-12 mRNA-negative tumor specimens (P = .015). In the IL-12 mRNA-positive tumors, IFN-gamma expression was higher in the tumor specimens than in the adjacent nontumor specimens (P = .007). Conversely, in the IL-12 mRNA-negative tumors, IFN-gamma expression was lower in the tumor specimens than in the nontumor specimens (P = .03). Many tumor-infiltrating mononuclear cells, predominantly T cells, were found in four of the 12 IL-12-mRNA-positive tumor specimens and in none of the 24 IL-12-mRNA-negative tumor specimens (P = .008). CONCLUSIONS: These data suggest that possible immune responses against a tumor may occur at the mRNA level in approximately one-third of gastric carcinomas.

Blotting, Southern↗
PubMed1998

Light and electron microscopic studies of pituitary adenylate cyclase-activating peptide (PACAP)--immunoreactive neurons in the enteric nervous system of rat small and large intestine.

Pituitary adenylate cyclase-activating peptide (PACAP)-immunoreactive (IR) neurons in the myenteric and submucosal plexus of the rat small and large intestine were examined by immunostaining with purified polyclonal antiserum against PACAP (1-15), using both light and electron microscopy. Many PACAP-IR neuronal cell bodies and fibers were found in the myenteric and submucosal plexus. Many of the PACAP-IR fibers originated from the cell bodies of the myenteric and submucosal ganglia. The ganglia were also innervated by PACAP-IR fibers. PACAP-IR fibers penetrated both the circular and longitudinal muscle layers, confirming the previous observations indicating that PACAP neurons act as motor neurons. Ultrastructural study demonstrated that PACAP-IR nerve terminals formed synaptic contacts with PACAP-IR nerve cell bodies or dendritic processes. This observation suggests that PACAP-IR neurons innervate other PACAP-IR neurons, and that PACAP neurons work as interneurons in the enteric nervous system. PACAP-IR nerve cells received not only PACAP-positive nerve terminal input also PACAP-negative nerve terminal input. It also suggests that PACAP neurons are regulated not only by PACAP-IR enteric neurons, but also by neurons originating elsewhere. Our observations support the view that PACAP-IR neurons are involved in the control of gut motility.

Animals↗
PubMed1998

Transforming growth factor beta1 (TGF-beta1) is a preoperative prognostic indicator in advanced gastric carcinoma.

It has been generally accepted that transforming growth factor beta1 (TGF-beta1) has both negative and positive effects on tumour growth and progression. This study analysed the prognostic value of TGF-beta1 mRNA in advanced gastric carcinoma. A reverse transcriptase-polymerase chain reaction analysis (RT-PCR) was used for TGF-beta1 in endoscopic biopsy specimens from 42 advanced gastric carcinomas. Thirty specimens expressed TGF-beta1 mRNA while 12 specimens did not. The follow-up duration ranged from 4 to 37 months (mean 22.8 months). TGF-beta1-positive group demonstrated a shorter overall survival compared with the TGF-beta1 -negative group (P=0.0014). A significant correlation was also found in the 32 patients who underwent curative resection (P=0.0048). Significant correlations were found between TGF-beta1 mRNA expression and both stage (P=0.0015) and nodal involvement (P=0.0060). Multivariate analysis demonstrated that only TGF-beta1 mRNA expression (P=0.0306) was an independent prognostic factor. All of ten patients who underwent non-curative resection expressed TGF-beta1 mRNA. Expression of TGF-beta1 mRNA in gastric biopsy specimens may be an important preoperative prognostic variable for advanced gastric carcinoma.

Aged↗
PubMed1998

Effect of VIP and PACAP on basal release of serotonin from isolated vascularly and luminally perfused rat duodenum.

The effect of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating peptide-38 (PACAP-38), and PACAP-27 on the release of serotonin (5-HT) into the intestinal lumen and the portal circulation was studied by using in vivo isolated vascularly and luminally perfused rat duodenum. 5-HT levels were determined by HPLC. VIP, PACAP-38, and PACAP-27 reduced the luminal release of 5-HT but did not affect the vascular release of 5-HT. The inhibitory effect caused by VIP, PACAP-38, and PACAP-27 was not affected by either atropine, hexamethonium, TTX, or TTX plus ACh, but it was completely antagonized by the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine (L-NNA). The VIP receptor antagonist VIP-(10-28) blocked the effects of VIP, PACAP-38, and PACAP-27. These results suggest that VIP and PACAP exert a direct inhibitory effect on the luminal release of 5-HT from the enterochromaffin (EC) cells via a common receptor site on the EC cells and that this effect is mediated by NO but not by cholinergic pathways. A single injection of TTX, atropine, or hexamethonium reduced the luminal release of 5-HT, whereas a single injection of VIP-(10-28) stimulated the luminal release of 5-HT and this effect was antagonized by atropine, hexamethonium, or TTX. These results suggest that EC cells may receive the direct innervation of cholinergic neurons as well as VIP and/or PACAP neurons, with the former exerting a tonic stimulatory influence and the latter exerting a tonic inhibitory influence on 5-HT release into the intestinal lumen.

Acetylcholine↗