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Biomedical subjects

H Kawaguchi

Publications and source records attributed to H Kawaguchi.

At least 181 records · Page 10Linked to original sources

[Usefulness of continuous venous daily chemotherapy of 5-fluorouracil and low-dose cisplatin for patients undergoing noncurative surgery].

We examined the usefulness of continuous venous daily chemotherapy of 5-FU and low/I dose CDDP for patients undergoing noncurative surgery. Twelve patients with carcinoma (1 esophageal, 6 gastric, 4 colorectal carcinoma cases and 1 carcinoma case of unknown origin) underwent resection of primary lesion and postoperative adjuvant chemotherapy. The schedule for the chemotherapy was as follows: 2-7 courses of 24-hours continuous venous infusion of CDDP (5mg/day on day 1, 2, 3, 4, 5) and 5-FU (250 mg/day on day 1, 2, 3, 4, 5 or 1, 3, 5). Several patients received a bolus infusion of Epirubicin (10 mg/1 week or 30 mg/2 week). Mean total volumes of CDDP and 5-FU were 120 mg and 5.38 g, respectively. Nine cases could be evaluated. Results of the chemotherapy were 1 CR, 1 PR, 3 NC and 4 PD, and the overall response rate was 22%. Side effects were found in 6 cases, but were not so severe. One-year survival rate was 47%, and 50% survival time was 9 months. The prognosis of the patients seemed to be relatively good. These results suggest that continuous chemotherapy of 5-FU and low dose CDDP may be useful for patients undergoing noncurative surgery.

Adult↗

[A case of esophagopleural fistula successfully cured by conservative therapy after middle and lower lobectomy of right lung].

We experience a case of esophagopleural fistula successfully cured by conservative therapy after the lung cancer operation. A 46-year-old man was received middle and lower lobectomy for adenoid cystic carcinoma of the right lung. Complication of empyema associated with an esophagopleural fistula occurred on postoperative 4th day. Conservative therapy of nothing by mouth, intravenous hyperalimentation and antibiotics was started. Three thoracic drains were inserted and the thoracic irrigation of total 3,000 ml warm saline per day twice on one day was continued. The esophagopleural fistula was closed on 6th week and the patient was discharged on 11th week after the therapy start. This complication is much rare, but recent advance in the diagnostic methods seemed to increase the indication of conservative therapy in future.

Anti-Bacterial Agents↗

[The present and future of clinical physiologic tests].

The symposium on clinical physiologic tests was designed in the 44th congress of all central laboratory divisions of national university hospitals at 28, 29 May in 1997. The title of symposium was "the present and future of clinical physiologic tests" which was named by professor M. Itoh. Four symposists were nominated among the specialists on clinical physiologic tests of members of above congress. Professor M. Itoh and Professor H. Kawaguchi spoke about Erectric Cardio Graphy (ECG) and main heart function tests and Professor M. Kambe spoke about pulmonary function tests and Professor M. Okada spoke about the using methods of clinical physiologic test data in remote and distant medicine. The special feature of clinical laboratory division in Japanese large hospitals is that has clinical physiologic tests part and the clinical laboratory division of hospitals in foreign countries has no clinical physiologic tests part. So, we expect more development of clinical physiologic tests part in nearly future.

Asthma↗

[An analysis of type 2 helper T cells in patients with atopic dermatitis].

In order to know the role of type 2 helper T (Th2) cells on patients with atopic dermatitis (AD), peripheral blood mononuclear cells were taken from the patients, stimulated with immobilized anti CD3 monoclonal antibody in the presence of monensin, and stained with fluorescein isothiocyanate-labeled anti IL-2 or IL-4 monoclonal antibody, followed by phycoerythrin labeled-anti CD4 monoclonal antibody. The fluorescent intensity was detected by FACScan. The results showed that the frequency of IL-4 producing CD4+ cells from AD patients was significantly higher than that from the healthy controls, whereas that of IL-2 producing CD4+ cells from patients were the same as healthy controls. The frequency of IL-4 producing cells was well correlated to the clinical severity of the disease, but that of IL-2 producing cells did not show any significance. Neither IL-2 nor IL-4 producing CD4+ cells had any relation to the serum IgE levels and the number of eosinophils in the patients. Our results indicate dominance of IL-4 producing CD4+ cells, which means Th2 cells in the peripheral mononuclear cells in the patients with AD, especially severe cases.

Adult↗

The role of nitric oxide in the cardiovascular system.

Nitric oxide (NO) exerts various pathophysiological effects on the cardiovascular system; inhibition of platelet aggregation or leukocyte adhesion on endothelium and vasorelaxation including lethal hypotension in endotoxic shock. In spite of these significant roles of NO, its direct action on individual cardiovascular cells remains unclarified. Therefore, we have investigated the function of NO on cells which constitute vascular wall and heart, and have found this new evidence. 1) ATP increased intracellular ([Ca2+]i) in vascular endothelial cells (ECs) and decreased [Ca2+]i of adjacently cocultured vascular smooth muscle cells (VSMCs), as detected by 2-D fura-2 image analysis. 2) The [Ca2+]i reduction in cocultured VSMCs with ECs by ATP was attenuated by pretreatment of several types of NO inhibitor, whereas the NO inhibitor potentiated the [Ca2+]i elevation in ECs, suggesting that NO affects VSMCs in a paracrine manner while ECs in an autocrine fashion. 3) Physiological concentration of lysophosphatidylcholine, which is an atherogenic constituent of oxidized LDL, but not native phosphatidylcholine, acted on ECs and VSMCs like a NO inhibitor, indicating that this material attenuates NO effect and disturbs vessel relaxation in the short term. 4) Highly efficient transfection of the ecNOS gene in rat heart showed a toxic effect on individual cardiomyocytes in vivo. In conclusion, NO may exert both beneficial and harmful effects on the cardiovascular system.

Adenosine Triphosphate↗

Alteration of extracellular matrix in dilated cardiomyopathic hamster heart.

The purpose of this study was to characterize the collagen in hereditary dilated cardiomyopathic hamster hearts, and to examine the participation of the collagen in the occurrence and progression of cardiomyopathy. BIO 53.58 hamsters (5, 10, 20 weeks old) were used as the model of dilated cardiomyopathy. Flb hamsters were used as controls. The collagen content was almost constant at any age in the Flb hamsters, but increased with age in BIO 53.58 hamsters. Type III collagen increased significantly in BIO 53.58 hamsters at 10 weeks. The acetic acid solubility of collagen decreased in BIO 53.58 hamsters as the fibrosis progressed, but was unchanged in controls. Reducible crosslinks showed a tendency to decrease progressively in BIO 53.58 hamsters. There were no differences between Flb and BIO 53.58 hamsters at 5 weeks, but its expression in BIO 53.58 hamsters at 10 and 20 weeks of age increased compared to Flb controls. These findings indicate that in the early phase of cardiomyopathy the extracellular matrix of the myocardium is rich in type III collagen. In the later phase, the matrix resembles that of hard tissues, whose collagen is mainly of type I collagen and is insoluble. These data suggest that the increased collagen synthesis may impair the cardiac function in the development of cardiomyopathy.

Animals↗

Contribution of sustained Ca2+ elevation for nitric oxide production in endothelial cells and subsequent modulation of Ca2+ transient in vascular smooth muscle cells in coculture.

To elucidate the intracellular Ca2+ (Ca2+i ) transient responsible for nitric oxide (NO) production in endothelial cells (ECs) and the subsequent Ca2+i reduction in vascular smooth muscle cells (VSMCs), we administrated four agonists with different Ca2+i-mobilizing mechanisms for both cells in iso- or coculture. We monitored the Ca2+i of both cells by two-dimensional fura-2 imaging, simultaneously measuring NO production as NO2-. The order of potency of the agonists in terms of the peak Ca2+i in ECs was bradykinin (100 nM) > ATP (10 microM) > ionomycin (50 nM) > thapsigargin (1 microM). In contrast, the order in reference to both the extent of Ca2+i reduction in cocultured VSMCs and the elevation in NO production over the level of basal release in ECs completely matched and was ranked as thapsigargin > ionomycin > ATP > bradykinin. Treatment by NG-monomethyl-L-arginine monoacetate but not indomethacin or glybenclamide restored the Ca2+i response in cocultured VSMCs to the isoculture level. In ECs, when the Ca2+ influx was blocked by Ni2+ or by chelating extracellular Ca2+, all four agonists markedly decreased NO production, the half decay time of the Ca2+i degenerating phase, and the area under the Ca2+i curve. The amount of produced NO hyperbolically correlated to the half decay time and the area under the Ca2+i curve but not to the Ca2+i peak level. Thus, the sustained elevation of Ca2+i in ECs, mainly a result of Ca2+ influx, determines the active NO production and subsequent Ca2+i reduction in adjacent VSMCs. Furthermore, L-arginine but not D-arginine or L-lysine at high dose (5 mM) without agonist enhanced the NO production, weakly reduced the Ca2+i in ECs, and markedly decreased the Ca2+i in VSMCs, demonstrating the autocrine and paracrine effects of NO (Shin, W. S., Sasaki, T., Kato, M., Hara, K., Seko, A., Yang, W. D., Shimamoto, N., Sugimoto, T., and Toyo-oka, T. (1992) J. Biol. Chem. 267, 20377-20382).

Animals↗

Multichannel optical recording of neuronal network activity and synaptic potentiation in dissociated cultures from rat hippocampus.

The activity of neuronal networks formed by dissociated rat hippocampal neurons was observed with a 128-channel optical recording apparatus using an absorptive voltage-sensitive dye, RH482. Two-dimensional patterns of neural electrical events along somata and neurites in the networks were visualized as the responses to pulse stimuli applied to the somata of the presynaptic neurons by patch-clamp electrodes. Synaptic delay was analyzed from propagation delay of the responses along the neurites. Synaptic potentiation was also observed in postsynaptic responses that were amplified by a factor of 1.24 after tetanization. In contrast, presynaptic components were unaffected by the procedure. In the light of the present results, multichannel optical recording promises to promote our understanding of neuronal interactions at cellular level.

Action Potentials↗

Enhanced expression of beta-adrenergic receptor kinase 1 in the hearts of cardiomyopathic Syrian hamsters, BIO53.58.

We cloned an entire encoding sequence of beta-adrenergic receptor kinase 1 (beta ARK1) cDNA from the hearts of Syrian hamsters through reverse transcription and subsequent polymerase chain reaction. The cloned cDNA contained 2067 nucleotides coding 689 amino acids. The sequence had 95% homology to rat beta ARK1 and 90% homology to human homologue. Cardiomyopathic Syrian hamster, BIO53.58, has been used as a model animal of congestive heart failure. M-mode echocardiography confirmed that left ventricular contractility of 20-week-old BIO53.58 was markedly reduced. The expression of beta ARK1 mRNA in the hearts of BIO53.58 was significantly increased compared to control hamsters, F1b, suggesting that the enhanced beta ARK1 expression is acting as a negative feedback mechanism in order to maintain intracellular homeostasis against accelerated stimulation by catecholamines via phosphorylation of beta-adrenergic receptor.

Aging↗

Selective Isolation of DNA or RNA Using Single-Stranded DNA Affinity Latex Particles.

We developed latex particles with a diameter of about 0.22 µm on which single-stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA.

Journal Article↗

Intact insulin-like growth factor binding protein-5 (IGFBP-5) associates with bone matrix and the soluble fragments of IGFBP-5 accumulated in culture medium of neonatal mouse calvariae by parathyroid hormone and prostaglandin E2-treatment.

We examined the distribution of insulin-like growth factor binding proteins (IGFBPs) in cultured neonatal mouse calvariae. IGFBP-3 and -4 were predominantly found in the conditioned medium. IGFBP-2 was partitioned between conditioned medium and bone and extracellular matrix (BECM), while intact (31-kDa) IGFBP-5 was most abundant in BECM extracts. After treatment with parathyroid hormone (PTH, 10(-8) M) or prostaglandin E2 (PGE2, 10(-6) M), immunoreactive IGFBP-5 accumulated in the conditioned medium in a 21-kDa form which did not bind IGF-I on Western ligand blots. PTH and PGE2 did not alter the level of steady-state IGFBP-5 mRNA, nor markedly stimulate IGFBP-5 synthesis in the calvariae, and thus accumulation of 21-kDa IGFBP-5 was largely due to release from BECM. This accumulation of truncated IGFBP-5 in the conditioned medium was not dependent on osteoclastic bone resorption, since it was not blocked by calcitonin or a bisphosphonate which inhibited PTH- and PGE2-stimulated 45Ca-release. The conditioned medium from PTH- or PGE2-treated cultures degraded recombinant human IGFBP-5 into lower molecular weight fragments. Addition of IGF-1 at 10(-8) M into the culture resulted in accumulation of native 31-kDa IGFBP-5. However, even in the presence of IGF-1, the native IGFBP-5 was degraded and the 21-kDa product accumulated in the culture medium. These results suggested a possible proteolytic mechanism for 21-kDa IGFBP-5 accumulation, responsive to PTH and PGE2. Aprotinin, leupeptin, cystatin, and bestatin did not inhibit the effects of PTH and PGE2 in the cultures. The localization of IGFBP-5 in BECM and its release and proteolysis induced by PTH and PGE2 could play a role in the local regulation of bone metabolism.

Animals↗

Interleukin-4 inhibits prostaglandin G/H synthase-2 and cytosolic phospholipase A2 induction in neonatal mouse parietal bone cultures.

We have shown previously that prostaglandin (PG) production in 7-day-old neonatal mouse calvarial cultures is regulated largely by changes in prostaglandin G/H synthase-2 (PGHS-2) expression and to a lesser extent by changes in arachidonic acid (AA) release. In this study, we examined the effects of interleukin-4 (IL-4), and its interactions with other cytokines and with parathyroid hormone (PTH), on mRNA levels of PGHS-2, PGHS-1, and cytosolic phospholipase A2 (cPLA2) and on medium protaglandin E2 (PGE2) levels in calvarial cultures. IL-1 and tumor necrosis factor-alpha (TNF-alpha), both at 1-100 ng/ml, and PTH at 0.1-10 nM increased PGHS-2 and cPLA2 mRNA and medium PGE2 levels dose-dependently after 4 h of treatment. IL-6 and IL-11 at 1-100 ng/ml did not affect mRNA or PGE2 levels. IL-4 at 1-100 ng/ml decreased PGHS-2 and cPLA2 mRNA and PGE2 levels in control as well as IL-1, TNF-alpha, and PTH-stimulated cultures. The inhibition of PGHS-2 and cPLA2 mRNA expression by IL-4 (10 ng/ml) was present at 1 h, reached a maximum at 4 h, and persisted for 24 h. The effects were maintained in the presence of cycloheximide. IL-4 also decreased PGHS-2 protein levels in control and IL-1-stimulated cultures. PGHS-1 mRNA levels were not stimulated by any of the factors studied nor inhibited by IL-4. IL-4 partially inhibited control and PTH-stimulated 45Ca release from prelabeled mouse calvariae at 4 days. However, neither the inhibition of resorption by IL-4 nor the stimulation by IL-1 and PTH were altered by indomethacin (1 microM). We conclude that (1) IL-1, TNF-alpha, and PTH, but not IL-6 nor IL-11, can increase the expression of PGHS-2, cPLA2, and PGE2 production in cultured mouse calvariae; (2) IL-4 inhibits PGE2 production in both control and stimulated calvarial cultures by inhibiting PGHS-2 and cPLA2; and (3) IL-4 has an inhibitory effect on bone resorption which is independent of PG production.

Analysis of Variance↗

Regulation of NFIL-6 and IL-6 expression by basic fibroblast growth factor in osteoblasts.

We determined whether basic fibroblast growth factor (bFGF) regulated the expression of IL-6 and NFIL-6 in osteoblasts. In mouse osteoblastic MC3T3-E1 cells, bFGF (10(-8) M) increased NFIL-6 mRNA 2-fold at 30 minutes and 3-fold at 2 h. IL-6 mRNA was increased by bFGF 10(-8) M after 1 h. IL-6 protein was detectable in control cultures but was significantly increased by bFGF (10(-8) M) at 4 h. Immunofluorescence analysis of MC3T3-E1 cells showed primarily cytoplasmic and perinuclear NFIL-6 staining in control cultures while bFGF-treated cells showed increased NFIL-6 staining at 2 and 4 h. Western blotting revealed that bFGF increased NFIL-6 protein at 2 h. In calvarial mouse osteoblasts, bFGF 10(-8) M induced IL-6 mRNA as early as 1 h and significantly increased IL-6 protein levels as early as 2 h. In conclusion, bFGF stimulates IL-6 and NFIL-6 mRNA in osteoblasts. The increase in NFIL-6 mRNA was associated with increased NFIL-6 protein. The increase in IL-6 mRNA was also associated with increased IL-6 protein. We propose that activations of NFIL-6 and IL-6 may be important mediators of the effects of bFGF on bone cells.

Animals↗

Lack of evidence for an increase in interleukin-6 expression in adult murine bone, bone marrow, and marrow stromal cell cultures after ovariectomy.

Interleukin-6 (IL-6) has been implicated as a mediator of postmenopausal bone loss. In vitro studies of bone and bone marrow cells have suggested that estrogen regulates bone turnover by controlling the production of IL-6, a potent stimulator of osteoclastogenesis and bone resorption. To investigate this hypothesis in an in vivo model, we examined the effect of ovariectomy or estrogen replacement on IL-6 mRNA and protein expression in adult mouse bone and bone marrow in vivo and in marrow stromal cell cultures. Eight-week-old CD-1 mice were sham-operated (SHAM), ovariectomized (OVX), or ovariectomized and subcutaneously implanted with 21-day slow-release pellets containing 10 micrograms of 17 beta-estradiol (O + E). Placebo pellets were implanted in the SHAM and OVX mice. Uterine weights at 1, 2, or 3 weeks after surgery were significantly decreased (68-76%) in OVX animals compared with SHAM or O + E. In mice sacrificed at 1 or 3 weeks after surgery, we found by nonquantitative reverse transcribed polymerase chain reaction (RT-PCR), that SHAM, OVX, and O + E calvariae (CALV) constitutively expressed IL-6 mRNA. In contrast, IL-6 mRNA was either barely detectable or absent in the tibia (TIB) and bone marrow (BM). In the mice sacrificed 3 weeks after surgery, we determined by quantitative RT-PCR that IL-6 mRNA in the CALV from the OVX and O + E groups were decreased by 81 and 92%, respectively, compared with SHAM. IL-6 protein levels in the flushed bone marrow (BMSups) were detectable and were not significantly different among the groups. In bone marrow cells that were cultured for 1 week, basal levels of IL-6 protein were low and did not differ significantly among the SHAM, OVX, or O + E groups sacrificed 1, 2, or 3 weeks after surgery. After the addition of hrIL-1 alpha, IL-6 protein levels increased 100- to 1300-fold over control. IL-6 levels in cells from animals sacrificed 2 weeks after surgery were significantly lower in the hrIL-1 alpha-stimulated OVX and O + E groups than in hrIL-1 alpha-stimulated SHAM cell cultures. In conclusion, in this model we could find no increase in IL-6 production with in vivo estrogen withdrawal in calvaria, long bones, bone marrow, or marrow stromal cell cultures. If increases in IL-6 expression are involved in the effects of estrogen withdrawal on bone, the magnitude of these changes are relatively small and below the limits of detection of the assays that we employed.

Animals↗

Water-in-oil emulsion containing oxine for the collection of traces of copper(II) in waterPreliminary stud.

An oil type emulsion containing tiny encapsulated droplets of hydrochloric acid has been used for the concentration of traces of copper(II) ions in water. Milligram quantities of oxine and a non-ionic surfactant (Span-80) were dissolved in 5-10 ml of chloroform and mixed vigorously with 3 ml of 1 mol/l hydrochloric acid by ultrasonic irradiation. The resulting water-in-oil type emulsion was gradually added to 50-500 ml of water sample and dispersed by stirring as numerous small globules (0.1 to 0.5 mm in diameter). The copper diffused through the chloroform layer into the small droplets of hydrochloric acid, which occurred quantitatively in the sample of pH 3-10. After separating the emulsion by sedimentation, it was demulsified by heating to segregate the aqueous and organic phases. The copper in the aqueous phase was successfully determined by GFAAS. The emulsion method allows to perform both extraction and back-extraction more easily and rapidly than the conventional liquid-liquid extraction method.

Journal Article↗

A point mutation in exon 7 of the C1-inhibitor gene causing type I hereditary angioedema.

The polymerase chain reaction and nucleotide sequence analysis have been used to characterize a point mutation in the seventh exon of one allele of the C1-inhibitor gene in a family with type I hereditary angioedema. A single base change (C-->T) at nucleotide 1482 in C1-inhibitor converted the codon for Gln-339 to a premature translation termination codon, TAG. Family studies suggest that this mutation is responsible for type I hereditary angioedema in a studied pedigree.

Amino Acid Sequence↗

Band neutrophil count and the presence and severity of coronary atherosclerosis.

It has been consistently shown that the total blood leukocyte count is an independent risk factor for coronary artery disease. Few studies, however, have addressed the relation between differential leukocyte counts and coronary artery disease. We investigated the relation of total and differential leukocyte counts to angiographically determined coronary atherosclerosis. The study included 486 subjects (335 men, 151 women) who underwent coronary angiography for suspected coronary artery disease. Band neutrophil count was significantly positively related to coronary atherosclerosis (p = 0.004) after adjustments for age, sex, body mass index, cigarettes per day, serum total cholesterol, and hypertension. Although sex and age-adjusted total blood leukocyte count was significantly positively related to coronary atherosclerosis (p = 0.04), this relation did not reach significant levels (p = 0.08) after adjusting for other risk factors. The positive association with band neutrophil counts was at least as strong as that with serum total cholesterol concentrations. This study indicates that band neutrophil counts serve as an independent risk factor for coronary atherosclerosis.

Adult↗