Biomedical subjects

M Kotani

Publications and source records attributed to M Kotani.

At least 37 records · Page 2Linked to original sources

PubMed2000

Expression of prostaglandin E receptor EP4 subtype in rat adrenal zona glomerulosa: involvement in aldosterone release.

We examined the role of prostaglandin (PG) E receptors in the secretion of aldosterone. PGE2 is known to exert its various biological functions by binding to PGE receptors. There are four subtypes of PGE receptors, EP1, EP2, EP3, and EP4. Among the PGE receptors EP2 and EP4 subtypes are coupled to Gs protein and stimulate adenylyl cyclase. In this study, PGE2 caused a dose-dependent increase in aldosterone production from the rat adrenal zona glomerulosa cells in vitro accompanied with an increase in intracellular cAMP concentration. A specific agonist for EP2, butaprost, did not increase the cAMP production or the aldosterone release, suggesting the possibility that EP4 mediates the secretion of aldosterone by PGE2. Northern blot hybridization analysis disclosed that EP4 gene was expressed in the rat adrenal gland but that EP2 gene was not. In situ hybridization revealed that EP4 mRNA is present abundantly in the zona glomerulosa of rat adrenal gland. These findings suggest that the PGE2-EP4 system is involved in the regulation of aldosterone secretion from the rat adrenal gland.

Aldosterone↗

PubMed2000

Magnetometric evaluation for the effects of silicon carbide whiskers on alveolar macrophages.

Alveolar macrophages are thought to play an important role in fibrogenesis in the lungs caused by various types of exposure to dust. In this experiment, we evaluated the effect of silicon carbide whiskers (SiC) on alveolar macrophages mainly by unique magnetometry and also by established methods such as lactate dehydrogenase (LDH) activity, apoptosis measurement and morphological observations. Alveolar macrophages obtained from Syrian golden hamsters by bronchoalveolar lavages were exposed in vitro to Fe3O4 for 18 hours as an indicator for magnetometry and SiC for experiments. A rapid decrease of the remanent magnetic field, so called "relaxation", was observed after cessation of an external magnetic field in macrophages phagocytizing Fe3O4 alone, while relaxation was delayed in those concurrently exposed to SiC. Release of LDH from SiC-exposed macrophages into the medium was not significantly higher than the controls, but it increased dose-dependently. Apoptosis was recognized in macrophages exposed to 60 micrograms/ml of SiC by the DNA ladder detection method and morphological observations. Electron microscopic examination revealed irregular forms of nuclei and organellas in macrophages exposed to SiC. Magnetometry, LDH release and electron microscopic observation indicated mild cytotoxicity of SiC to alveolar macrophages.

Animals↗

PubMed1999

Possible involvement of matrix metalloproteinase-9 in Langerhans cell migration and maturation.

Epidermal Langerhans cells (LC) are potent dendritic cells in the induction of primary T cell-mediated immune responses in the skin. They capture foreign Ags and migrate to regional lymph nodes to carry and present these Ags to naive T cells. We investigated the role of matrix metalloproteinase-9 (MMP-9) in LC migration using an anti-MMP-9 mAb. Intradermal injection of anti-MMP-9 mAb before rhodamine B or oxazolone painting markedly inhibited these hapten-induced decreases in LC number in the epidermis and the accumulation of dendritic cells in the regional lymph nodes, indicating that MMP-9 plays some important roles in LC migration in the induction phase of contact sensitization. Treatment with anti-MMP-9 mAb also blocked the increase in cell size, dendrite development, and the enhanced expression of MHC class II Ags in LC induced by hapten painting. In addition, intradermal injection of purified MMP-9 induced marked increases in cell size, dendrite extension, and enhanced expression of MHC class II Ags in LC. These results strongly suggested that MMP-9 is involved not only in LC migration, but also in their morphological and phenotypic maturation in the skin.

Animals↗

PubMed1999

Bone marrow-derived cells are responsible for the development of autoimmune arthritis in human T cell leukemia virus type I-transgenic mice and those of normal mice can suppress the disease.

Previously, we reported that human T cell leukemia virus type I env-pX region-introduced transgenic (pX-Tg) mice developed an inflammatory polyarthropathy associated with a development of autoimmunity. To elucidate roles of autoimmunity in the development of arthritis, the immune cells were reciprocally replaced between pX-Tg mice and non-transgenic (Tg) mice. When bone marrow (BM) cells and spleen cells from pX-Tg mice were transferred into irradiated non-Tg mice, arthritis developed in these mice. In contrast, arthritis in pX-Tg mice was completely suppressed by non-Tg BM and spleen cells. Similar results were obtained with BM cells only. After the transplantation, T cells, B cells, and macrophages were replaced completely, whereas cells in the joints were replaced partially. In those mice, serum Ig and rheumatoid factor levels correlated with the disease development, and inflammatory cytokine expression was elevated in the arthritic joints. Furthermore, involvement of T cells in the joint lesion was suggested, because the incidence was greatly reduced in athymic nu/nu mice although small proportion of the mice still developed arthritis. These observations suggest that BM stem cells are abnormal, causing autoimmunity in pX-Tg mice, and this autoimmunity plays an important, but not absolute, role in the development of arthritis in this Tg mouse.

Adoptive Transfer↗

PubMed1999

Analyses of oviductal pars recta-induced fertilizability of coelomic eggs in Xenopus laevis.

The acquisition of fertilizability in coelomic eggs of Xenopus laevis has been shown to be correlated with the physical, biochemical, and ultrastructural alterations of the egg envelope [coelomic envelope (CE)] induced during the passage of eggs through the pars recta portion of the oviduct. However, no direct evidence that the pars recta renders eggs fertilizable has yet been presented. In this study, we show that coelomic eggs are highly fertilizable when they are incubated with continuous shaking for 4 h at 15 degrees C in pars recta extract (PRE) derived from females prestimulated by pregnant mare serum gonadotropin. The PRE from pituitary-stimulated Bufo japonicus was as potent as homologous PRE in rendering Xenopus eggs fertilizable. Incubation of coelomic eggs in PRE for 30 min induced a dramatic increase in the rates of sperm binding to the envelope to a level equivalent to that exhibited by the envelope from uterine eggs (VEs). The CE-to-VE ultrastructural conversion and a 43k-to-41k hydrolysis of the envelope glycoprotein component started 5 min after, and were completed by 15 min after, the start of incubation in PRE and were accompanied by an exposure of a new N-terminal sequence typical to gp41. Thus, the biochemical and ultrastructural conversions and the sperm-binding activity of the envelope induced by PREs, although being prerequisite, were not sufficient to render coelomic eggs fully accessible to fertilizing sperm.

Animals↗

PubMed1999

Molecular basis for oviductin-mediated processing from gp43 to gp41, the predominant glycoproteins of Xenopus egg envelopes.

Acquisition of fertilizability in Xenopus coelomic eggs is correlated with the conversion from coelomic to vitelline envelope during passage of the eggs through the pars recta portion of oviduct. The conversion includes processing of a major envelope constituent gp43 of coelomic envelopes to gp41 of vitelline envelopes by a trypsin-type protease, oviductin, which is secreted from the pars recta. Our recent sequencing analyses [Kubo et al., (1997): Dev Growth Diff 39:405-411] strongly suggested that the N-terminal portion of gp41 is exposed as a result of oviductin digestion. In this study, a monoclonal antibody specific to the predicted N-terminus of gp41 was raised by immunizing mice with a synthetic N-terminal hexapeptide (QLPVSP) coupled to keyhole limpet hemocyanin. The antibody specifically reacted to gp41, but not to gp43, indicating that Gln62 is exposed as the N-terminal amino acid of gp41 by oviductin-mediated cleavage of gp43 at Arg61 in GSR61. The C-terminal sequencing of gp43 and gp41 indicated that Arg373 in GSR373 as the C-terminus of gp41 is generated by cleavage of three amino acid (WNQ) residues from the C-terminus of gp43. The resulting polypeptide moiety of gp41 has a molecular mass of 33900 Da with 312 amino acid residues. We propose that oviductin possessing the substrate specificity of GSR simultaneously digests gp43 at Arg residues in GSR61 and GSR373 to generate the N- and C-terminus of gp41, respectively.

Amino Acid Sequence↗

PubMed1999

Involvement of autoimmunity against type II collagen in the development of arthritis in mice transgenic for the human T cell leukemia virus type I tax gene.

We previously reported that transgenic mice carrying the human T cell leukemia virus type I (HTLV-I) env-pX region (pX-transgenic mice) develop rheumatoid-like inflammatory arthropathy, and suggested involvement of autoimmunity in the pathogenicity. In this report, to elucidate pathogenesis of the arthritis, we investigated arthritogenic antigens in the joints. The TCR beta-chain variable region (Vbeta) repertoires in the lymphatic organs were normal in transgenic mice, however, specific Vbeta-positive T cells were expanded oligoclonally in the affected joints, suggesting that specific antigens, but not superantigens, were involved in the expansion of these T cells. These expanded T cells had the same TCR as those of lymph node T cells reactive to type II collagen (IIC). Moreover, these mice were susceptible to IIC-induced arthritis and oligoclonal T cells of the same Vbeta specificity as that found in spontaneously developed arthritic joint accumulated in the arthritic joints after immunization with IIC. These observations show that endogenous IIC is one of the arthritogenic antigens in the joint, suggesting tolerance break to this antigen in pX-transgenic mice.

Amino Acid Sequence↗

PubMed1999

Urinary excretion of the vitronectin receptor (integrin alpha V beta 3) in patients with Fabry disease.

A renal disorder is one of the important manifestations of Fabry disease, but the details of the pathogenesis have not been clarified yet. We examined the possibility that the vitronectin receptor (VNR, integrin alpha V beta 3), one of the integrins, is involved in the progression of the renal injury in Fabry disease. We measured the urinary excretion of beta 3 originating from VNR in Fabry patients by immunoblotting analysis and enzyme-linked immunosorbent assay (ELISA). Immunofluorescent microscopic analyses for VNR and globotriaosylceramide were performed on urinary sediments from Fabry patients. Furthermore, beta 3 and vitronectin in kidney tissues were analyzed immunohistochemically. Immunoblotting analysis and ELISA showed that the urinary excretion of beta 3 originating from VNR was significantly increased in the Fabry group compared with both the pathological and healthy control groups. Immunofluorescent microscopy revealed the expression of VNR and accumulation of globotriaosylceramide in urinary sediments from the Fabry patients. Increased expression of beta 3 was observed in glomerular epithelial cells, and in Bowman's capsular epithelial layer and tubular cells, and the amount of vitronectin was moderately increased in the kidney tissues from the Fabry patients. The urinary excretion of VNR was increased, and the expression of VNR was observed in Fabry kidney tissues. The expression of VNR may be involved in the progression of the renal injury in this disease.

Adolescent↗

PubMed1999

Identification of a novel R642C mutation in Na/Cl cotransporter with Gitelman's syndrome.

Gitelman's syndrome, a variant of Bartter's syndrome, is an inherited disorder characterized by hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria, and these abnormalities have recently been linked to the thiazide-sensitive Na/Cl cotransporter (TSC) gene. We evaluated three unrelated patients affected with this syndrome whose diagnosis was made based on clinical and biochemical features. The data of clearance studies in these patients were compatible with Gitelman's syndrome. We then investigated possible mutations of the TSC gene. In one patient whose parents are consanguineous, we identified a novel missense mutation in the TSC gene, which causes alteration of arginine to cysteine at codon 642 (R642C mutation) located in the cytoplasmic tail of the product. This mutation results in the loss of an MspI site in exon 15 of the TSC gene. MspI digestion analysis of genomic DNA fragments from the family was consistent with the autosomal recessive inheritance of the disorder, and presence of this mutation correlated with the clinical manifestations. Such mutation was not detected in 47 normal healthy subjects. In the second patient, we found another missense mutation in one allele of the TSC gene, which results in alteration of arginine to glutamine at codon 955. In the third patient, no mutation causing amino acid substitution was found in the TSC gene. These results indicate that the R642C mutation in TSC is critically important for impairment of this cotransporter function and also suggest the necessity of further investigations in the genetic background of Gitelman's syndrome.

Adult↗

PubMed1999

Roles of prostaglandin E receptors in mesangial cells under high-glucose conditions.

BACKGROUND: High glucose reportedly stimulates prostaglandin (PG) E2 production and DNA synthesis in mesangial cells (MCs). However, the pathophysiological significance of PGE2 in MCs has remained unclear. METHODS: The effects of prostanoids on [3H]-thymidine uptake and cAMP production in rat MCs cultured with 5.6 mM glucose, 25 mM glucose, or 5.6 mM glucose supplemented with 19.4 mM mannitol were examined. The gene expression of PGE2 receptor (EP) subtypes in MCs was analyzed with Northern blotting techniques. RESULTS: Northern blotting indicated EP1 and EP4 gene expression in MCs. EP1 agonists and PGE2 stimulated [3H]-thymidine uptake in MCs. EP1 antagonists dose dependently attenuated high-glucose-induced [3H]-thymidine uptake, which suggests EP1 involvement, by an increase in intracellular Ca2+, in DNA synthesis of MCs. On the other hand, forskolin, db-cAMP, and 11-deoxy-PGE1, an EP4/EP3/EP2 agonist, significantly decreased DNA synthesis in MCs. These inhibitory effects are thought to be mediated via EP4 as a result of an increase in cAMP synthesis. The effects via EP4 seem to be particularly important because PGE2-induced cAMP synthesis was significantly attenuated in the high-glucose group compared with the mannitol group, in which [3H]-thymidine uptake did not increase in spite of augmented PGE2 production. CONCLUSION: The increase in DNA synthesis in MCs under high-glucose conditions can be explained, at least in part, by the high-glucose-induced inhibition of cAMP production via EP4, which augments EP1 function in conjunction with the overproduction of PGE2.

1-Methyl-3-isobutylxanthine↗

PubMed1999

In vitro toxicity of gallium arsenide in alveolar macrophages evaluated by magnetometry, cytochemistry and morphology.

Gallium arsenide (GaAs), a chemical compound of gallium and arsenic, causes various toxic effects including pulmonary diseases in animals. Since the toxicity is not completely investigated, GaAs has been used in workplaces as the material of various semiconductor products. The present study was conducted to clarify the toxicity of GaAs particles in the alveolar macrophages of hamsters using magnetometry, enzyme release assays and morphological examinations. Alveolar macrophages obtained from hamsters by tracheobronchial lavage and adhered to the disks in the bottom of wells were exposed to ferrosoferric oxide and GaAs particles. Ferrosoferric oxide particles were magnetized externally and the remanent magnetic field was measured. Relaxation, a fast decline of the remanent magnetic fields radiated from the alveolar macrophages, was delayed and decay constants were decreased dose-dependently due to exposure to GaAs. Because the relaxation is thought to be associated with cytoskeleton, the exposure of GaAs may have impaired the motor function of them. Enzyme release assay and morphological findings indicated the damage to the macrophages. Thus the cytotoxicity causes cytostructural changes and cell death. According to DNA electrophoresis and the TUNEL method, necrotic changes occur more frequently than apoptotic changes.

Animals↗

PubMed1998

Immunohistochemical characterization of transgenic mice highly expressing human lysosomal alpha-galactosidase.

Human lysosomal alpha-galactosidase predominantly hydrolyzes ceramide trihexoside. A transgenic mouse line, C57BL/6CrSIc-TgN(GLA) 1951 Rin, highly expressing human alpha-galactosidase, has been established and investigated biochemically and immunohistochemically in order to clarify the distribution of the expressed enzyme proteins and to evaluate it as a donor model of organ transplantation therapy for Fabry disease caused by a genetic defect of alpha-galactosidase. In these transgenic mice, about five copies of the transgene were integrated, and alpha-galactosidase activity was expressed in liver, kidney, heart, spleen, small intestine, submaxillary gland, skeletal muscle, cerebrum, cerebellum, bone marrow cells and serum. The enzyme activity was about 22 to 11,080-fold higher than that in non-transgenic mice. In liver, heart and kidney tissues, which are important organs for transplantation studies, sufficient amounts of alpha-galactosidase mRNAs were transcribed, and the expressed enzymes, with molecular weights of 54-60 kDa, are abundant in the liver (enzyme activity: 53,965 nmol h-1 mg-1 protein) and heart (39,906 nmol h-1 mg-1 protein), followed by in the kidney tissue (9177 nmol h-1 mg-1 protein), respectively. An immunohistochemical microscopic study clearly demonstrated the distribution of the expressed enzyme proteins in kidney and liver tissues. Highly expressed alpha-galactosidase was detected in glomerular cells, tubular cells and hepatocytes. These transgenic mice will be useful as a donor model for experimental organ transplantation, and also it will enable recurrent biopsies and long-term observation. The organ transplantation data on mice will provide us with important information.

Animals↗

PubMed1998

Change in obstructive pulmonary function as a result of cumulative exposure to welding fumes as determined by magnetopneumography in Japanese arc welders.

OBJECTIVES: To examine the effect of chronic exposure to welding fumes on pulmonary function by a direct estimate of fumes accumulated in the lungs. METHODS: The strength of the residual magnetic field of externally magnetised lungs (LMF) was used as an estimate of fumes accumulated in the lungs. The results of forced spirometry manoeuvres obtained in 143 of 153 male welders in the original sample were cross sectionally evaluated according to LMF. Seven conventional forced spirogram indices and two time domain spirogram indices were used as pulmonary function indices, and height squared proportional correction was performed when necessary. RESULTS: The distribution of LMF values was considerably skewed towards positive. There was a weak but significant positive relation between age and log transformed LMF. Obstructive pulmonary function indices correlated well with LMF. After adjustment for age and smoking, however, a significant association with LMF was only found with percentage rate of forced expiratory volume in one second (FEV1%) divided by forced vital capacity (FVC) and mu, and average component of assumed time constant distribution of lung peripheral units. Neither FVC nor vital capacity (VC), as indices of restrictive disorders, showed a significant association with LMF. Based on the results of multiple regression analyses, a 0.6% decrease in FEV1% and 0.039 unit increase in mu were expected for each doubling of LMF. CONCLUSION: Obstructive changes in pulmonary function were found to be related to level of cumulative exposure to welding fume in male Japanese arc welders after controlling for age and smoking, assuming that LMF adequately reflects accumulation of welding fumes in the lungs.

Adult↗

PubMed1998

Magnetometric evaluation for the effect of chrysotile on alveolar macrophages.

Alveolar macrophages are thought to play an important role in fibrogenesis due to asbestos exposure. In this experiment, we evaluated the effect mainly by unique magnetometry and also by conventional methods such as lactate dehydrogenase (LDH) activity measurement and morphological observations. Alveolar macrophages obtained from Syrian golden hamsters by bronchoalveolar lavages were exposed 18 hours in vitro to Fe3O4 as an indicator for magnetometry and chrysotile for experiments. A rapid decrease of the remanent magnetic field, so called "relaxation", was observed after the cessation of an external magnetic field in macrophages phagocytizing Fe3O4 alone, while relaxation was delayed in those concurrently exposed to chrysotile. Since relaxation is thought due to the cytoskeleton-driven random rotation of phagosomes containing iron oxide particles, chrysotile is considered to interfere with the cytoskeletal function of macrophages. Release of LDH from chrysotile-exposed macrophages into the medium was recognized, but it was not significantly higher than the controls. Apoptosis was negligible in macrophages exposed to chrysotile by the DNA ladder detection, the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling method and morphological observations. Electron microscopical examinations revealed early necrotic changes in macrophages exposed to chrysotile. These findings indicate that cell magnetometry detects impaired cytoskeletal function due to in vitro exposure to chrysotile.

Animals↗

PubMed1998

Inhibition of mitosis in the guinea pig thymus after ligation of draining efferent lymphatics.

Ligature of the cervical lymph duct in guinea pigs resulted in marked dilatation of the lymphatics draining the thymus and marked reduction of mitosis in the thymus. A decrease of about 50% in the number of mitoses was found in the cortex within one day after ligation and in the medulla within five days after ligation. The thickness of the cortex also decreased rapidly and markedly in direct proportion to the decrease in cortical mitosis. Mitosis in the thymus recovered within 10 days after ligation, presumably after regeneration of lymphatics or collateralization. Thickening of the walls and valves of the efferent lymphatics of the thymus after lymph congestion was also demonstrated.

Animals↗

PubMed1997

The prostaglandin E2 and F2 alpha receptor genes are expressed in human myometrium and are down-regulated during pregnancy.

Prostaglandin (PG) E2 and PGF2 alpha are believed to play important roles in the myometrial contraction and the initiation of labor. Myometrial contraction by these prostanoids is mediated mainly through EP3 and FP, which are specific receptors to PGE2 and PGF2 alpha, respectively. During normal pregnancy, uterine myometrium are relaxed until term. To explore the involvement of EP3 and FP in the myometrial relaxation during pregnancy, we examined the EP3 and FP gene expressions in nonpregnant and pregnant myometrium obtained by hysterectomy for gynecological diseases. In all samples examined, expressions of EP3 and FP genes were detected. During pregnancy, the expression of EP3 gene in human myometrium was significantly reduced, to 60% of that in nonpregnant myometrium. The expression of FP gene in human myometrium also decreased during pregnancy to 55% of that in nonpregnant myometrium. In the myometrium from the nonpregnant women taking combined oral contraceptives, the gene expressions of EP3 and FP were not significantly changed as compared to those in nonpregnant controls. The down-regulation of EP3 and FP during pregnancy may play a role in the relaxation of myometrium and thus in the maintenance of normal pregnancy in humans.

Adult↗

PubMed1997

Structural organization of the human prostaglandin EP3 receptor subtype gene (PTGER3).

Prostaglandin EP3 receptor subtype is a seven-membrane-spanning protein with multiple C-terminal tails generated by alternative mRNA splicing. We report here the structural organization of the human EP3 gene (PTGER3). The human EP3 gene spanned more than 80 kb and was composed of 10 exons separated by nine introns. Exon 1 and the 5' 180-bp portion of exon 2 (exon 2a) encoded the seven transmembrane domains and 10 amino acid residues of the cytoplasmic tail, which are common to all EP3 isoforms. The 3' 3461-bp portion of exon 2 (exon 2b) or combinations of exons 3-10 encoded the EP3 isoform-specific C termini and formed their 3'-untranslated regions by multiple fashions of alternative mRNA splicing. Exons 2b, 4, 6, and 10 contained polyadenylation sites. The EP3 gene formed nine distinct mRNAs encoding eight EP3 isoforms, two of which were novel ones tentatively designated EP3-V and EP3-VI. The transcription initiation sites of the human EP3 gene were mapped 227 to approximately 231 bp upstream of the ATG start codon. The 360-bp 5'-flanking region contained a TATA box-like sequence, a GC box, and several cis-acting regulatory elements. The present study provides insight into the molecular mechanisms underlying the prostanoid receptor family.

Alternative Splicing↗

PubMed1997

The silent period between sounds has a stronger effect than the interstimulus interval on auditory evoked magnetic fields.

Auditory evoked cortical responses, electric. N1 and magnetic N1m, increase when the interstimulus interval (ISI) increases. We assumed that the response to a tone is mainly affected by the immediately preceding ISI, by the immediately preceding pause between stimuli (PBS) and by the previous stimulus duration (PSD). These 3 values are connected by the following expression: ISI = PBS + PSD. We examined the dependence of the auditory evoked brain magnetic responses on the ISI with the constant PSD (conventional paradigm), on the PBS with the constant ISI, and on the ISI with the constant PBS. Peak latencies and peak amplitudes of the 3 components, P1m, N1m and P2m, are recorded in one block using all possible combinations of 5 PSDs (0.05, 0.5, 1.0, 1.5 and 2.0 s) and 5 ISIs (0.5, 1.0, 1.5, 2.0 and 2.5 s). Peak latencies of these 3 components do not show any significant dependence either on the PBS or on the ISI. Neither the PBS nor the ISI brings a statistically significant effect on the P1m peak amplitude. On the other hand, the N1m peak amplitude increases as either the PBS (constant ISI) or the ISI (constant PSD) increases. The regression coefficient to the PBS is more than a double of that to the ISI. Moreover, the ISI does not show any significant effect on the N1m peak amplitude when the PBS is constant. This stronger PBS effect means that the N1m peak amplitude dependence on the ISI, which has been reported in several papers using the constant PSDs, includes more dependence on the PBS. The P2m peak amplitude shows the same tendency as the N1m because of the strong correlation in peak amplitude between them.

Adult↗