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Biomedical subjects

K Wada

Publications and source records attributed to K Wada.

At least 127 records · Page 7Linked to original sources

Differential effects of social isolation upon body weight, food consumption, and responsiveness to novel and social environment in bombesin receptor subtype-3 (BRS-3) deficient mice.

The effects of social isolation on body weight gain, food consumption, and responsiveness to novel and social environment were assessed in an animal model for obesity, bombesin receptor subtype-3 (BRS-3) deficient mice. In Experiment 1, body weight gain and food consumption of group- and isolation-housed wild-type and BRS-3-deficient mice were compared. In wild-type mice, group-housed animals showed greater mean body weight gain and food consumption than did the isolation-housed cohort in the early stage of the experiment, whereas in BRS-3-deficient mice, the isolation-housed mice showed greater body weight gain and food consumption than the group-housed cohort by prolonged isolation housing. In Experiment 2, isolation-housed wild-type mice exhibited increased stereotypic and vertical movements relative to group-housed subjects in a novel environment, but this effect was not observed in BRS-3-deficient mice. In Experiment 3, when social response was assessed in animals housed in isolation, BRS-3-deficient mice exhibited lower social responses than did wild-type mice. We conclude that BRS-3-deficient mice and wild-type mice are differentially affected by social isolation. These results suggest that BRS-3 expression in the CNS may affect the neural mechanisms that regulate isolation effects in wild-type animals.

Aging↗

Evaluation of posterior cerebral artery flow velocity by transcranial color-coded real-time sonography.

Using transcranial color-coded real-time sonography (TCCS), we measured peak-systolic flow velocities (PSVs) in segment P2 of 102 posterior cerebral arteries (PCAs) in 61 patients, with angiography. We divided 102 PCAs into four groups: control group (n = 70) with no significant stenotic lesions; PCS group (n = 7) with stenosis >/= 50% of P2 segment; Col (+) group (n = 13) and Col (-) group (n = 12) had occlusive lesions in the carotid system with or without collateral flow from PCA to the middle cerebral artery through the leptomeningeal anastomosis. In the PCS group, PSV (255.7 +/- 67.2 cm/s) was higher than in the other three groups (p < 0.0001). PSV was higher in the Col (+) group (127.6 +/- 31.2) than in the Col (-) (86.6 +/- 20.1) and control (83.8 +/- 24.8) groups (p < 0.001). The measurement of PSV in the P2 segment of PCA using TCCS may help to identify a significant stenosis in PCA.

Blood Flow Velocity↗

Glutamate transporters in neonatal cerebellar subarachnoid hemorrhage.

We have previously described the immunoreactivities of glutamate transporters, EAAT4 and GLAST, in the developing human cerebellum. In the present report, we demonstrate the different expression of EAAT4 and GLAST in the pathologic condition, neonatal subarachnoid hemorrhage. EAAT4 and GLAST were characteristically disturbed in the cerebellar cortices beneath the subarachnoid hemorrhage. In preterm infants with subarachnoid hemorrhage the decrease in EAAT4 immunoreactivity was more prominent than in term infants, and GLAST immunoreactivity in the inner granular cell layer decreased and reappeared later than in term infants with subarachnoid hemorrhage. Although Bergmann's glia removes glutamate from the extracellular space surrounding Purkinje cells in the early stage of hypoxic-ischemic brain damage, the reaction of EAAT4 and GLAST in the cerebellar cortex under the subarachnoid hemorrhage was decreased, and immature glia had a delayed reaction. These characteristics of glutamate transporters in immature cells may lead to cell death and olivocerebellar degeneration.

ATP-Binding Cassette Transporters↗

Modification of glial-neuronal cell interactions prevents photoreceptor apoptosis during light-induced retinal degeneration.

Prolonged or high-intensity exposure to visible light leads to photoreceptor cell death. In this study, we demonstrate a novel pathway of light-induced photoreceptor apoptosis involving the low-affinity neurotrophin receptor p75 (p75NTR). Retinal degeneration upregulated both p75NTR and the high-affinity neurotrophin receptor TrkC in different parts of Müller glial cells. Exogenous neurotrophin-3 (NT-3) increased, but nerve growth factor (NGF) decreased basic fibroblast growth factor (bFGF) production in Müller cells, which can directly rescue photoreceptor apoptosis. Blockade of p75NTR prevented bFGF reduction and resulted in both structural and functional photoreceptor survival in vivo. Furthermore, the absence of p75NTR significantly prevented light-induced photoreceptor apoptosis. These observations implicate glial cells in the determination of neural cell survival, and suggest functional glial-neuronal cell interactions as new therapeutic targets for neurodegeneration.

Animals↗

Effects of carbamazepine on acetylcholine release and metabolism.

To clarify the mechanisms of action of carbamazepine (CBZ), we investigated the effects of CBZ on acetylcholine (ACh) release and metabolism in rat striatum and hippocampus. Acute administration of effective dose of CBZ (25 mg/kg) increased both striatal and hippocampal extracellular levels of ACh, whereas a supraeffective dose of CBZ (50 mg/kg) did not affect the levels and a toxic dose of CBZ (100 mg/kg) decreased the extracellular ACh levels in both brain regions. Both acute and chronic administrations of CBZ (25 and 50 mg/kg, mg/kg per day) increased intracellular ACh levels in striatum and hippocampus. The striatal intracellular ACh levels were decreased by both acute and chronic administrations of CBZ (100 mg/kg, mg/kg per day), whereas the hippocampal intracellular ACh levels were not affected. The effective CBZ concentration did not affect cholinesterase activity, whereas supraeffective CBZ concentration reduced it weakly. Effective dose of CBZ enhanced ACh release and synthesis; however, supraeffective doses of CBZ reduced ACh release and synthesis without enhancement of ACh degradation, indicating that CBZ has biphasic effects on ACh release and synthesis. Thus, the present findings, the slight stimulation of ACh function by effective dose of CBZ, are involved, at least partially, in the antiepileptic and mood stabilizing mechanisms of action of CBZ.

Acetylcholine↗

SPK-1, a C. elegans SR protein kinase homologue, is essential for embryogenesis and required for germline development.

SR-protein kinases (SRPKs) and their substrates, serine/arginine-rich pre-mRNA splicing factors, are key components of splicing machinery and are well conserved across phyla. Despite extensive biochemical investigation, the physiological functions of SRPKs remain unclear. In the present study, cDNAs for SPK-1, a C. elegans SRPK homologue, and CeSF2, an SPK-1 substrate, were cloned. SPK-1 binds directly to and phosphorylates the RS domain of CeSF2 in vitro. Both spk-1 and CeSF2 are predominantly expressed in germlines. RNA interference (RNAi) experiments revealed that spk-1 and CeSF2 play an essential role at the embryonic stage of C. elegans. Furthermore, RNAi studies demonstrated that spk-1 is required for germline development in C. elegans. We provide evidence that RNAi, achieved by the soaking of L1 larvae, is beneficial in the study of gene function in post-embryonic germline development.

Amino Acid Sequence↗

Induction of cyclooxygenase-2 in mesothelial cells in peritonitis caused by perforated ulcers--an immunohistochemical study in humans.

BACKGROUND: Increasing evidence suggests that mesothelial cells contribute to the control of inflammation in the peritoneal cavity by secreting prostaglandins. A study has shown that cyclooxygenase (COX)-2 knockout mice die partly as a result of peritonitis. AIM: To investigate the expression and location of COX in peritonitis associated with peptic ulcer perforation. METHODS: Gastric and duodenal tissues were collected intraoperatively from nine and four patients, respectively, and immunohistochemical staining for COX-1 and COX-2 was performed. RESULTS: Histologically, all patients had severe peritonitis around the perforation sites, into which many inflammatory cells and fibroblasts had infiltrated, and reactive mesothelial cells exhibited hyperplastic change. The COX-1 protein was not detected, whereas COX-2 was abundant in reactive mesothelial cells near the perforation site and disappeared away from the site. Macrophages and fibroblasts around the perforation site also revealed immunostaining for COX-2. CONCLUSIONS: Our results showed that COX-2 protein is induced in mesothelial cells, as well as in macrophages and fibroblasts, in inflamed peritoneal tissues associated with peptic ulcer perforation, suggesting involvement of COX-2 in tissue repair.

Adult↗

The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage.

The antigen specificity of a rat monoclonal antibody TER-119 was investigated. In adult mice, TER-119 reacted with mature erythrocytes, 20-25% of bone marrow cells and 2-3% of spleen cells but not with thymocytes nor lymph node cells. In fetal haematopoietic tissues, 30-40% of d 10 yolk sac cells, 80-90% of d 14 fetal liver cells and 40-50% of newborn liver cells were reactive with TER-119. TER-119+ cells in adult bone marrow expressed significant levels of CD45 but not myeloid (Mac-1, Gr-1) or B-cell (B220) markers. Morphological examination and haematopoietic colony-forming assays for isolated TER-119+ cells revealed that TER-119 reacts with erythroid cells at differentiation stages from early proerythroblast to mature erythrocyte, but not with cells showing typical erythroid blast-forming unit (BFU-E) and erythroid colony-forming unit (CFU-E) activities. Erythroleukaemia cell lines do not express the TER-119 antigen even after stimulation with dimethylsulphoxide. TER-119 immunoprecipitated protein bands with molecular masses of 110 kDa, 60 kDa, 52 kDa and 32 kDa from erythrocyte membrane, whereas only a 52-kDa band was detected by TER-119 in Western blot analysis. Further molecular and cellular analyses indicated that the TER-119 antigen is a molecule associated with cell-surface glycophorin A but not with glycophorin A itself.

Animals↗

Successful removal of a giant skull base metastasis from hepatocellular carcinoma after direct ethanol injection: case report.

Skull metastases from hepatocellular carcinoma (HCC) are extremely rich in vascularity, which sometimes makes surgery dangerous. For minimally invasive surgery, it is very important to diminish the intratumoral vascular flow preoperatively. We report the case of a 69-year-old man with a giant skull base metastasis from HCC that was successfully removed after two sessions of direct ethanol injection into the tumor as a preoperative treatment to diminish the intratumoral vascular flow. Direct ethanol injection is a modification of percutaneous ethanol injection therapy, which is widely used in the treatment of primary HCC. In this article, we describe in detail the practical procedures and the usefulness of this treatment for a giant skull base metastasis from HCC.

Case Reports↗

Mutation in bombesin receptor subtype-3 gene is not a major cause of obesity in the Japanese.

Bombesin receptor subtype-3 (BRS-3) is one of the candidate genes of obesity. The mice lacking BRS-3 have been shown to develop mild obesity. These mice also showed hypertension and impaired glucose metabolism, supporting these mice as a good model for human obesity. We screened 104 Japanese obese men (BMI > 26.4, 26.5-44.1) to investigate whether there is any genetic defect in BRS-3 gene. The DNA fragments containing each exon of BRS-3 gene were amplified by polymerase chain reaction (PCR) and were directly sequenced. No mutation, nor polymorphism was found in the coding region of BRS-3, suggesting that mutation of this gene is not a major cause of obesity in humans.

DNA Primers↗

Impairment of mycophenolate mofetil absorption by iron ion.

OBJECTIVE: We sought to evaluate the effect of iron ion on the absorption of mycophenolate mofetil, which is an immunosuppressive agent. The pharmacokinetics of mycophenolic acid were studied. METHODS: A randomized crossover design with two phases was used. A 7-day washout period separated the two treatment conditions. In the first phase, the volunteers received 1.0 g of mycophenolate mofetil alone (study 1); in the second phase, the volunteers received 1.0 g of mycophenolate mofetil and 2 tablets of iron ion preparations concomitantly (study 2). The serum concentration of mycophenolic acid, which is a pharmacologically active metabolite, was measured by reverse-phase HPLC. RESULTS: The area under the plasma concentration-time curve from 0 to 12 hours and the maximum concentration of mycophenolic acid in study 2 were significantly less than in study 1 (area under the curve, 32.9 +/- 14.7 versus 2.92 +/- 0.883 microg x h/mL, P < .001, maximum concentration, 20.1 +/- 9.21 versus 1.30 +/- 0.367 microg x h/mL, P < .001). CONCLUSIONS: This finding shows that when mycophenolate mofetil and iron ion preparations were administered concomitantly, a remarkable decrease of mycophenolate mofetil absorption was observed. Therefore it seems to be clear that we must avoid the concomitant administration of mycophenolate mofetil and iron ion preparations.

Adult↗

Concept for organ engineering: a reconstruction method of rat liver for in vitro culture.

In the past decade, there have been remarkable advances in tissue engineering technology toward the goal of creating organoids in vitro from cells and cellular scaffolding. Indeed, tissue-engineered organoids such as skin and cartilage, each with comparatively simple architectures, are presently at the clinical stage. However, conventional tissue engineering techniques have not allowed for the reconstruction of an organoid that mimics an organ of complex architecture of abundant vascular networks. We established a method for organ engineering that can remodel a rat liver into a reconstructed organoid without separating the majority of liver cells by a continuous three-step perfusion. The liver was perfused through its vascular system with a buffered balanced salt solution to cleanse blood from the organ, with a collagenase/dispase medium to deconstruct cellular scaffolds, and with a culture medium containing collagen type I to reorganize the multicellular architecture. The reconstructed organoid was then prepared by excising the perfused liver from the rat and culturing it at 37 degrees C for 2 h. Histologically healthy parenchymal hepatocytes expressing albumin were observed in the excised organoid even after culture for 3 weeks. Furthermore, a fibroblast-implanted organoid was prepared by using a culture medium containing suspended fibroblasts in the third step of the perfusion procedure, demonstrating the efficacy of heterogeneous cells for the reconstruction of an organoid. This method may be applicable to the formation of organoids from other organs, such as kidney and spleen, each of which have abundant capillaries, and therefore the method provides a novel concept for the development of lab-grown organs, i. e., organ engineering.

Animals↗

Real-time monitoring of glutamate following fluid percussion brain injury with hypoxia in the rat.

In the present study, extracellular glutamate (Glu) was monitored in real time using an enzyme electrode biosensor following traumatic brain injury (TBI) either with or without inducing hypoxia in the rat. We also measured the cortical contusion volume at 3 days after insult by staining with 2,3,5-triphenyltetrazolium chloride (TTC). Male Sprague-Dawley rats (300-400 g) were anesthetized and then subjected to lateral fluid percussion (FP) brain injury of moderate severity (3.5-4.0 atm), using the Dragonfly device model (no. HPD-1700). The experimental animals were divided into four groups. Group 1 (n = 10) was subjected to TBI only, group 2 (n = 10) to TBI followed by 20 min of moderate hypoxia (FiO2: 10%), group 3 (n = 4) to 20 min of moderate hypoxia without TBI, and group 4 (n = 4) to sham. Seventy-two hours after the insults, the animals were sacrificed, their brains were stained with TTC, and the lesion volumes were calculated. A surge in the extracellular Glu concentration occurred immediately after TBI in groups 1 and 2. There was no significant difference between the two groups. Group 2 showed a prolonged efflux of Glu during hypoxia ( < 0.05). In group 3, Glu continued to show a mild increase. The cortical contusion volume in group 2 was significantly larger than that in group 1. To evaluate the possible involvement of apoptosis in groups 1 and 2, separate rats were sacrificed under the same procedures after 1, 6, 24, and 72 h after insult (n = 2/group). Immunohistochemical analysis demonstrated an increased number of both the cysteine protease caspase-3-positive cells at 24 h and TUNEL-positive cells at 72 h in group 2. These results suggest that TBI with moderate hypoxia induced the prolonged efflux of Glu, which thus resulted in more cortical damage due to necrosis and apoptosis.

Animals↗

Oscillation and regulation of proline content by P5CS and ProDH gene expressions in the light/dark cycles in Arabidopsis thaliana L.

The fluctuation of proline content, and protein and mRNA levels of delta1-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH), both of which are involved in proline biosynthesis and degradation, in the shoots of Arabidopsis grown in light/dark cycles were demonstrated under salt-stressed and unstressed conditions. Proline content, as well as proteins and mRNAs of these enzymes, clearly oscillated in the light/dark cycles under the stressed and unstressed conditions. A reciprocal relationship between P5CS and ProDH was observed. Protein levels of P5CS and ProDH were well synchronized with their mRNA levels, although the fluctuation of protein levels was not as significant as that of their mRNA levels. Both mRNA and protein levels of the two enzymes as well as the proline content did not oscillate under the continuous light or the dark conditions. Thus, P5CS and ProDH gene expressions seemed to be involved in light irradiation. Moreover, relative water content (RWC) in the plants oscillated in the light/dark cycles. The fluctuations of proline content in shoot reversely responded to that of RWC. It is suggested that the expression of two genes responds sensitively to a subtle change of cellular water status, and accumulated proline keeps the osmotic balance between cells and the outer environment.

Arabidopsis↗

Influence of corn oil and diet on reproduction and the kidney in female Sprague-Dawley rats.

The purpose of this study was to investigate the influence of corn oil administration on gestation, parturition, and lactation in rats, in conjunction with diets differing in composition of nutrients. Rats were divided into two groups, each fed different commercial pellets for rodents, CA-1 or CE-2, different from each other mainly in the source of protein. Female Sprague-Dawley rats in both diet groups were administered 0 (untreated control), 2, or 10 ml corn oil/kg body weight by gavage during the premating period (2 weeks), the mating period, the gestation period, and the lactation period (until day 3 of lactation). Food consumption of both the 10 ml/kg corn oil groups was significantly reduced throughout the study. Body weight gain in the 10 ml/kg corn oil group fed the CA-1 diet was significantly reduced on days 0 through 4 of lactation. Neither mating nor fertility indices were affected, and no clinical signs were observed during the gestation period in any groups. Several dams in the 10 ml/kg corn oil group fed the CA-1 diet, however, showed abnormal conditions after parturition, and three dams became moribund. Pup viability was also reduced in this group. Histopathologic examination of the kidneys of dams in the 10 ml/kg corn oil group fed the CA-1 diet revealed severe lesions in the proximal tubular epithelium, i.e., necrosis and fatty degeneration. Females in any group fed the CE-2 diet showed neither abnormal condition after parturition nor any severe lesions in the kidney. These data show that the combination of corn oil and diet with a particular constitution may cause adverse effects on the renal tubules in pregnant and/or lactating rats, suggesting that corn oil gavage as a vehicle can be a confounding factor in the reproductive toxicity studies, depending on the diet.

Animals↗

Crystallization and preliminary X-ray analysis of endopolygalacturonase SE1 from Trichosporon penicillatum.

Endopolygalacturonase SE1 from Trichosporon penicillatum has been crystallized by the hanging-drop method of vapour diffusion using ammonium sulfate as a precipitant. The crystals belong to the hexagonal space group P6(1) or P6(5), with unit-cell parameters a = b = 135.0, c = 70.7 A, gamma = 120 degrees. The calculated V(M) based on one molecule per asymmetric unit is 3.09 A(3) Da(-1). A native data set from a crystal has been collected to 2.0 A resolution on a Cu Kalpha rotating-anode X-ray source.

Crystallography, X-Ray↗

Localisation of cyclooxygenase 1 and cyclooxygenase 2 in Helicobacter pylori related gastritis and gastric ulcer tissues in humans.

BACKGROUND: Prostaglandin endoperoxide synthase/cyclooxygenase (COX) is the key enzyme in gastric mucosal protection and repair but its cellular localisation in the human stomach is still unclear. AIMS: To investigate immunohistochemically the cellular distribution of COX-1 and COX-2 proteins in the human stomach with or without gastritis or ulceration. PATIENTS AND METHODS: Tissues were obtained by surgical resection of gastric ulcers associated with perforation (n = 9) or by biopsy from Helicobacter pylori positive patients with gastric ulcers (n = 45) and H pylori negative healthy subjects (n = 15). COX expression was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), western blotting, and light and electron microscopic immunohistochemistry. RESULTS: COX-2 mRNA and protein were detected in gastric ulcer tissues but not in intact gastric mucosa. COX-1 mRNA and protein were detected in the intact mucosa. COX-2 immunostaining was exclusively localised in macrophages and fibroblasts between necrotic and granulation tissues of the ulcer bed. The percentage of COX-2 expressing cells was significantly higher in open than in closed ulcers, and in gastritis than in gastric mucosa without H pylori infection. COX-1 immunoreactivity localised in lamina propria mesenchymal cells was similar in various stages of ulcer disease and in intact gastric mucosa. Electron microscopic immunohistochemistry revealed both COX-1 and COX-2 on the luminal surfaces of the endoplasmic reticulum and nuclear envelope of macrophages and fibroblasts. CONCLUSIONS: Our results showed that COX-2 protein was induced in macrophages and fibroblasts in gastric ulcers and H pylori related gastritis, suggesting its involvement in the tissue repair process.

Adult↗

Reduced GLP-1 and insulin responses and glucose intolerance after gastric glucose in GRP receptor-deleted mice.

By applying a newly developed ELISA technique for determining biologically active intact glucagon-like peptide [GLP-1, GLP-1-(7-36)amide] in mouse, plasma baseline GLP-1 in normal NMRI mice was found to be normally distributed (4.5 +/- 0.3 pmol/l; n = 72). In anesthetized mice, gastric glucose (50 or 150 mg) increased plasma GLP-1 levels two- to threefold (P < 0.01). The simultaneous increase in plasma insulin correlated to the 10-min GLP-1 levels (r = 0.36, P < 0.001; n = 12). C57BL/6J mice deleted of the gastrin-releasing peptide (GRP) receptor by genetic targeting had impaired glucose tolerance (P = 0.030) and reduced early (10 min) insulin response (P = 0.044) to gastric glucose compared with wild-type controls. Also, the GLP-1 response to gastric glucose was significantly lower in the GRP receptor-deleted mice than in the controls (P = 0.045). In conclusion, this study has shown that 1) plasma levels of intact GLP-1 increase dose dependently on gastric glucose challenge in correlation with increased insulin levels in mice, and 2) intact GRP receptors are required for normal GLP-1 and insulin responses and glucose tolerance after gastric glucose in mice.

Animals↗