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Biomedical subjects

S Kawashima

Publications and source records attributed to S Kawashima.

At least 235 records · Page 13Linked to original sources

Role of sex steroids on the survival, neuritic outgrowth of neurons, and dopamine neurons in cultured preoptic area and hypothalamus.

Morphological sex differences in some discrete brain regions are thought to be developed by the influence of circulating androgen during the perinatal period. In order to know whether the effect of androgen is a direct one, cells derived from neonatal rat preoptic area (POA) and/or hypothalamus were cultured in a serum-free medium, and the effects on survival, process growth of neurons, and dopaminergic function were examined. When the POA cells were exposed to testosterone (T), neuronal survival was greater than the controls, and the frequency distribution of total process length and the number of process branchings were significantly deviated from the controls. Estradiol-17 beta (E2) and 5 alpha-dihydrotestosterone had less significant effects on these parameters than T. Moreover, the addition of T increased the dopamine (DA) contents of cells and medium DA in the hypothalamic culture. E2 also greatly increased DA content of the medium. These steroids failed to alter the DA levels in the POA cell cultures. These results generally conform to the notion of previous investigators that T has direct effects on the expansion of dendritic elements of the POA and the development of sex difference in the hypothalamic DA neurons, although the effect of T after conversion to E2 cannot be excluded.

Animals↗

Effects of gonadotropin-releasing hormone (GnRH) on the cytodifferentiation of gonadotropes in rat adenohypophysial primordia in organ culture.

The effects of gonadotropin-releasing hormone (GnRH) on the development of gonadotropes were investigated by the use of organ culture and by means of immunocytochemistry and radioimmunoassay. Pituitary primordia from rat fetuses were cultured in a medium with or without 10(-9) M GnRH during the first 24 h of culture. The ratio of the number of immunoreactive LH cells to the total number of cells in the explants derived from 13.5-day fetuses was increased by the GnRH treatment after 6 or 8 days of culture, while the total number of cells was not altered. LH released into the medium and LH content of explants were not affected by the GnRH treatment. Subsequent treatment with 10(-9) M GnRH for 4 h after 7 days of culture resulted in a marked release of LH, accompanying a significant decline in LH content, in both explants exposed or unexposed to the first GnRH treatment. However, the former explants contained a lower amount of LH than the latter explants. The present results indicate that pituitary primordia at 13.5 days of gestation are capable to respond to GnRH, and that GnRH is effective in stimulating the responsiveness of gonadotropes to GnRH during early pituitary cytodifferentiation.

Aging↗

Immunohistochemical double-labeling study of gonadotropin-releasing hormone (GnRH)-immunoreactive cells and oxytocin-immunoreactive cells in the preoptic area of the dwarf gourami, Colisa lalia.

The distribution of gonadotropin-releasing hormone (GnRH)-immunoreactive (ir) cells in the preoptic area (POA) of the dwarf gourami, Colisa lalia, was immunohistochemically studied. These neurons form cell columns on both sides of the common ventricle, and their axons project to the pituitary gland by forming distinct bundles. Also examined was the distribution of isotocin (IST) cells in the POA by using an anti-oxytocin (OXT) serum which has been proven to crossreact with IST. These two kinds of immunoreactive cells were distributed quite similarly in the POA. However, by using an immunofluorescence double-labeling method on thinner sections we found that a population of small IST cells in the ventral POA were also immunoreactive to GnRH, but that large IST cells in the dorsal POA were not immunoreactive to GnRH, and small GnRH-ir cells in the most ventral POA were not immunoreactive to the OXT antiserum. In the pituitary gland, GnRH-ir fibers were found in both the neurohypophysis and proximal pars distalis, but IST fibers were found only in the neurohypophysis.

Animals↗

Subcellular distribution of calpain and calpastatin immunoreactivity and fodrin proteolysis in rabbit hippocampus after hypoxia and glucocorticoid treatment.

Rabbits were subjected to hypoxia (5% O2) for up to 90 min and allowed to recover for a maximum of 4 days. Hippocampus homogenate was assayed for fodrin breakdown product (BDP). After separation into a nuclear and mitochondrial fraction (NMF), a membrane and microsomal fraction (MMF), and a cytosolic fraction (CF), samples were assayed for mu-calpain, m-calpain, and calpastatin immunoreactivity. Calpain and calpastatin immunoreactivity decreased in the NMF and CF but increased in the MMF during hypoxia and short-term recovery. This translocation occurred in parallel with the increase in fodrin BDP. Because the increase in the MMF was not large enough to explain the decrease in the other two fractions, it was assumed that the translocation and activation was accompanied by a reduction in the total amounts of calpains and calpastatin. Glucocorticoid pretreatment (beta-methasone, 0.4 mg x kg-1 x day-1) for 7 days produced a decrease in the ratio of activated mu-calpain in all three fractions in nearly all samples before, during, and after hypoxia, compared with untreated animals. Glucocorticoid pretreatment also prevented the increase in fodrin BDP that occurred in untreated animals during hypoxia and short-term recovery, indicating impairment of calpain activation.

Animals↗

A partial connectin cDNA encoding a novel type of RSP motifs isolated from chicken embryonic skeletal muscle.

A partial cDNA encoding 811 amino acids of connectin (titin), a giant elastic protein of muscle (3,000 kDa), was cloned from a chicken embryonic skeletal muscle cDNA library using antibodies to muscle connectin. The encoded product was the C terminal segment of connectin. The predicted sequences consisted of 5 type II motifs (immunoglobulin C2 type) separated by 5 interdomain insertions. One interdomain insertion had significant homology (RSP) to KSP repeats found in human cardiac C-terminal connectin and another had a high sequence homology to porin (67.7%; 31 amino acids).

Amino Acid Sequence↗

Calpain-calpastatin interactions in epidermoid carcinoma KB cells.

We examined the activation of mu-calpain in human epidermoid carcinoma KB cells following a rise in intracellular calcium concentration using antibodies specifically recognizing different activation states of mu-calpain. KB cells possess calpastatin activity in large excess of calpain activity as analyzed by ion exchange HPLC. Stimulation of the cells with a calcium ionophore, ionomycin, caused production of the autolytic intermediate form (M(r) = 78 k) of mu-calpain derived from the preautolysis form (80 k), while the fully autolyzed postautolysis form (76 k) remained below detectable levels at all times. The appearance of the autolytic intermediate paralleled limited proteolysis of the membrane-associated calpastatin fractions (110 k and 106 k); the resulting fragments (68 k and 45 k) were released into the cytosol. Both the production of the autolytic mu-calpain intermediate and the limited proteolysis of calpastatin in cell lysates in the presence of calcium were inhibited by a synthetic calpastatin peptide, indicating that proteolysis of calpastatin was indeed catalyzed by calpain and that the autolytic intermediate may have exerted the proteolytic activity. Furthermore, mu-calpain autolysis and calpastatin degradation, upon ionomycin treatment, were both augmented by epidermal growth factor (EGF). These results suggest that calpastatin serves not only as an inhibitor but also as a substrate for calpain at cell membranes and that intracellular conditions associated with the cell cycle may affect the activation of mu-calpain.

Amino Acid Sequence↗

The toxic effect of Alzheimer amyloid protein precursor overexpressed in the neuroblastoma cell line NB-1 on neurite outgrowth.

The neuroblastoma cell line NB-1 is induced to start neurite outgrowth by dibutyryl cyclic AMP (Bt2cAMP). To study the function of Alzheimer amyloid protein precursor (APP), a stable cell line overexpressing APP was established by cDNA transfection. The APP cDNA was driven by the cytomegalovirus early gene promoter. The transformant underwent degeneration as well as neurite outgrowth in the presence of Bt2cAMP, which also increased the amount of APP mRNA and protein. The expressed APP was mainly processed in the secretory pathway and few amyloidogenic fragments were observed. Hence, not only the beta/A4 protein but also the overexpressed APP itself might be neurotoxic.

Amyloid beta-Protein Precursor↗

Transforming growth factor-beta 1 potentiated alpha 1-adrenergic and stretch-induced c-fos mRNA expression in rat myocardial cells.

Since transforming growth factor-beta 1 (TGF-beta 1) has been recently shown to be expressed in the heart by mechanical stretch and ischemic injury, we examined the modulation of c-fos mRNA expression and amino acid uptake by TGF-beta 1 in rat myocardial cells. Pretreatment with TGF-beta 1 potentiated norepinephrine (NE)-induced and stretch-induced (+10% and +20% elongation, for 30 minutes) c-fos mRNA expression by 2.2-fold, whereas TGF-beta 1 alone did not induce c-fos mRNA expression in Northern blot analysis. NE-induced [14C]phenylalanine uptake was also potentiated with TGF-beta 1 pretreatment. The effect of TGF-beta 1 on the NE action was not blocked by propranolol but by prazosin. The protein kinase C activators (12-O-tetradecanoylphorbol 13-acetate [TPA], phorbol 12,13-dibutyrate, and 1-oleyl-2-acetyl-rac-glycerol) induced c-fos mRNA expression, which was also potentiated by TGF-beta 1. Cycloheximide (protein synthesis inhibitor) could not suppress the TGF-beta 1 actions. In nonmuscle cells, TGF-beta 1 modified neither adrenergic nor TPA-induced c-fos mRNA expression. These data suggested that TGF-beta 1 potentiated the c-fos mRNA expression and amino acid incorporation by modification of the alpha 1-adrenergic and stretch-activated protein kinase C pathway. This mechanism did not require protein synthesis.

Amino Acids↗

Augmented receptor-mediated Ca2+ mobilization causes supersensitivity of contractile response to serotonin in atherosclerotic arteries.

We have previously reported that atherosclerotic arteries obtained from Watanabe heritable hyperlipidemic (WHHL) rabbits exhibit a marked increase of contractile response to serotonin (5-hydroxytryptamine [5-HT]) and ergonovine and that these augmented contractile responses to specific agonists may play an important role in the pathogenesis of vasospasm. In the present study, we investigated whether supersensitivity to 5-HT in atherlosclerotic arteries was due to an increase in 5-HT receptor-mediated Ca2+ mobilization or to an increase in Ca2+ sensitivity of the contractile elements. We measured simultaneously both isometric tension and [Ca2+]i in fura 2-loaded aortic smooth muscle strips from control and WHHL rabbits. Muscle tension in the high K+ (72.7 mmol/L)-stimulated states and [Ca2+]i in both resting and high K(+)-stimulated states did not differ between control and WHHL rabbits. In atherosclerotic aortas from WHHL rabbits, the dose-response curves of both tension and [Ca2+]i for 5-HT were shifted to the left at lower threshold concentrations and one-half maximally effective dose. The maximum response of contraction produced by 5-HT in WHHL rabbits was augmented compared with that in control rabbits (123 +/- 17% versus 33 +/- 7% of the 72.7 mmol/L K(+)-induced contraction, P < .001). The maximum response of [Ca2+]i produced by 5-HT was also augmented in WHHL rabbits compared with control rabbits (29 +/- 4% versus 10 +/- 0.9% of the 72.7 mmol/L K(+)-induced [Ca2+]i, P < .001). In contrast, the responses of contraction and [Ca2+]i to phenylephrine were similar between control and WHHL rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of intracoronary infusion of adenosine on cumulative deterioration of regional myocardial wall motion induced by repeated brief periods of coronary occlusion in anesthetized dogs.

The present study was performed to evaluate the effects of the intracoronary infusion of adenosine on regional wall motion in the canine heart subjected to repeated brief coronary occlusion and reperfusion. In anesthetized open-chest dogs, six episodes of five-minute coronary occlusion followed by a fifteen-minute reperfusion were performed. Animals received infusions of either adenosine or saline (control) into the coronary artery during the initial five minutes of each reperfusion. Changes in regional wall motion were studied by means of ultrasonic crystals. The authors prolonged the duration of hyperemic flow at each reperfusion in a third group by means of a roller pump to examine whether the increase in coronary flow during reperfusion itself might affect the process of the deterioration of regional wall motion. In the control dogs, wall thickening decreased progressively after each reperfusion. However, in the adenosine-treated animals, regional wall motion fifteen minutes after the sixth reperfusion did not differ from baseline. Prolongation of hyperemic flow did not improve the progressive decline in regional wall motion. Thus, adenosine prevented the cumulative decrease in the regional wall motion of the myocardium treated with a series of repeated transient coronary occlusions, most likely by a mechanism unrelated to its vasodilating action.

Adenosine↗

A clinical and pathologic study of a large Japanese family with Machado-Joseph disease tightly linked to the DNA markers on chromosome 14q.

The gene locus for Machado-Joseph disease (MJD) has been mapped to chromosome 14q by linkage analysis, mainly using a single large Japanese family. We studied the clinical and neuropathologic findings of this family with MJD, comparing them with those of spinocerebellar ataxia 1 (SCA1) and spinocerebellar ataxia 2 (SCA2) families. The pedigree included 30 affected persons in 125 members of five generations. Neurologic examination of 21 patients revealed that dystonia, difficulty in eyelid opening, slowness of movements, bulging eyes, and facial-lingual fasciculation-like movements or myokymia are characteristic of this MJD family, although these three autosomal dominant spinocerebellar degenerations have several neurologic signs and symptoms in common. In contrast with SCA1 and SCA2, degeneration of the subthalamopallidal system and relative sparing of the olivocerebellar system were the main neuropathologic features of MJD.

Adult↗

Impaired endothelium-dependent vasodilatory responses in hindlimb blood flow in dogs with congestive heart failure.

The vasodilatory response is known to be diminished in chronic heart failure. In the present study, we used dogs with chronic heart failure to investigate whether endothelium-dependent vasodilation in the resistance vessels in the hindlimb is decreased in chronic heart failure, and if so, to determine if endothelium-derived relaxing factor (EDRF) is involved in the mechanism. We induced heart failure in dogs by continuous rapid ventricular pacing. Under anesthesia, an internal carotid artery-femoral artery bypass system was established and perfusion pressure to the femoral artery was kept constant. Changes in hindlimb blood flow in response to various concentrations of acetylcholine (ACh), adenosine 5'-diphosphate disodium salt (ADP) and nitroglycerin (NTG) were examined in dogs with and without heart failure. The vasoconstrictive responses to L-NG-monomethyl L-arginine monoacetate (L-NMMA) were also studied. In dogs with heart failure, vasodilatory responses to both of the endothelium-dependent vasodilators, ACh and ADP, were decreased, while there were no differences in the responses to the endothelium-independent vasodilator NTG. The vasoconstrictive response to L-NMMA was diminished in heart failure. Thus, endothelium-dependent vasodilation in hindlimb resistance vessels is impaired in dogs with congestive heart failure. This impairment may be related to diminished EDRF/NO production or release in these vessels.

Acetylcholine↗

Enhanced cell proliferation by hyperprolactinemia in both exocrine and endocrine pancreas in mice.

Effects of hyperprolactinemia induced by ectopic anterior pituitary grafting on the pancreas were studied in male SHN mice. After pituitary grafting, the weight of pancreas rapidly increased. A similar increase in pancreatic weight was observed during lactation, a condition associated with elevated prolactin levels. Results of DNA and protein assays revealed that the increase in pancreatic weight in both pituitary-grafted and lactating mice was mainly due to the increase in the cell number, because the total DNA content per pancreas was greater in these mice than the controls. An increase in fluid volume or hypertrophy of cells also contributes to the weight increase; in contrast, the DNA and protein contents per unit tissue weight decrease. The rate of DNA synthesis determined by 5-bromo-2'-deoxyuridine labeling was higher both in acinar cells and islet B cells in pituitary-grafted mice than in the controls. Thus, hyperprolactinemia stimulates cell proliferation in exocrine pancreas as well as endocrine islets. The effect of prolactin seems to be indirect on acinar cells, because only B cells showed prolactin immunoreactivity in the mouse pancreas. In addition, insulin might not be a mediator of the prolactin effect on acinar cells, because the serum insulin level in pituitary-grafted mice failed to show any change.

Animals↗

The significance of right ventricular ischemia in reflex bradycardia in humans.

Heart rates (HR) were measured before (control HR) and after (ischemic HR) balloon inflation during percutaneous transluminal coronary angioplasty in 96 patients with effort angina pectoris. The delta HR, defined as (ischemic HR-control HR), in the right coronary artery occlusion was significantly smaller than that in the left anterior descending artery or left circumflex artery occlusion. Among right coronary artery occlusions the delta HR in segment 1 occlusion was significantly smaller than that in segment 2 or segment 3 occlusion. It is suggested that the ischemia of the right ventricle plays an important role in producing a reflex bradycardic response in humans.

Angina Pectoris↗

[A case of MELAS associated with prosopagnosia, topographical disorientation and PLED].

We report a case of mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) associated with prosopagnosia, topographical disorientation, and periodic lateralized epileptiform discharge (PLED) on electroencephalography (EEG) in a 23-year-old right-handed man. The first MELAS attack occurred on March 1, 1991, while the patient was drinking. Magnetic resonance imaging (MRI) revealed a lesion of abnormal intensity in the left occipital lobe. The second attack occurred on October 1, 1991. This time, the major symptoms were visual loss of acute onset, nausea, and vomiting. EEG examination showed transient PLED. MRI revealed a new area of abnormal intensity in the right occipital lobe, lingual gyrus, fusiform gyrus and the posterior part of the parahippocampal gyrus. During the clinical course of the patient, prosopagnosia and topographical disorientation appeared. There have been few reports of MELAS associated with prosopagnosia, topographical disorientation, and PLED. However, MELAS attacks tend to occur in the cortex of the occipital lobe. We therefore believe that these neuropsychological symptoms and PLED are likely to be associated with MELAS.

Adult↗

[Pharmacokinetic, bacteriological and clinical studies on cefozopran in children].

Pharmacokinetic, bacteriological and clinical studies on cefozopran (CZOP) were performed in children. The results were as followed: 1. A total of 13 patients were treated with CZOP. The tested dose was 20 mg/kg (50 mg/kg in maxillary sinusitis), and the drug administered via intravenous bolus injection or 30-minute intravenous drip infusion 3 times daily, for 3-11 days. Clinical efficacies of CZOP in 13 patients with bacterial infections (11 with pneumonia, 1 with otitis media and 1 with maxillary sinusitis) were evaluated as excellent in 13 with an efficacy rate of 100%. Any adverse reactions or abnormal laboratory test results were not observed in any patients. Fourteen causative strains were found in 10 patients. Streptococcus pneumoniae in 4 cases out of 4, Streptococcus pyogenes in 1/1, Moraxella (Branhamella) catarrhalis in 3/3, Haemophilus influenzae in 4/6 were eradicated. 2. Pharmacokinetic studies. The mean serum concentration immediately after intravenous drip infusion over 30-minute of 20 mg/kg was 39.1 micrograms/ml (range: 25.6-52.5 micrograms/ml). The mean urinary recovery rate over 8 hours after administration was 49.0% (range: 45.2-51.8%). Based on the above results and the broad spectrum and great antibacterial activity of CZOP, it is considered that CZOP is a promising antibiotic usable as a single agent for the primary therapy of acute bacterial infections ranging from mild to severe in children.

Bacteria↗

[A case of anti-phospholipid antibody syndrome associated with typical mesangiolysis].

An 18-years-old female was admitted to our hospital because of severe edema, that had developed within one week, and massive proteinuria. She has been given treatment for mild proteinuria and Raynaud's phenomenon. Before the edema developed, she complained of recurrent attack of migraine associated with scintillating scotoma. At admission, she was suffering from butterfly erythema, hydropericardium and ascites. The results of tests for antinuclear antibodies and ENA antibodies were negative, but those for BFP and IgG-anticardiolipin antibody were positive, and the platelet and complement levels were low. Renal biopsy revealed typical mesangiolysis without any sign of hyperplastic nephritis. Electron microscopy demonstrated the deposition of microfibrillar structures in the mesangial areas. She was diagnosed as having SLE-antiphospholipid antibody syndrome. After the administration of prednisolone 40mg/day, the edema, Raynaud's sign and proteinuria disappeared. Migraine was improved by the additional use of warfarin, and the platelet count was normalized. These findings suggest that in this case, acute nephrotic symptom developed due to hemangioendothelial injury caused by anti-phospholipid antibody.

Adolescent↗