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Biomedical subjects

Y Masuda

Publications and source records attributed to Y Masuda.

At least 253 records · Page 14Linked to original sources

Activation of protein kinase C prevents induction of apoptosis by geranylgeraniol in human leukemia HL60 cells.

In a previous study, we showed that geranylgeraniol (GGO) is a potent inducer of apoptosis in human leukemia cells, including HL60 promyelocytic leukemia cells. The present study describes the effects of activators of protein kinase C (PKC) on GGO-induced apoptosis in various lines of leukemia cells. Both 12-O-tetradecanoylphorbol-13-acetate (TPA) and diacylglycerol (DG) inhibited the GGO-induced morphological changes that are characteristic of apoptosis and the DNA fragmentation. Similar effects were observed with other lines of human and murine leukemia cells such as ML1, U937, M1 and P388. Flow cytometric analysis also revealed that both TPA and DG prevented GGO-induced DNA degradation in a dose-dependent manner. These inhibitory effects of TPA and DG were antagonized by inhibitors of PKC such as H-7 and staurosporin, and by amiloride, an inhibitor of Na+/H+ antiporter. In contrast to the inhibitory effects of TPA and DG on GGO-induced apoptosis, 4alpha-TPA, which is unable to activate PKC, failed to prevent GGO-induced DNA fragmentation. However, the selective activator of PKC-beta, 12-deoxyphorbol 13-phenylacetate 20-acetate, significantly inhibited GGO-induced DNA fragmentation. Our results suggest that PKC, and in particular the PKC-beta isoenzyme, might be involved in the process of GGO-induced apoptosis.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Nuclear and cytosolic calcium signaling induced by extracellular ATP in rat kidney inner medullary collecting duct cells.

Using a laser scanning confocal microscopy of fluorescent Ca2+ indicator (Fluo-3-AM) the spatiotemporal Ca2+ dynamics in cultured kidney inner medullary collecting duct cells was investigated. In response to extracellular ATP (100 microM), nuclear (Fln) and cytosolic (Flc) fluorescence intensity increased simultaneously. UTP similarly increased Fln and Flc, but ADP and AMP did not. A ratio between Fln and Flc was about 1.06 +/- 0.03 at rest and increased 1.71 +/- 0.02 at the peak of stimulation (n = 74). In Ca(2+)-free condition, ATP increased Fln and Flc with a smaller peak intensity, but the peak ratio was similar (1.52 +/- 0.03, n = 70). Faster time resolution of 100 ms in line scanning mode did not detect the delay between nuclear and cytosolic Ca2- responses. Our results indicate that nuclear Ca2+ was not diffused from the cytoplasm and that it may be directly released from the nuclear envelope, a possible Ca2+ store.

Adenosine Triphosphate↗

Antagonist activity of [Thr18,gamma-methylleucine19]endothelin-1 in human endothelin receptors.

Receptor binding and antagonist properties of endothelin-1 analogues, [Thr18,gamma-methylleucine19]endothelin-1, [Thr18,Leu19]endothelin-1 and [Thr18,cyclohexylalanine19]endothelin-1, were investigated using cloned human endothelin ET(A) and ET(B) receptors expressed in Chinese hamster ovary cells. Among them, [Thr18,gamma-methylleucine19]endothelin-1 had a high affinity for endothelin ET(A) and ET(B) receptors with respective Kd values of 300 and 110 pM and had no agonist activity on the stimulation of arachidonic acid release in endothelin ET(A) and ET(B) receptor-expressing cells. [Thr18,gamma-methylleucine19]Endothelin-1 had potent antagonist activity in endothelin-1-induced arachidonic acid release in endothelin ET(A) and ET(B) receptor-expressing cells with respective pA2 values of 8.2 and 8.5. In an inositol phosphates accumulation assay, [Thr18,gamma-methylleucine19]endothelin-1 also exhibited potent antagonist activity for endothelin ET(A) and ET(B) receptors with respective pA2 values of 8.0 and 8.4. In conclusion, [Thr18,gamma-methylleucine19]endothelin-1 acts as a potent and nonselective antagonist with no agonist activity for cloned human endothelin ET(A) and ET(B) receptors.

Animals↗

Galactose-inducible expression systems in Candida maltosa using promoters of newly-isolated GAL1 and GAL10 genes.

The GAL1 and GAL10 gene cluster encoding the enzymes of galactose utilization was isolated from an asporogenic yeast, Candida maltosa. The structure of the gene cluster in which both genes were divergently transcribed from the central promoter region resembled those of some other yeasts. The expression of both genes was strongly induced by galactose and repressed by glucose in the medium. Galactose-inducible expression vectors in C. maltosa were constructed on low- and high-copy number plasmids using the promoter regions of both genes. With these vectors and the beta-galactosidase gene from Kluyveromyces lactis as a reporter, galactose-inducible expression was confirmed. Homologous overexpression of members of the cytochrome P-450 gene family in C. maltosa was also successful by using a high-copy-number vector under the control of these promoters.

Amino Acid Sequence↗

A mixed mesodermal tumor of the cecum: report of a case.

We herein report the case of a 69-year old woman presenting with an abdominal mass, who was found to have a mixed mesodermal tumor (MMT) of the cecum. Imaging studies and endoscopic investigations were consistent with the diagnosis of a nonepithelial malignant tumor of the cecum. On laparotomy, a knuckle-sized firm mass involving the cecum was noticed. As a result, a right hemicolectomy was performed. Pathological examinations, including immunohistochemical staining, resulted in the diagnosis of mesodermal mixed tumor, homologous type. The patient was advised to undergo postoperative chemotherapy but she did not comply. She has been followed up as an outpatient and is still alive 1.5 years after the operation.

Aged↗

Evaluation of the three-dimensional clinostat as a simulator of weightlessness.

Concerns regarding the reliability of slow-and fast-rotating uni-axial clinostats in simulating weightlessness have induced the construction of devices considered to simulate weightlessness more adequately. A new three-dimensional (3-D) clinostat equipped with two rotation axes placed at right angles has been constructed. In the clinostat, the rotation achieved with two motors is computer-controlled and monitored with encoders attached to the motors. By rotating plants three-dimensionally at random rates on the clinostat, their dynamic stimulation by gravity in every direction can be eliminated. Some of the vegetative growth phases of plants dependent on the gravity vector, such as morphogenesis, are shown to be influenced by rotation on the 3-D clinostat. The validity of 3-D clinostatting has been evaluated by comparing structural parameters of cress roots and Chara rhizoids obtained under real microgravity with those obtained after 3-D clinostatting. The parameters analyzed up to now (organization of the root cap, integrity and polarity of statocytes, dislocation of statoliths, amount of starch and ER) demonstrate that the 3-D clinostat is a valuable device for simulating weightlessness.

Plant Physiological Phenomena↗

Laparoscopic-assisted colon surgery by abdominal wall lifting with newly developed lifting bars.

BACKGROUND: The aim of the study is to evaluate the efficacy of laparoscopic-assisted colon surgery by lifting the abdominal wall with newly developed lifting bars. METHODS: We have made and used two kinds of lifting bars: type I and type T. Two I-type lifting bars are used in transverse colectomy and right hemicolectomy. One I-type lifting bar and one T-type bar are used in sigmoid colectomy and low anterior resection. After the intestine is dissected and the mesenterium is treated under laparoscopy, a small laparotomy wound about 4 to 6 cm long is made, and the intestine is pulled out of the body for extracorporeal anastomosis. RESULTS: The mean operating time was 153.8 +/- 51.9 min, and no particular complications were noted. CONCLUSIONS: Since postoperative pain is mild and postoperative recovery is rapid, this method is considered to be an effective surgical procedure.

Abdominal Muscles↗

New indices of ischemic heart disease and aging: studies on the serum levels of soluble intercellular adhesion molecule-1 (ICAM-1) and soluble vascular cell adhesion molecule-1 (VCAM-1) in patients with hypercholesterolemia and ischemic heart disease.

It is known that the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on the surface of vascular endothelial cells is closely related to the formation of early atherosclerotic lesions. In this study, serum soluble ICAM-1(sICAM-1) and soluble VCAM-1(sVCAM-1) were determined by sandwich ELISA both in normal healthy individuals (n = 114) and in patients with hypercholesterolemia (HC, n = 112) or ischemic heart disease (IHD, n = 38) to clarify the significance of the soluble forms of the adhesion molecules in the development of atherosclerotic diseases. IHD patients, not HC patients, showed significant elevation of sICAM-1, but not of sVCAM-1, compared with controls in age and sex-matched subjects. In addition, multiple linear regression analysis showed that sICAM-1 was correlated only to the presence of IHD but not to age and lipids. Multiple logistic regression analysis revealed that sICAM-1 was the most powerful independent predictor of the presence of IHD. On the other hand, sVCAM-1, not sICAM-1, was positively correlated to age. Multiple linear regression analysis showed that age was the most powerful independent predictor of the level of sVCAM-1. These data suggest that sICAM-1 and sVCAM-1 are useful as indices of clinical manifestations of atherosclerosis and aging, respectively.

Adult↗

Bufalin reduces the level of topoisomerase II in human leukemia cells and affects the cytotoxicity of anticancer drugs.

When human leukemia HL-60 cells were treated with 10(-7) M bufalin, the amounts of both topoisomerase (topo) II alpha and II beta and the activity of topo II decreased markedly and were almost undetectable 18 h after the start of treatment. The level of topo II mRNA started to decrease immediately after the start of treatment with bufalin, with a subsequent decrease in the amount of topo II alpha protein. These changes preceded the fragmentation of DNA, a typical feature of apoptosis. The results suggest that bufalin caused a marked decrease in the steady-state level of topo II alpha mRNA, which led to a decrease in the amount and activity of the enzyme and to the induction of apoptosis. A reduction in the level of topo II alpha by bufalin was also observed in other lines of human leukemia cells such as ML1 and U937. The results were exploited to potentiate the effects of cisplatin and retinoic acid (RA) on HL-60 cells: pretreatment of HL-60 cells with 10(-7) M bufalin for 6 h increased the inhibitory effects of cisplatin and RA on cell growth and enhanced the induction of cell death.

Antineoplastic Agents↗

Evaluation of facial palsy by moiré topography index.

We investigated the usefulness of moiré topography for evaluating facial nerve function in 51 patients with facial palsy and 10 normal volunteers. This method visualizes the shape of objects in three dimensions. We devised three moiré indexes as a simple method of quantifying the severity of facial palsy: the nasolabial groove moiré index, the oral angle moiré index, and the inner canthus moiré index. We compared the results obtained by the moiré indexes with findings obtained using the House-Brackmann grading system and found that they were highly correlated. These three moiré indexes included all the standard factors of the House-Brackmann grading system, allowing us to develop the total moiré index.

Adolescent↗

Inhibitory effect of monatepil maleate on acyl-CoA:cholesterol acyltransferase activity in the liver of cholesterol-fed Japanese monkeys.

We have previously demonstrated that monatepil maleate, AJ-2615, a new calcium antagonist endowed with alpha1-adrenoceptor blocking property, has antiatherosclerotic and plasma lipid-lowering effects in Japanese monkeys fed on a cholesterol-rich diet. To clarify the mechanisms on plasma lipid-lowering action, we investigated the effect of monatepil maleate in these monkeys on hepatic acyl-CoA:cholesterol acyltransferase (ACAT) activity. Both ACAT activity and esterified cholesterol content in the livers of monkeys fed on a cholesterol-rich diet for 6 months significantly increased about 7- and 16-fold, respectively, as compared with those in monkeys fed on a standard diet. Monatepil maleate (30 mg/kg/day for 6 months, orally) inhibited the increases of ACAT activity and esterified cholesterol content by 51% and 71%, respectively. In in vitro experiments, monatepil maleate inhibited ACAT activity in a concentration-dependent manner, whereas it did not affect 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity. A kinetic analysis revealed that monatepil maleate was a noncompetitive type inhibitor of ACAT. Hepatic ACAT activity was significantly correlated to hepatic esterified cholesterol content (r = 0.775, P < .0001), to plasma very low density lipoprotein (VLDL) content (r = 0.765, P < .0001) and to plasma total cholesterol content (r = 0.573, P < .005) in the monkeys. These results suggest that ACAT-inhibiting effect of monatepil maleate plays an important role in the reduction of hyperlipidemia.

Animals↗

Analysis of two pharmacologically predicted endothelin B receptor subtypes by using the endothelin B receptor gene knockout mouse.

1. This study was performed to clarify whether the endothelin (ET) receptor subtypes mediating two pharmacologically heterogeneous response to ETH receptor agonists in normal mice are the product(s) of a single ETB receptor gene. 2. Vasodilator responses to sarafotoxin S6c (S6c) in the thoracic aorta and contractile responses to ET-1 and IRL1620 in the stomach were examined in tissues from normal and ETB receptor gene knockout mice, in the absence and presence of an ETA receptor antagonist, BQ-123, or an ETA/ETB receptor antagonist, PD142893. 3. In the normal mouse aorta precontracted with phenylephrine, S6c (0.1-100 nM) caused concentration-dependent relaxations (pD2 = 8.4). BQ-123 had no effect on these responses. However, PD142893 almost abolished the relaxations induced by 0.1-300 nM S6c. 4. In aortae taken from ETB receptor gene knockout mice, S6c up to 1 microM failed to cause relaxations, confirming that ETB receptors are involved in mediating this response. 5. In normal mouse gastric fundus, 0.1 nM-1 microM ET-1, S6c or IRL1620 caused dose-dependent, BQ-123-insensitive contractions, which were much more resistant to PD142893 than S6c-induced relaxations of the aorta. The pD2 values for S6c in the absence and presence of PD142893 (10 microM) were 8.12 +/- 0.11 and 7.70 +/- 0.11, respectively. 6. In the gastric fundus of the ETB receptor gene knockout mouse, S6c and IRL1620 caused no contractions. ET-1 (0.1 nM-1 microM) caused contractions sensitive to both BQ-123 and PD142893, indicating that only ETA receptors mediate ET-1-induced contractions of the knockout mouse gastric fundus. 7. Since both the PD142893-sensitive vasodilator response of the aorta and the PD142893-resistant contractile response of the gastric fundus to S6c were completely absent in the ETB receptor gene knockout mouse, we conclude that the two pharmacologically heterogeneous responses to S6c are mediated by receptors derived from the same ETB receptor gene.

Animals↗

Metabolism of 3,3',4,4'-tetrachlorobiphenyl via sulphur-containing pathway in rat: liver-specific retention of methylsulphonyl metabolite.

1. Single doses of 3,3',4,4'-tetrachlorobiphenyl (TCB) were administered intraperitoneally to the male Wistar rat for metabolism studies. 2. Seven sulphur-containing metabolites of TCB were isolated from faeces, in addition to previously reported 4-hydroxy-3,3',4',5-tetrachlorobiphenyl and 5-hydroxy-TCB. Major sulphur-containing metabolites were 5- and 6-methylthio-TCBs, and 6-methylsulphonyl-3-methylthio-3',4,4'-trichlorobiphenyl. 3. The faecal excretions of hydroxy metabolites, methylthio metabolites and unchanged TCB accounted for 7.1, 0.5 and 0.3% of the dose respectively within 4 days after administration. 4. 5-Methylsulphonyl-TCB was detected and selectively retained in liver. The concentration ratio of 5-methylsulphonyl-TCB and unchanged TCB in liver was 1:4. 5. Following administration of 5- and 6-methylsulphonyl-TCBs to rat, 5-methylsulphonyl-TCB was localized in liver, whereas 6-methylsulphonyl-TCB was rapidly biotransformed to 6-methylsulphonyl-3-methylthio-3',4,4'-trichlorobiphenyl and excreted in the faeces.

Adipose Tissue↗

Normal human fibroblasts immortalized by introduction of human papillomavirus type 16 (HPV-16) E6-E7 genes.

This report demonstrates that normal human fibroblasts can be immortalized by the introduction of HPV-16 E6-E7 genes. We designed zinc-inducible expression plasmids with HPV-16 E6, E7 or both. Each plasmid was introduced into normal human fibroblasts (TIG-3 cells) using lipofection methods. Only transfectants with the HPV-16 E6-E7 zinc-inducible expression plasmid, which were cultured in medium supplemented with 100 microM ZnSO4, overcame crisis and could be cultured over 200 population doubling levels (PDLs). These cell lines showed the reactivation of telomerase after crisis, and morphological alterations were also observed.

Cell Culture Techniques↗

Quantitative analysis of cyclin D1 messenger RNA expression in head and neck squamous cell carcinomas.

Cyclin D1 is thought to play a critical role in the G1/S phase transition of the cell cycle. Amplification of this gene has been reported in several types of human neoplasms including breast, lung, esophageal, and head and neck tumors. In this study, we have analyzed the relative level of expression of cyclin D1 messenger RNA (mRNA) in fresh specimens of head and neck squamous cell carcinoma (HNSCC), and investigated the concordance of the overexpression of cyclin D1 mRNA with gene amplification. Levels of cyclin D1 mRNA were analyzed by a modified method of competitive reverse transcription-polymerase chain reaction and levels of cyclin D1 gene amplification were evaluated by Southern blot hybridization in a series of 23 matched normal mucosas and HNSCC. Overexpression of cyclin D1 mRNA was observed in 10 of 23 cases (43.5%) of HNSCC, ranging from 2 to 50-fold higher than the normal control. Twelve of 23 cases could be evaluated by Southern blot hybridization, and gene amplification was found in only 2 of 12 cases (16.7%). These findings suggest that cyclin D1 plays an important role in tumorigenesis of HNSCC, and gene amplification is not one of the major mechanisms for overexpression of cyclin D1.

Aged↗

Protective effect of zonisamide, an antiepileptic drug, against transient focal cerebral ischemia with middle cerebral artery occlusion-reperfusion in rats.

PURPOSE: The antiepileptic effects of zonisamide (ZNS) have been well documented experimentally and clinically. The purpose of this study was to examine whether ZNS reduces cerebral damage after transient focal ischemia in rats. METHODS: Ischemia was induced by a transient occlusion of the left middle cerebral artery (MCA) with a 3-0 nylon monofilament for 90 min. Neurological evaluation was performed by measuring the event of neurological deficit of the contralateral forepaw and hindpaw at 10 min and 1 day after MCA occlusion (MCAo). Brain infarct size was determined by measuring triphenyltetrazonium chloride-negative stained area of the serial brain sections 1 day after MCAo. RESULTS: The pre- or postischemic treatment with ZNS [(10-100 mg/kg p.o.), 30 min before and 4 h after or 15 min and 4 h after the occlusion] markedly reduced cerebral damage in the ipsilateral hemisphere and the neurological deficit induced by transient ischemia. The reducing effect on the damage was observed in the cortical and subcortical regions. Preischemic treatment with carbamazepine (CBZ 60 mg/kg p.o. twice 30 min before and 4 h after MCAo) tended to reduce the cerebral damage and neurological deficit, but the lower dose (20 mg/kg p.o. twice) did not. Valproate (VPA 1,000 mg/kg p.o. twice) also had no effect. CONCLUSIONS: ZNS at the anticonvulsant dose, unlike CBZ and VPA, ameliorated the brain infarction and the event of neurological deficit after transient focal cerebral ischemia. These data suggest that ZNS has therapeutic potential in protecting against ischemic cerebral damage, such as stroke.

Animals↗

[Clinical study on penicillin insusceptible/resistant Streptococcus pneumoniae in the elderly patients].

Clinical features of respiratory infection in the elderly with penicillin insusceptible (31 cases) and resistant (7 cases) Streptococcus pneumonia (PSSP/PRSP) are compared to those with penicillin sensitive S. pneumoniae (PSSP) (29 cases). Incidence of bacteremia and pneumonia was higher in the PSSP group. PISP/PRSP tend to be isolated from patients with bronchitis underlaid with chronic pulmonary disease without statistic significance. Efficacy of the penicillins and 1st and 2nd generation cephem was satisfactory except in only one case of pneumonia with PISP which needed an alternative choice to the 3rd generation cephem. Now a day the degree of resistance is not so high and the available antibiotics are sufficient for the treatment of pneumococcal infection in the elderly patients. However, the wide use of oral cephems and certain new quinolones which do not have enough activity against Streptococcus pneumoniae may increase resistance. In which case, continuous surveillance and clinical caution against this resistant strain is necessary.

Aged↗