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Biomedical subjects

K Kishi

Publications and source records attributed to K Kishi.

At least 109 records · Page 6Linked to original sources

Age-related changes of gene expression in mouse kidney: fluorescence differential display--PCR analyses.

We used a fluorescence differential display--PCR (FDD-PCR) technique to analyze the genes expressed in mouse kidneys collected at nine different developmental stages ranging from 3 days to 15 months after birth. We found ten genes that were age-dependent and differentially-expressed in the kidneys during our experimental period. We confirmed by comparative RT-PCR that of the ten cDNAs, seven showed reproducible age-dependent expression. Four of the nucleotide sequences of these cDNA clones, had high homology with known genes (fibronectin, soluble guanylyl cyclase alpha-1 subunit, cytosolic aldehyde dehydrogenase and mitochondrial DNA), and three with expressed sequence tags of unknown genes. The FDD-PCR method was very useful for detecting new age-related genes expressed differentially in the mouse kidney.

Aging↗

Gi-mediated translocation of GLUT4 is independent of p85/p110alpha and p110gamma phosphoinositide 3-kinases but might involve the activation of Akt kinase.

Activation of phosphoinositide 3-kinase (PI-3K) is essential for insulin-stimulated translocation of GLUT4 and glucose transport in insulin target tissues. A novel p110gamma PI-3K was reported to be activated by G(i)-coupled receptors via Gbetagamma subunits. We asked whether the stimulation of G(i)-coupled receptors would trigger GLUT4 translocation and glucose uptake by the activation of Gbetagamma-dependent p110gamma PI-3K. We find that this translocation and glucose uptake can be induced by the ligand stimulation of G(i)-coupled alpha(2A) adrenergic receptor and fMet-Leu-Phe receptor in cells stably expressing these receptors. The noradrenaline ('noradrenaline')- and fMet-Leu-Phe-stimulated GLUT4 translocations were abolished by pretreatment with pertussis toxin. Pretreatment with wortmannin or genistein also inhibited the G(i)-mediated GLUT4 translocation. On ligand stimulation of these two kinds of G(i)-coupled receptor, although there was a slight increase in PtdIns(3,4,5)P(3) production, activation of either the p85/p110alpha PI-3K or Gbetagamma-dependent p110gamma PI-3K was not observed even in Chinese hamster ovary cells stably overexpressing exogenous p101/p110gamma. The G(i)-mediated GLUT4 translocation was accompanied by activation of the serine-threonine kinase Akt; the inhibitory effects of pertussis toxin, wortmannin and genistein on G(i)-mediated GLUT4 translocation paralleled their inhibitory effects on Akt activation. In contrast, the activation of some other G(i)-coupled receptors, such as prostaglandin EP3alpha receptor and platelet-activating factor receptor, did not cause either pertussis-toxin-sensitive translocation of GLUT4myc or activation of Akt kinase. These results indicate that the ligand stimulation of some G(i)-coupled receptors triggers GLUT4 translocation that occurs independently of p85/p110alpha-type and p110gamma-type PI-3Ks but might involve the activation of Akt kinase.

Adrenergic alpha-Agonists↗

Postmortem absorption of dichloromethane: a case study and animal experiments.

A case of accidental death after occupational exposure to an atmosphere containing dichloromethane (DCM) is reported. The concentrations of DCM in the blood and tissues of a 40-year-old man who died while observing an industrial washing machine filled with DCM vapour were blood 1660 mg/l, urine 247 mg/l, brain 87 mg/kg, heart muscle 199 mg/kg and lungs 103 mg/kg which are 3-7 times higher than previously reported fatal levels. The body was left undiscovered in the machine filled with DCM vapour for about 20 h. The present study was designed to determine whether all the DCM detected in the tissues and body fluids had been inhaled while alive using rats as the experimental model. The concentrations of DCM in the tissues and body fluids of a rat that died from DCM poisoning and was left for 20 h in a box containing DCM vapour were the same as those in the tissues and body fluids of a rat that had died from an injected overdose of barbiturates and had then been placed in the DCM box in a similar manner. Moreover, the concentrations of DCM in the tissues and body fluids of the carcasses that were exposed to the DCM vapour increased gradually throughout the period of exposure. These findings imply that DCM is able to penetrate the tissues and body fluids of rat carcasses through a route other than inhalation such as through the skin.

Adult↗

Geranylgeranylacetone stimulates mucin synthesis in cultured guinea pig gastric pit cells by inducing a neuronal nitric oxide synthase.

Nitric oxide (NO) has been considered to play an important role in the regulation of blood flow, mucosal integrity, and mucus production in the stomach. We investigated the stimulatory actions of epidermal growth factor (EGF) and a cytoprotective compound, geranylgeranylacetone (GGA), on mucin synthesis in guinea pig gastric pre-pit cells, maintained in a serum-free culture system. GGA increased [3H]glucosamine uptake and the accumulation of mucus granules positive for galactose oxidase-Schiff reaction in the cells. This stimulatory action of GGA was equivalent to that of EGF, but GGA did not stimulate the cell growth. Both EGF and GGA increased the release of NO degeneration products, NO2- and NO3-. The [3H]glucosamine uptake was completely inhibited by the non-selective NO synthase (NOS) inhibitors, N(G)-nitro-L-arginine and N(G)-monomethyl-L-arginine, and it was only partially inhibited by a more selective inhibitor for inducible NOS isoform (iNOS), aminoguanidine. Northern blotting with a cDNA probe for rat iNOS, and Western blotting with a polyclonal antibody against iNOS, demonstrated that GGA did not up-regulate the iNOS mRNA expression nor induce its protein. In contrast, GGA and EGF induced neuronal NOS, but not endothelial NOS, which was confirmed by immunoblot analyses with antibodies against these constitutive NOS isoforms. Thus, the present experiments suggests that GGA, as well as EGF, stimulates mucin synthesis at least in part through an NO-dependent pathway, leading to an increase in the integrity of the gastric mucosa.

Animals↗

Macrolides and clindamycin suppress the release of Shiga-like toxins from Escherichia coli O157:H7 in vitro.

We investigated the effects of antimicrobial agents fosfomycin (FOS), cefdinir (CDIN), levofloxacin (LEVX), rokitamycin (ROK), roxithromycin (ROX), and clindamycin (CLI) on the release of Shiga-like toxins (SLTs) by enterohaemorrhagic Escherichia coli (EHEC). EHEC was cultured for 14 h in the presence of ROX, ROK or CLI at sub-minimum inhibitory concentrations (subMICs) of 1.56-6.25 mg/l, followed by assay of the level of SLTs in the supernatants using cytotoxicity assay and reversed passive latex agglutination method. Exposure to ROX, ROK or CLI reduced the amount of released SLTs compared with untreated control cultures (P<0.05). These agents however, did not decrease the number of viable EHEC, indicating the importance of bactericidal agents. When the bacteria was exposed to CDIN, FOS or LEVX, the level of SLTs in the culture supernatant increased with the destruction of bacterial cells in the order of CDIN, FOS, LEVX. When 0.5 mg/l of LEVX was added to cultures with or without pretreatment using ROX, ROK, or CLI, the release of SLTs was reduced by this pretreatment (P<0.05). These results may have clinical implications for the treatment of EHEC infection.

Animals↗

Japanese population data for two STR loci, HumTPO and HumLPL.

The two short tandem repeat (STR) systems, HumTPO and HumLPL, were investigated in blood samples obtained from approximately 800 unrelated Japanese individuals living in seven geographically different areas of Japan. Neither deviation from a Hardy-Weinberg equilibrium nor significant difference between the allele distributions was found among the seven Japanese populations in the two STR systems. These findings indicate that there is a general uniformity for both the STR loci in the Japanese population.

Journal Article↗

A case of glandular odontogenic cyst associated with ameloblastoma: correlation of diagnostic imaging with histopathological features.

The glandular odontogenic cyst (GOC) is a rare odontogenic cyst. There has only been one reported case of the simultaneous presence of GOC and ameloblastoma. The radiographic features of GOC are well established but the MR findings have not been described. We report a case of GOC associated with ameloblastoma with special reference to the correlation of the diagnostic imaging with the histopathological features.

Ameloblastoma↗

Two cases of polyostotic eosinophilic granuloma.

We report two cases of polyostotic eosinophilic granuloma (EG). Both plain radiographs and CT showed diffuse osteolytic lesions which suggested malignant tumors. Although EG was polyostotic, the prognosis was relatively good. Both cases developed new lesions over a follow-up period of 4-5 years and therefore further long-term review is needed.

Adult↗

Comparative study of the temporomandibular joint space in maximum intercuspation and canine edge-to-edge positions in deep bite and non-deep bite subjects.

In order to study the relationship between morphological properties of the TMJ and the mathematically predicted force distribution in the TMJ, the geometric distribution of the joint space on top of the condylar surfaces was evaluated. Seven deep bite subjects and seven non-deep bite subjects were used, and the joint space in the maximum intercuspation and canine edge-to-edge positions was analysed in each group. Standardized images of the TMJ were taken with computed tomography (CT) using a mandible positioner. The comparative ratio of joint space (CRJS) between the maximum intercuspation and the canine edge-to-edge positions was calculated and summarized on the map of the mean morphological outline of the condyle. On the balancing-side condyle, the CRJS of small value was distributed mainly on the anterior edge of the condyle in the non-deep bite group, whereas, in the deep bite group, it was distributed on the centre of the condyle. On the working-side condyle, the small CRJS was distributed around the lateral pole of the condyle in the non-deep bite group, whereas, in the deep bite group, it was found around the posterior part of the condyle. The distribution pattern of CRJS on the upper surface of the condyle in the non-deep bite group was consistent with the results of previous studies on mathematical simulation of TMJ loading. From the results of this study, it was suggested that the distribution of mechanical loading on the TMJ in the eccentric mandibular position would be highly affected by the comparative ratio of joint space. As the comparative ratio of joint space would be highly affected by the condylar position, the analysis of the morphology of the TMJ in relation to mechanical joint loading, in a future investigation, should be carried out on specific mandibular positions in relation to the function of the TMJ to be studied.

Adult↗

Successful endoscopic resection of intramural metastasis after esophagectomy for esophageal cancer: a case report.

Carcinoma of the esophagus is often accompanied by intramural metastasis (IM) at the time of diagnosis, and the prognosis of patients with such metastasis is very poor. Here we report the case of a 60-year-old man who presented with a submucosal tumor in the wall of the cervical esophagus at 2 years after esophagectomy for thoracic esophageal cancer. The tumor was resected endoscopically and was histologically shown to be an IM of esophageal cancer. He has shown no recurrence and no metastasis during 2 years of follow-up after endoscopic resection and radiation therapy.

Carcinoma, Squamous Cell↗

A non-toxic heat shock protein 70 inducer, geranyl-geranyl-acetone, restores the heat shock response in gastric mucosa of protein-malnourished rats.

Acute gastric mucosal lesions caused by stress or noxious stimuli are important to consider in the management of critically or chronically ill patients. Protein malnutrition has been implicated as a risk factor for stress ulcer and subsequent complications in those patients. When male Wistar rats fed a 5% or 20% casein diet for 3 weeks were exposed to restraint and water-immersion stress, the low-protein diet significantly increased the ulcer index. The low-protein diet did not change the level of heat shock factor 1 (HSF1) in gastric mucosa but it did attenuate the HSF1 activation after exposure to the stress, resulting in the inhibition of HSP70 mRNA expression and HSP70 induction in gastric mucosa. HSP70 is crucial for the maintenance of cell integrity during pathophysiologic conditions; therefore the impaired HSP70 induction appeared to at least in part aggravate stress ulcer. We also tested whether a non-toxic HSP70 inducer, geranyl-geranyl-acetone (GGA), effectively improved the mucosal integrity by stimulating HSP70 induction under protein malnutrition. Intragastric administration of GGA (200 mg/kg twice a day) to the protein-malnourished rats for up to 1 week failed to stimulate the HSP70 induction. However, the administration of GGA (200 mg/kg twice a day) for 3 weeks restored HSP70 induction and induced higher resistance against stress ulcer as compared with results in vehicle-treated, normally nourished rats. Our results suggest that GGA may have a potential benefit for the prevention of stress ulcer in chronically or critically ill patients with protein malnutrition.

Animals↗

Hypothermia attenuates the vasodilatory response of pial arterioles to hemorrhagic hypotension in the cat.

UNLABELLED: We investigated the effect of hypothermia on the vasodilatory response of pial arterioles to hemorrhagic hypotension. The cranial window technique was combined with microscopic video recording in an experiment involving 20 cats anesthetized with pentobarbital. The animals were randomly assigned to either a normothermic or a hypothermic group (32 degrees C). Mean arterial pressure (MAP) was reduced in stepwise increments of 10 mm Hg (from 100 to 50 mm Hg) by blood withdrawal. The diameter of small (50-100 microm) and large (100-200 microm) pial arterioles was measured. In the normothermic group (n = 9), small and large arterioles dilated at a MAP of 60 and 50 mm Hg, and at a MAP of 70, 60, and 50 mm Hg, respectively, compared with baseline values obtained at a MAP of 100 mm Hg. In contrast, in the hypothermic group (n = 11), vasodilation of either small or large arterioles was absent. The percentage diameter of small and large arterioles (percentage of control) was significantly lower at a MAP of 70, 60, and 50 mm Hg in the hypothermic group than the normothermic group. Our in vivo study demonstrates that hypothermia impairs autoregulatory vasodilation of pial arterioles in response to hemorrhagic hypotension. IMPLICATIONS: Deliberate mild hypothermia has been proposed as a means of providing cerebral protection during neurosurgical procedures. Our results suggest that cerebral blood flow autoregulation in response to hemorrhagic hypotension may be impaired during hypothermic conditions, indicating the importance of maintaining perfusion pressure during hypothermic therapy to prevent cerebral ischemia.

Animals↗

Phenylarsine oxide inhibits heat shock protein 70 induction in cultured guinea pig gastric mucosal cells.

Phenylarsine oxide (PAO) forms a stable ring complex with vicinal dithiols that can be reversed with 2,3-dimercaptopropanol (DMP) but not by dithiothreitol (DTT) or 2-mercaptoethanol (2-ME). PAO at 2 microM or higher inhibited heat shock protein 70 (HSP70) induction within minutes in cultured guinea pig gastric mucosal cells exposed to heat (43 degrees C) for 30 min. PAO did not affect the nuclear translocation and phosphorylation of heat shock factor 1 (HSF1) induced by heat stress, but it completely blocked the binding activity of HSF1 to the heat shock element (HSE), leading to the block of expression of HSP70 mRNA and accumulation of HSP70 in the cells. These inhibitions were completely reversed with 2 microM DMP but not with 0.1 mM DTT or 1 mM 2-ME, suggesting specific interactions between PAO and vicinal dithiol-containing molecules. Thioredoxin (Trx) reversed the inhibition of the binding activity of HSF1 in whole cell extracts prepared from PAO-treated, heat-stressed cells. Our results suggest that PAO may react with vicinal-containing molecules including Trx and specifically block the interaction between HSF1 and HSE.

Animals↗

Regulation of growth and apoptosis of cultured guinea pig gastric mucosal cells by mitogenic oxidase 1.

We previously reported that primary cultures of guinea pig gastric pit cells expressed all of the phagocyte NADPH oxidase components (gp91-, p22-, p67-, p47-, and p40-phox) and could spontaneously release superoxide anion (O(2)(-)). We demonstrate here that pit cells express a nonphagocyte-specific gp91-phox homolog (Mox1) but not gp91-phox. Inclusion of catalase significantly inhibited [(3)H]thymidine uptake during the initial 2 days of culture. Pit cells, matured on day 2, slowly underwent spontaneous apoptosis. Scavenging O(2)(-) and related oxidants by superoxide dismutase plus catalase or N-acetyl cysteine (NAC) and inhibiting Mox1 oxidase by diphenylene iodonium activated caspase 3-like proteases and markedly enhanced chromatin condensation and DNA fragmentation. This accelerated apoptosis was completely blocked by a caspase inhibitor, z-Val-Ala-Asp-CH(2)F. Mox1-derived reactive oxygen intermediates constitutively activated nuclear factor-kappaB, and inhibition of this activity by nuclear factor-kappaB decoy oligodeoxynucleotide accelerated their spontaneous apoptosis. These results suggest that O(2)(-) produced by the pit cell Mox1 oxidase may play a crucial role in the regulation of their spontaneous apoptosis as well as cell proliferation.

Animals↗

Rapid purification of human DNase I using mouse monoclonal anti-DNase I antibodies and characterization of the antibodies.

Five anti-human deoxyribonuclease I (DNase I) monoclonal antibodies were obtained from BALB/c mice immunized with DNase I purified from human urine. Four of them inhibited DNase I enzyme activity, as did a rabbit polyclonal antibody; these 4 did not have immunostaining ability. The remaining one had immunostaining ability but no inhibitory activity. A Sepharose 4B column conjugated with 1 of the 4 antibodies that had inhibitory activity effectively adsorbed and eluted the DNase I enzyme; this did not occur with the rabbit polyclonal antibody. We showed that adding an immunoaffinity chromatography step made the purification of human DNase I easier and faster than the conventional procedure.

Animals↗

The characteristics and applications of recombinant cholesterol dehydrogenase.

Mass production of an r-CDH derived from Nocardia species was made possible by gene technology. (Horinouchi et al., Applied and Environmental Microbiology, 57, 1386-1393 (1991)). However, the characteristics of the r-CDH have not been studied in detail and have not been improved enough for industrial use. We accordingly characterized both the native-CDH and the r-CDH prepared from Streptomyces lividans. Both CDHs were monomers with molecular masses of 37 kDa. The Km of r-CDH was 2.50 x 10(-3) M for cholesterol and 2.33 x 10(-4) M for NAD. The activators of CDHs were TritonX-100 and cholate. TritonX-405, Ag+, and Zn2+ inhibited both enzymes. The residual activity of native CDH after heat treatment was 32% (37 degrees C, 60 min), while the r-CDH showed a residual activity of 87% (37 degrees C, 60 min). The r-CDH is an enzyme with high substrate specificity for cholesterol as well as native CDH and higher thermal stability than native CDH. We have developed a novel serum cholesterol assay using the r-CDH, which permits the direct measurement of cholesterol by measuring NADH reaction products. We conclude that this r-CDH enzyme is useful and can be used to measure cholesterol in a clinical chemistry setting.

Cholesterol↗