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Biomedical subjects

M Miyaji

Publications and source records attributed to M Miyaji.

At least 127 records · Page 7Linked to original sources

Rapid, extensive and reversible vacuolation of Schizosaccharomyces pombe induced by amphotericin B.

The fission yeast Schizosaccharomyces pombe has no large vacuoles under normal growth conditions, although budding yeasts usually have large central vacuoles. The minimum inhibitory concentration of amphotericin B to S. pombe was 0.5 microgram ml-1; treatment with 0.2 microgram ml-1 for 20 min induced rapid and extensive vacuolation in S. pombe exponential phase cells. Growth rate of the cells with 0.2 microgram ml-1 amphotericin B was much reduced for 6 h, showing extensive vacuolation. Vacuolation in itself was not fatal: on removal of the drug, most cells recovered gradually and eventually multiplied.

Amphotericin B↗

Ubiquinone systems of Coccidioides immitis, the causative agent of coccidioidomycosis.

The ubiquinone (coenzyme Q) systems of eleven strains of Coccidioides immitis were determined by high performance liquid chromatography (HPLC). The ubiquinone profile of the fungi was shown to be homogeneous: in all of the strains, ubiquinone-10 (Q-10) was demonstrated to be the major component, with Q-9 as a minor component. The results imply that the ubiquinone system may serve as an additional phenotypic criterion for identifying the fungus.

Coccidioides↗

The isolation of Cryptococcus neoformans from pigeon droppings and serotyping of naturally and clinically sourced isolates in China.

This is the first report on the isolation of Cryptococcus neoformans from pigeon droppings in China and their serotypes. C. neoformans colonies which produced brown colonies on caffeic acid-cornmeal agar were found in Twenty-five out of thirty-six samples of pigeon droppings. Fifty-one colonies randomly picked from the positive samples were identified as C. neoformans by a commercially available kit for carbon source assimilation test and Christensen's urea agar. Forty (78%) out of the 51 strains were serotyped as A and 11 (22%) as AD. At the same time, seventeen out of nineteen clinical isolates were serotyped as A and 2 as B. There are three findings in our results. One is that only C. neoformans var. neoformans strains could be isolated from pigeon droppings, although the variety gattii strains were found in the clinical isolates obtained in the same geographic site in China. The second is that serotype A strains were most frequently seen in natural and clinical materials in the southeast part of China, and serotype AD strains were isolated in pigeon droppings but not in clinical materials. The third is that the coexistence of serotype A and AD cells of C. neoformans strains in same samples of pigeon droppings were observed.

Animals↗

Mitochondrial DNA analysis of Exophiala moniliae.

Mitochondrial DNA (mtDNA) analysis with restriction enzymes, Hae III, Hind III and Msp I was performed in 17 Exophiala moniliae strains. The results were as follows: (1) E. moniliae could be classified into 10 types based on restriction patterns, (2) E. moniliae is suggested to be a complex organism because of extensive mtDNA polymorphism among strains like E. jeanselmei and (3) two types of E. moniliae are identical with two types of E. jeanselmei. These results suggest that E. moniliae is not genetically defined from E. jeanselmei and the taxonomical status of E. moniliae requires reevaluation.

DNA Restriction Enzymes↗

Molecular epidemiology of Cladosporium carrionii.

Mitochondrial DNA (mtDNA) diversity was investigated in 38 isolants of Cladosporium carrionii in Africa (3 isolants), Australia (10), China (15) and Venezuela (10). C. carrionii was classified into 4 mtDNA types (Type I to Type IV) based on the restriction patterns with MspI, Sau3AI and HaeIII. All the 15 strains in China and 9 of 10 strains in Venezuela showed Type I, while all the 10 strains in Australia showed Type II. These results suggested a correlation between the mtDNA types and geographic origin.

Africa↗

A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis.

A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis was carried out. The staining methods used were fluorescent staining, vital dye exclusion tests with erythrosin B and by Janus green and lactophenol cotton blue staining. Colony forming units (cfu) of the yeast form of eight P. brasiliensis isolates on brain heart infusion agar (BHIA) supplemented with 4% horse serum plus 5% P. brasiliensis cell extract (BHIA + HS + EXT) were examined for reliability of staining in determining the number of live fungal units in eight different isolates. Cfu on BHIA + HS + EXT plates showed an excellent plating efficiency over 96% in all isolates tested. The percentage of the live cells indicated by fluorescent staining (FL) or vital dye exclusion test with erythrosin B (EB) or Janus green (JG-1) was lower than that of cfu. By contrast, the percentage due to modified dye exclusion test with Janus green (JG-2) and that due to lactophenol cotton blue staining (LPCB) showed a close correration to that of cfu. Our results indicate that the modified dye exclusion test with Janus green and lactophenol cotton blue staining are useful for estimating cell viability of yeast form cells of P. brasiliensis.

Azo Compounds↗

An improved culture medium for detecting live yeast phase cells of Paracoccidioides brasiliensis.

The plating efficiency of standard mycological media such as brain heart infusion (BHI) agar is poor for Paracoccidioides brasiliensis. We prepared a water-extract of yeast phase cells of P. brasiliensis and examined it for growth-enhancing activity for the fungus. The water-extract, when added to BHI agar to a concentration of 5%, improved the plating efficiency of the medium for the fungus to some extent, but the degree of improvement was considerably varied among P. brasiliensis isolates. By contrast, when the water-extract was added in combination with horse serum (4%), the plating efficiency was highly improved (to 94-99%) for all the P. brasiliensis isolates employed. The growth-enhancing factor(s) in the water-extract was heat-stable and heating at 120 degrees C for 15 min had little, if any, effect on growth-enhancing activity.

Culture Media↗

Duodenal stromal tumor with neural differentiation.

A 49-year-old woman was admitted to our hospital for detailed evaluation of an abdominal mass. Ultrasonography and computed tomography revealed two tumors, one a submucosal lesion of the duodenal second portion and the other a left adrenal tumor. Angiography showed that the tumors were hypervascular. Both tumors were removed surgically and examined histologically using hematoxylin-eosin staining. The duodenal tumor (10 x 6 x 5 cm) was initially diagnosed as a schwannoma, but immunohistochemical studies showed that it was S-100 protein negative and neuron-specific enolase positive. Therefore, this tumor was identified as a stromal tumor with neural differentiation. The left adrenal tumor was a nonfunctioning adenoma.

Adenoma↗

Effects of dibutyryl cyclic AMP and papaverine on intrahepatocytic bile acid transport. Role of vesicle transport.

The secondary messenger cyclic AMP plays an important role in regulating biliary excretory function by stimulating the transcytotic vesicle transport system, whereas papaverine exerts an inhibitory effect on this system. We therefore investigated their effects on bile acid-induced cytotoxicity and intrahepatocytic content of bile acid in primary cultured rat hepatocytes. Simultaneous addition of 1 mM dibutyryl cyclic AMP (DBcAMP), an analogue of cAMP, with 1 mM taurochenodeoxycholic acid (TCDCA) significantly decreased the release of lactate dehydrogenase (LDH) as compared with the case with 1 mM TCDCA alone (7.1 +/- 0.13% of total versus 10.7 +/- 0.3%). In contrast, 0.1 mM papaverine approximately doubled the amount of LDH (22.0 +/- 0.6% of total versus 10.7 +/- 0.3%; P < 0.01). The intracellular content of TCDCA 180 min after the administration of 1 mM TCDCA alone was 20.8 +/- 0.7 nmol/mg protein, that after simultaneous administration of 1 mM DBcAMP, 16.2 +/- 1.0 nmol/mg protein, and that after the simultaneous administration of 0.1 mM papaverine, 38.5 +/- 1.9 nmol/mg protein. A clear correlation between the release of LDH from hepatocytes and the intracellular content of TCDCA was thus observed. When given together with 1 mM taurocholic acid (TCA) or 1 mM tauroursodeoxycholic acid (TUDCA), papaverine exerted little effect on cytotoxicity or intrahepatocytic bile acid content. When cells were bathed in a medium free of bile acid after pretreatment with 1 mM TCDCA and 1 mM DBcAMP, additional exposure to DBcAMP for 30 min significantly stimulated reduction of intracellular TCDCA content (30.2 +/- 0.4% of total versus 44.0 +/- 1.4%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Natural history and URSO dissolution therapy of gallbladder stones in the elderly].

To determine how to treat silent gallstones in the elderly, 1771 autopsied cases 65 years of age or older were studied retrospectively, and 121 cases with asymptomatic gallbladder stones were followed for over 3 years. Of these, 47 cases treated with ursodeoxycholic acid (UDC) for over 3 years were investigated to determine the efficacy of UDC therapy. In the autopsied cases, the incidence of gallbladder stones was 16%, and increased with age. The ratio of males to females was 1:1.2. Only 3.2% of people with silent gallstones developed symptoms. Autopsy studies showed that the majority of people with gallstones died of unrelated causes such as benign respiratory and circulatory diseases. Only 1.8% of patients with gallstones died of acute cholecystitis or gallbladder carcinoma. UDC treatment was effective in 55.9% of patients with radiolucent stones. The percentage of patients requiring surgery due to becoming symptomatic was much lower (6.4%) in UDC-treated patients. The visualization of the gallbladder on cholecystogram improved in cases treated with UDC. These results indicate that elderly patients with silent gallstones generally develop neither symptoms nor carcinoma. Therefore, follow-up management by ultrasonography and cholecystography can be allowed. We also emphasized that a satisfactory response was obtained with long term UDC therapy.

Administration, Oral↗

[A case of gastric cancer with multiple liver metastasis treated with low dose CDDP therapy on an outpatient basis after palliative gastrectomy].

A 59-year-old-male with advanced gastric cancer and multiple liver metastasis was treated by intravenous administration of MMC (30 mg/body) and oral administration of HCFU (400mg/body/day) initially after palliative gastrectomy. CT scan revealed that liver metastatic lesions were undetectable at three months after operation. Seven months later, administration of HCFU was discontinued because liver metastatic lesions were observed again. Intravenous administration of MMC (10mg/body)+ACR (20mg/body) was performed four times weekly, but liver metastatic lesions were increasing in size. From 15 months after operation intravenous administration of CDDP (25mg/body/week) was started on an outpatient basis. After administration of CDDP 6 times, liver metastatic lesions were diminished in size. By CT scan the maximum percent reduction of liver metastatic lesions was 97% and duration of partial response was seven months. Eight months later, liver metastatic lesions regrew. At 28 months after operation, the patient died of liver metastatic lesions. This low dose regimen of CDDP on an outpatient basis is potentially useful in the treatment of advanced gastric cancer with multiple liver metastasis.

Ambulatory Care↗

Papaverine inhibits transcytotic vesicle transport and lipid excretion into bile in isolated perfused rat liver.

Papaverine is a nonspecific smooth muscle relaxant and a phosphodiesterase inhibitor. Its effects on biliary excretion of lipids and horseradish peroxidase were investigated in a single-pass isolated perfused rat liver model. A constant infusion of papaverine (1.6 mumol/min; 40 mumol/L) significantly increased bile flow (microliters per minute per gram of liver) before (2.03 +/- 0.09 vs. 1.0 +/- 0.06) and after sodium taurocholate infusion (2.77 +/- 0.10 vs. 1.88 +/- 0.11). However, papaverine significantly and reversibly reduced biliary excretion of phospholipids and cholesterol (nanomoles per minute per gram of liver) after a 1.0 mumol/min sodium taurocholate infusion, from 7.45 +/- 0.83 and 1.42 +/- 0.15 to 1.75 +/- 0.18 and 0.39 +/- 0.06, respectively (p less than 0.01), whereas secretion of bile acids was unaffected. When a 1-min pulse of horseradish peroxidase (25 mg) was infused in isolated perfused rat liver after a continuous infusion of N6,O-2'-dibutyryladenosine 3',5'-cyclic monophosphate (0.25 mumol/min; 6.25 mumol/L), horseradish peroxidase appeared in bile in an early (4 to 6 min) and late (20 to 25 min) peak. Papaverine significantly reduced the late peak, from 1.211 +/- 0.264 to 0.498 +/- 0.107 (p less than 0.01). Papaverine had no significant effects on either cyclic AMP or cyclic GMP in the liver and bile, although it has been reported that papaverine is a phosphodiesterase inhibitor. These findings indicate that papaverine inhibits biliary excretion of lipids but not bile acids, and they suggest that papaverine has an inhibitory effect on transcytotic vesicle transport independent of an increase of cyclic nucleotides in hepatocytes.

Animals↗

Comparative studies on the postantifungal effect produced by the synergistic interaction of flucytosine and amphotericin B on Candida albicans.

A turbidometric method was used to measure Candida albicans yeast cell growth and to quantitate the postantifungal effect (PAFE) after exposure to various concentrations of flucytosine and amphotericin B, alone and in combination, for 2 hr at 30 degrees C. The drug concentrations used in the PAFE assays were determined by initial MIC and FIC (fractional inhibitory concentration) evaluations. The PAFE was calculated by the difference in time (hr) required for growth of the control and test cultures to reach the 0.5 absorbance level following removal of the drug by dilution. A synergistic PAFE was evidenced with combinations of the two drugs at concentrations below their individual MICs. Combinations of flucytosine (0.012 to 0.049 micrograms ml-1) and amphotericin B (0.195 to 0.39 micrograms ml-1) produced PAFEs ranging from 6.3 to 21.8 hr. These PAFEs persisted from 0.3 to 14.7 hr longer than those achieved when each of the two agents was assayed separately.

Amphotericin B↗

Studies on the relationship between the estrous cycle of BALB/c mice and their resistance to Paracoccidioides brasiliensis infection.

A relationship between the estrous cycle and non-specific host resistance to Paracoccidiodes brasiliensis yeast cells was examined by using both sexes of adult BALB/c mice. They were divided into 6 groups, including a male group and females at proestrus, estrus, metestrus-I, metestrus-II and diestrus. The mice received yeast cells through three different inoculation routes; intravenous, intraperitoneal and intratracheal. In all of the inoculation routes, the clearance of the yeast cells was influenced by the estrous cycle. The female mice at estrus, which might have high blood estrogen levels, showed a marked clearance of the yeast cells from the blood, peritoneal cavity and lungs. These results suggested that non-specific host resistance to the yeast cells was enhanced by estrogen. All female groups inoculated by the three routes showed higher clearance of the yeast cells than the male group.

Animals↗

Tauro-beta-muricholate preserves choleresis and prevents taurocholate-induced cholestasis in colchicine-treated rat liver.

In recent clinical and animal experimental studies, ursodeoxycholic acid (UDCA) has been noted to have marked choleretic and cytoprotective actions. To define the mechanism and determine whether such favorable influence is specific to UDCA, the choleretic action of beta-muricholic acid (beta-MCA), which has a similar chemical structure, was studied using an isolated rat-liver-perfusion system. As a result, beta-MCA and taurine-conjugated beta-MCA (T beta-MCA) stimulated bile flow accompanied by elevation of bile acid output and phospholipid output, and beta-MCA caused an elevation in biliary HCO3- concentration in normal rat livers. After colchicine treatment, taurocholic acid (TCA) administration was associated with marked cholestasis while both beta-MCA and T beta-MCA still increased bile flow under the same conditions. Furthermore, simultaneous administration of beta-MCA or, more markedly, T beta-MCA reversed the effects of TCA alone in colchicine-treated rat liver; significant preventive effects against the cholestasis could be shown. These data suggest that beta-MCA and especially T beta-MCA can support choleresis even under conditions of colchicine-dependent microtubule dysfunction. The effects of T beta-MCA on organelle lipids and their intracellular transport may differ from those of TCA, presumably because of the anticholestatic and cytoprotective effects of T beta-MCA.

Animals↗

Survival rates of some terrestrial microorganisms under simulated space conditions.

In connection with planetary quarantine, we have been studying the survival rates of nine species of terrestrial microorganisms (viruses, bacteria, yeasts, fungi, etc.) under simulated interstellar conditions. If common terrestrial microorganisms cannot survive in space even for short periods, we can greatly reduce expenditure for sterilizing space probes. The interstellar environment in the solar system has been simulated by low temperature, high vacuum (77 k, 4 x 10(-6) torr), and protons irradiation from a Van de Graaff generator. After exposure to a barrage of protons corresponding to about 250 years of irradiation in solar space, Tobacco mosaic virus, Bacillus subtilis spores, Aspergillus niger spores and Clostridiun mangenoti spores showed survival rates of 82%, 45%, 28%, and 25%, respectively. Furthermore. pathogenic Candida albicans showed 7% survival after irradiation corresponding to about 60 years in space.

Bacteria↗

Synergistic postantifungal effect of flucytosine and fluconazole on Candida albicans.

The in vitro efficacy of flucytosine and fluconazole, separately and in combination, with respect to induction of a postantifungal effect (PAFE) on Candida albicans was studied. PAFE refers to the persistent suppression of fungal cell growth following a short period of exposure to an antifungal agent. A turbidometric method was used to measure cell growth and to quantitate the PAFE following exposure of C. albicans yeast cells to different concentrations of the two agents for 2 h. The PAFE was determined by the difference in time (h) required for growth of the control and test cultures to increase to the 0.5 absorbance level following removal of the drug by dilution. Minimum (MIC) and fractional inhibitory concentration determinations were made and the data used for selecting the concentrations used in the PAFE evaluations. A synergistic interaction of the two drugs at concentrations well below their individual MICs was evidenced. Flucytosine:fluconazole ratios of 1:16-1:32 at concentrations ranging from 0.024-0.098 micrograms ml-1 and from 0.78-1.56 micrograms ml-1, with flucytosine and fluconazole, respectively, induced PAFEs which persisted for 2.5 h longer than those achieved when each of the two agents was assayed separately.

Candida albicans↗