Biomedical subjects

K Urano

Publications and source records attributed to K Urano.

At least 19 recordsLinked to original sources

PubMed2001

Method for measuring mutagen formation potential (MFP) on chlorination as a new water quality index.

A novel water quality index, the mutagen formation potential (MFP) is proposed for use in evaluation of the quality of drinking water which may contain pollutants capable of forming mutagens when chlorinated under the conditions used in water purification processes. A method for measuring MFP was established as follows. The water sample to be tested is diluted until the TOC reaches 3-4 mg l-1, the pH is adjusted to 7.0 +/- 0.2, sodium hypochlorite is added to obtain conditions where Cl/TOC = 3-4 mg Cl (mg C)-1, and the water sample is left standing for 24 +/- 2 h at room temperature. Thereafter, 21 of the chlorinated water sample at pH 2.0 +/- 0.1 is passed through a Sep-Pak Plus CSP-800 cartridge to adsorb any mutagens formed, and DMSO is applied to the cartridge to desorb the mutagens. Then, a 2 ml sample of the eluate is collected after the DMSO had begun to flow out of the cartridge and evaluated by the Ames Salmonella mutagenicity assay (preincubation method).

Chlorine Compounds↗

PubMed2001

Convenient substitute indices to toxic equivalent quantity for controlling and monitoring dioxins in stack gas from waste incineration facilities.

It is necessary to develop technical systems for controlling and monitoring dioxins in stack gas from waste incineration facilities. However, it required much labor, high technical skill and extreme cost to measure dioxins for obtaining the toxic equivalent quantity (TEQ) values. In this study, the positive correlations of the WHO-TEQ values of PCDDs/PCDFs/Co-PCBs or PCDDs/PCDFs with the conventional I-TEQ values of PCDDs/PCDFs were confirmed beforehand for a large number of incineration facilities in a wide concentration range of a hundred thousand times. The relationships between the TEQ values and the concentrations of total PCDDs/PCDFs, each toxic isomer of PCDDs/PCDFs, total Cl4-6BZs, or each homologue of Cl4-6BZs were investigated for various incineration facilities. And it was found that positive correlations of the TEQ values with the concentrations of total PCDDs/PCDFs, 2,3,4,7,8-P5CDF, or Cl5BZ for a large number of different incineration facilities and in a wide concentration range. Consequently, the concentrations of total PCDDs/PCDFs, 2,3,4,7,8-P5CDF and Cl5BZ, whose measuring methods are much easier than that of obtaining the TEQ values, could be used as convenient substitute indices to the TEQ values for controlling and monitoring dioxins in stack gas from various waste incineration facilities.

Air Pollution↗

PubMed2001

A measuring method of chlorobenzenes as a convenient substitute index of dioxins in stack gas from waste incineration facilities.

Much labor, high technical skill and extreme cost are required to measure dioxins for obtaining the toxic equivalent quantity (TEQ) values. On one side, it was found that there were high amounts of chlorobenzenes (CBZs) in the flue gas from waste incineration facilities, and they are also thought to be one of the principal precursors for dioxins. The concentrations of CBZs could be used as a convenient substitute index to the TEQ values for controlling and monitoring dioxins in stack gas from waste incineration facilities as shown in another report. In this study, collection efficiencies in the sampling performed by a convenient train which was composed of two bottles containing water and diethylene glycol in a cooling box, recoveries in the evaporation of the extract by an evaporator, proper procedure of the concentration by nitrogen purge were investigated for measuring CBZs. It was found that tetra-, penta-, and hexa-chlorobenzenes (Cl4-6BZs) whose boiling points are higher than ca. 240 degrees C could be sufficiently collected and recovered in the sampling and the evaporation. For sufficient recoveries of Cl4-6BZs in the concentration by the nitrogen purge, the final volume should be larger than 200 microliters with a proper receiver of the K-D concentrator at 35 degrees C. However, Cl1-3BZs were lost in each of the procedures.

Air Pollution↗

PubMed2001

Histologic study of the regeneration process of human hair follicles grafted onto SCID mice after bulb amputation.

This study examines histologically the degeneration and subsequent regeneration processes of human hair follicles whose bulb is severely damaged. Human scalp hair follicles were isolated and grafted onto immunodeficient mice after their bulb was amputated. On day 14, thickening and corrugation of the vitreous membrane, apoptosis of follicular keratinocytes, and regression of the lower portion of the follicles were observed. By day 20, mesenchymal cells had accumulated around the lower end of the follicles. From day 14 through 50, the follicular regression and apoptosis continued, and between days 30 and 40 the follicles became maximally shortened, and the vitreous membrane disappeared. By day 50 the lower end of the follicles had become cup-shaped, and the cup surrounded an aggregate of mesenchymal cells that corresponded to the dermal papilla. By day 60, all the grafted follicles had developed into anagen VI follicles, and the apoptosis had ceased. These results indicate that human scalp hair follicles whose bulb is completely destroyed enter into dystrophic telogen after restoration of the dermal papilla, then into anagen, and that the duration of the dystrophic telogen is shorter than that of the normal hair cycle.

Animals↗

PubMed2001

Induction of drug metabolism-related enzymes by methylcholanthrene and phenobarbital in transgenic mice carrying human prototype c-Ha-ras gene and their wild type littermates.

Transgenic mice hemizygously carrying human c-Ha-ras proto-oncogene, Tg-rasH2 show very sensitive and facilitated carcinogenicity to various carcinogens. In this study, activities of certain enzymes related to drug metabolism and energy metabolism were measured in microsome and cytosol fractions of livers of Tg-rasH2 mice and their wild type littermates with both sexes treated with 3-methylcholanthrene (MC) and phenobarbital (PB). Aminopyrine N-demethylase activities increased significantly in livers of all mice treated with PB. MC and PB treatments induced significant increases in activities of UDP-glucuronosyltransferase and S-adenosyl homocysteinase compared to those in the non-treated groups in microsome fractions from all mice. In cytosol fractions of livers of all mice, glutathione S-transferase activity was significantly induced in the PB treated groups. There were no significant differences in activities of lactate dehydrogenase, glucose 6-phosphate dehydrogenase, pyruvate kinase and glucose 6-phosphatase related to energy metabolism in livers and kidneys among all mice. Tg-rasH2 mice showed stable activities of enzymes related to drug detoxication and energy metabolism similar to those of non-transgenic mice. These results suggest that the human c-Ha-ras transgene may not affect drug metabolism-related enzymes, and the facilitated carcinogenic response in the Tg-rasH2 mouse is not due to these enzymatic disorders.

Adenosylhomocysteinase↗

PubMed2001

Tsutsugamushi disease in Kanagawa Prefecture, Japan: clinical report of two cases and survey of other incidences in 1998.

Tsutsugamushi disease is characterized by the early appearance of a black crust at the bitten area and the subsequent development of macular or macropapular rush on the whole body with high fever. While treatment with tetracycline derivatives and chloramphenicols is effective, delayed diagnosis or inappropriate treatment will lead to fatality. In this report, we describe two typical cases of tsutsugamushi disease and discuss other incidences in Kanagawa Prefecture, Japan, in 1998. One of the present two patients was diagnosed to be a case of the new type by Kawasaki strain of Rickettsia tsutsugamushi, while responsible strain was not identified for the other case. Since the disease is spreading widely even to suburban areas, we emphasize the need to consider the possible diagnosis of tsutsugamushi disease in patients with generalized eruption and high fever.

Anti-Bacterial Agents↗

PubMed2000

Adsorption equilibriums of principal herbicides on paddy soils in Japan.

Herbicides used in paddy fields during the flooding season can easily cause pollution by run-off into rivers or by other routes. It is very important to know the adsorption characteristics that influence their fate in the soil. The adsorption equilibriums have often been expressed by Henry equations, and the values of equilibrium constant, Kd, are estimated from the adsorption constants, K(OC), based on organic carbon contents of soils. There is little information concerning the equilibrium values expressed by the Freundlich equations, and insufficient information on the actual concentration levels in the paddy field. Therefore, adsorption equilibriums of the five principal herbicides: esprocarb, mefenacet, pretilachlor, simetryn and thiobencarb, on five kinds of paddy soil in Japan were investigated. It was found that their equilibrium values were better expressed by the Freundlich equation for concentration levels for the paddy fields, and that the values for the adsorption coefficient, n, varied from 1.0 to 1.6. Values for the coefficient, k, were in the range of 29-420 mg(1 - 1/n) l(1/n)/kg-dry, and the values were poorly related to solubilities in water or to the octanol-water partition coefficients of the herbicides. For each herbicide, except for simetryn, the values of k among the soils differed by 2-3 times, and no correlation could be found with the organic carbon contents, specific surface areas, pH, cation exchange capacity or major minerals of the soils. The adsorption equilibriums calculated from the values of adsorption constant Kd by the values of K(OC) in the literature were found to be very different from the experimental equilibriums. From the experimental values of coefficient k and n of the Freundlich equation, the maximum runoff concentrations of the herbicides were preliminarily estimated by a simple equilibrium model.

Absorption↗

PubMed2000

Histologic and cell kinetic studies of hair loss and subsequent recovery process of human scalp hair follicles grafted onto severe combined immunodeficient mice.

To establish a model for studying human scalp hair, individually isolated hair follicles were grafted onto back skin of severe combined immunodeficient mice. Histologic changes and cell kinetics in the hair loss and subsequent recovery process were investigated. In the dystrophic stage (from day 7 to 30), all the hair shafts became dystrophic and were shed. Thickening and corrugation of vitreous membrane, apoptosis, and regression of the lower part were observed in the grafted hair follicles. 5-bromo-2'-deoxy-uridine-labeled cells were not detected in the lower end of the follicles, and keratin 19-positive cells appeared there. At the end of this stage their lower part was maximally retracted, secondary germ remained beneath the bulge, and the vitreous membrane disappeared. In the regeneration stage (from day 30 to 50), the same histologic findings as those at the end of the dystrophic stage were observed. The keratin 19-positive cells in the secondary germ, however, were replaced with keratin 19-negative and 5-bromo-2'-deoxy-uridine-labeled cells. Then, differentiation into an inner root sheath and a hair shaft began, and apoptosis was terminated. In the stable growth stage (from day 40 to at least 150), the grafted follicles were immunohistochemically and light microscopically identical with the normal anagen hair follicles except for the presence of melanin incontinence. These findings suggest that the grafted hair follicles entered into dystrophic catagen, subsequently dystrophic telogen, then returned to normal anagen follicles, and that stem cells or their close progeny in the secondary germ play an important part in the recovery process.

Animals↗

PubMed2000

Decontamination of human xenotransplantable tumor with mouse hepatitis virus by implantation in nude rat: a case report.

A human tumor xenograft contaminated with mouse hepatitis virus (MHV) was implanted in a nude rat in order to decontaminate the tumor line. The decontamination failed in the first trial, but succeeded in the second trial. The difference between the two trials was the duration of implantation of the tumor in the nude rat, i.e., 12 days in the first and 24 days in the second trial. Duration of implantation might be a factor in the decontamination of transplantable tumors infected with MHV by passaging in the nude rat.

Animals↗

PubMed1999

The quantitative and qualitative defect of CD4+ CD45RO+ memory-type T cells are involved in the abnormality of TH1 immunity in atopic dermatitis patients.

Peripheral blood mononuclear cells (PBMCs) obtained from atopic dermatitis (AD) patients produced low levels of IFN-gamma in response to Dermatophagoides farinae antigen (Der f Ag) plus IL-2 or OKT3 MoAb in contrast with PBMCs obtained from healthy donors. The reduced IFN-gamma production in AD patients' T cells appeared to be derived from the defect of CD4+ T cells but not CD8+ T cells. Indeed, from the cytoplasmic staining analysis of cytokines, it was demonstrated that the frequency of IFN-gamma producing CD4+ T cells (TH1 cells) in AD patients was markedly lower than that of healthy donors. From the phenotypic analysis using flow cytometry, it was also found that the number of CD4+ CD45RO+ memory type T cells was significantly reduced in AD patients compared with that of healthy donors. In addition to quantitative defect of memory type CD4+ T cells, functional defect of CD4+ CD45RO+ memory type T cells was also demonstrated in AD patients. Enriched CD4+ CD45RO+ T cells obtained from AD patients, who exhibited greatly reduced delayed-type hypersensitivity (DTH) response in tuberculin test, showed no significant TH1 immunity in terms of IFN-gamma production by stimulation with OKT3 MoAb or purified protein derivative (PPD). Thus, the immunological abnormality of TH1 immunity in AD patients appeared to be induced in concomitant with both the quantitative and qualitative defect of memory type CD4+ T cells.

Antibodies, Monoclonal↗

PubMed1998

Validation of transgenic mice harboring the human prototype c-Ha-ras gene as a bioassay model for rapid carcinogenicity testing.

Studies were conducted to validate the transgenic (Tg) mice harboring human prototype c-Ha-ras gene, namely the rasH2 mice (CB6F1), as a model for rapid carcinogenicity testing. Short-term (26 weeks) carcinogenicity testing of 18 mutagenic (Salmonella) trans-species carcinogens, two mutagenic single-species (mouse-only) carcinogens, six non-mutagenic trans-species carcinogens, one non-mutagenic single-species (mouse-only) carcinogen, four mutagenic non-carcinogens and four non-mutagenic non-carcinogens were completed. The studies revealed that the Tg mice are able to detect various types of mutagenic carcinogens and may also detect various non-mutagenic carcinogens within 26 weeks. Dose-dependent tumor responses were observed with various carcinogens except for a few equivocal cases. The validation studies also revealed that the Tg mice are generally much more susceptible to both mutagenic and non-mutagenic carcinogens than control non-Tg mice. Most of the malignant tumors were observed in the carcinogen-treated Tg mice and only very few or none in the corresponding non-Tg mice. Most of the carcinogens tested induced some of the target organ tumors observed in B6C3F1 mice in a 2-year bioassay as well as certain types of tumors specific to the Tg mice, i.e. lung alveolar epithelial tumors, spleen hemangiosarcomas, forestomach squamous cell tumors. No significant tumor induction has been observed in the Tg mice either with mutagenic or non-mutagenic non-carcinogens. Although further validation studies are still required, the rasH2 mouse seems to be a promising candidate as an animal model for the development of a rapid carcinogenicity testing system.

Animals↗

PubMed1998

Statistical procedures for estimating the detection limit and determination limit of the Ames Salmonella mutagenicity assay.

Novel and flexible procedures for estimating the detection limit as well as the determination limit of the Ames mutagenicity assay were proposed to evaluate the genetoxicity of a water sample. The accumulated data under the test conditions of TA 100-S9 by our group were taken as examples and analyzed to estimate the detection limit and the determination limit. The detection limit was estimated at 1.7 as the MR value when duplicate plates were used in the negative control test. However, it decreased to 1.4 as the MR level when quadruple plates were used in the negative control test. Therefore it was found that the sensitivity of the Ames mutagenicity assay was improved very easily by increasing the number of plates for the negative control test from two to four. The application of the conventional twofold rule to the data obtained with the strain TA100 was considered too conservative. The determination limit was regarded at 2.2 as the MR value under the following conditions: (a) quadruple plates were used in the negative control test; (b) three dose-steps including negative control step were designed at regular intervals; and (c) duplicate plates were used for each dose-step. It was proved by comparing data of two students that the detection limit and the determination limit estimated in this study were considered acceptable to any well trained students.

Biometry↗

PubMed1998

Mutagenic activities of exhaust gas and ash from sludge incineration plants.

We have found that the exhaust gas from many incineration plants of municipal solid wastes (MSW) show significant mutagenic activities. The mutagenic activities of exhaust gas from incineration plants of the other wastes have not been studied in detail. Here, we analyzed the mutagenic activities and compounds in exhaust gas and ash from seven sludge incineration plants. Some samples of the exhaust gas from the sludge incineration plants showed high mutagenic activities; although, none of the ash residues showed mutagenic activities. There was no relationship between the mutagenic activities and furnace types, the plant size or the apparent residence time of the gas in the furnace. The mutagenic activities of the exhaust gas were produced during incomplete combustion at lower temperatures. Direct mutagenic activities without S9 mix were higher than indirect mutagenic activities with S9 mix which was made from rat liver homogenate and was used to test mutagenic activity after metabolism. These results are different to those of MSW incineration plants. We analyzed the mutagenic compounds in the exhaust gas by GC/MS after fractionation by HPLC, but they could not be identified. We found that the mutagenic compounds in the exhaust gas were different from the compounds that were produced from the MSW incineration plants. We believed that these mutagenic compounds might be non-volatile and more polar than the heterocyclic amines, Trp-P-2, Trp-P-1 and Glu-P-1, which are typical mutagenic compounds in sewage and sludge.

Animals↗

PubMed1998

Validation of transgenic mice carrying the human prototype c-Ha-ras gene as a bioassay model for rapid carcinogenicity testing.

Carcinogenicity testing is indispensable for identifying environmental carcinogens and for evaluating the safety of drugs in the process of development. Conventional 2-year rodent bioassays are one of the most resource-consuming tests in terms of animals, time, and costs. Development of rapid carcinogenicity testing systems that can assess carcinogenicity within a short period has become a social demand and is essential to improve efficacy in the identification of environmental carcinogens as well as in the development of new drugs. In this review we introduce the rapid carcinogenicity testing system using transgenic (Tg) mice carrying the human prototype c-Ha-ras gene, namely rasH2 mouse (CB6F1-TgHras2 mouse is the same mouse). The studies have been conducted to validate the rasH2 mouse as a model for the rapid carcinogenicity testing system. Our current validation studies revealed that rasH2 mice are able to detect various types of mutagenic carcinogens within 6 months. The rasH2 mice may also be able to detect various nonmutagenic carcinogens. The validation studies also revealed that rasH2 mice are generally much more susceptible to both mutagenic and nonmutagenic carcinogens than control non-Tg mice. No significant tumor induction has been observed in rasH2 mice with either mutagenic or nonmutagenic noncarcinogens. More rapid onset and higher incidence of more malignant tumors can be expected with a high probability after treatment with various carcinogens in the rasH2 mice than in control non-Tg mice. The rasH2 mouse appears to be a promising candidate as an animal model for development of a rapid carcinogenicity testing system.

Animals↗

PubMed1997

New immunodeficient (nude-scid, beige-scid) mice as excellent recipients of human skin grafts containing intraepidermal neoplasms.

Engraftment of normal or lesional human skin onto nude or SCID (severe combined immunodeficiency) mice has been used as an in vivo experimental model. However, this model has some limitations, such as shrinkage and loss of the grafted skin over time. To improve the experimental model, we have produced two new SCID-lineage mouse strains, BALB/cA-nude-scid (nu/nu, scid/scid) and BALB/cA-beige-scid (bg/bg, scid/scid) mice, by the method of cross intercross. Intraepidermal neoplastic lesions such as Bowen's disease were grafted onto the back of the mice of these strains. The rate of reduction in the size of the grafts was lower on nude-scid and beige-scid mice than on SCID mice. Rates of survival of neoplastic cells in the grafts were higher in nude-scid mice than in SCID and beige-scid mice (SCID mice 38%, nude-scid mice 55%, beige-scid mice 38%). Neoplastic cells of Bowen's disease grafted onto a beige-scid mouse proliferated and invaded the dermis during 233 days of observation, confirming the progression to invasive squamous cell carcinoma from carcinoma in situ. The present study revealed that nude-scid and beige-scide mice newly produced by us provide a very useful in vivo experimental model for the investigation of carcinogenesis and tumor progression in human skin.

Animals↗

PubMed1997

Kinetic characterization of binding and internalization of fractionated [3H]heparin in rat liver parenchymal cells in primary culture.

The binding and internalization of fractionated [3H]heparin (FH) was kinetically analyzed in rat liver parenchymal cells to clarify its cellular uptake mechanism. The binding of FH to the cell surface was saturable with the dissociation constant (Kd) of 53.5 nM and a maximum binding capacity (Bmax) of 19.9 pmol/mg protein. The binding of FH to the cell surface was competitively inhibited not only by heparan sulfate, a polyanion analogous to heparin, but also by rose bengal, an organic anion, suggesting the binding is based on an electric interaction requiring an anionic charge for substrates and consistent with the earlier suggestion of the involvement of the scavenger-like receptor. According to kinetic model analysis, the rate constants of association (K(on)), dissociation (k(off)), and internalization (k(int).app) were estimated to be 0.0005 nM-1 min-1, 0.0112 min-1, and 0.0056 min-1, respectively. Although both Kd and Bmax were larger than those reported in Kupffer cells, suggesting lower affinity and higher capacity in liver parenchymal cells, the apparent internalization rate constant was similar to that in Kupffer cells. We thus provided additional evidence suggesting that a scavenger-like receptor exists in rat liver parenchymal cells, and then kinetically characterized the surface binding and internalization of fractionated heparin by this receptor.

Animals↗

PubMed1997

PUVA treatment of human keratinocytes suppresses the surface expression of intercellular adhesion molecule 1 (ICAM-1) and inhibits adherence of PHA-blasts in vitro.

To determine the basis of the therapeutic efficacy of Psoralens and UVA (PUVA) in inflammatory skin diseases, the effect of PUVA on the expression of cell adhesion molecules of keratinocytes was investigated in vitro. The addition of IFN-gamma and TNF-alpha to human keratinocytes in culture up-regulated the expression of ICAM-1 and HLA-DR on the cell surface. The cultured human keratinocytes were exposed to UVA light in the presence of 8-methoxypsoralen (PUVA). The ICAM-1 and HLA-DR surface molecules were stained by monoclonal antibodies, and the intensity of the resultant fluorescence was analyzed by FACS-can. PUVA treatment suppressed the expression of these cell surface molecules, with increasing UVA fluence. Moreover, PHA-blasts failed to adhere to PUVA treated keratinocytes. When keratinocytes were treated with PUVA prior to the addition of IFN-gamma and TNF-alpha, ICAM-1 and HLA-DR expression was suppressed. These results suggest that one of the therapeutic mechanisms of PUVA in inflammatory skin diseases is by inhibition of the adhesion of activated lymphocytes to keratinocytes due to suppression of cell surface molecules. It also suggests that PUVA may be useful as maintenance therapy for inflammatory skin diseases.

Adult↗

PubMed1996

Characteristics of lethal substances and the removal effect of molecular sulfur in the Salmonella mutagenicity test for river sediments.

The lethal action and the effect from the removal of molecular sulfur by copper treatment were investigated in the Salmonella mutagenicity test for river sediments. Survival ratios of Ames Salmonella strains for five different sediment extracts were tested, and it was found that many of the river sediments showed the lethal action on TA98 and TA100, and the lethal action was stronger in the sample from which larger amount of extracts per unit weight were obtained. It was also found that the lethal action was reduced with the addition of the metabolizer S9 mix, and TA98 was more resistant than TA100 against the lethal substances in sediment extracts. Further, removal of molecular sulfur, one of the lethal substances, by copper treatment raised survival ratios of the tester strains. On high-pressure liquid chromatographic fractionation, the lethal substances, including molecular sulfur distributed in non-polar and slightly polar fractions, and medium or high polar fractions did not reveal lethal action. Consequently, the removal procedure of molecular sulfur was proper for applying on the fraction contained non-polar and slightly polar compounds.

Japan↗