Biomedical subjects
Publications and source records attributed to S Oshida.
An improved high resolution single method for orosomucoid ORM1 and ORM2 phenotyping using isoelectric focusing (IEF) in wide-scales ultrathin layer polyacrylamide gels of pH range 4.2 to 4.9 is presented. The method is reliable, simple, and provides an alternative for the three currently required ones for typing this genetic system.
Isoelectric focusing was used to investigate the genetic variants of the human plasma orosomucoid ORM1 and ORM2 gene loci in samples of Egyptians, Sudanese, and Qataris. The study populations were classified into 28 ORM phenotypes determined by 10 ORM1 and 9 ORM2 alleles that included 2 new alleles, designated ORM1*B13 and ORM2*H21. Family studies of these new alleles are in accordance with codominant autosomal inheritance. A new interpretation for two previously reported alleles, ORM1*C6 and ORM2*H17, is also presented.
An improved method for DNA polymorphism typing of D1S80 VNTR locus and its application to paternity testing are described. For accurate estimation of the length of polymorphic DNA fragments, the size marker was labeled with fluorescence different from that of PCR primers, and co-electrophoresed as an internal standard. The dualcolour system of fluorescence image analyzer was used to detect the fragments and determine their size. This internal marker method could successfully overcome the problems of band pattern distortion and tailing, besides it allows easy and accurate interpretation of the DNA types. Our results indicate that the internal marker method is much more accurate than the method of using size marker in gel, even with the presence of distortion or tailing of the band patterns. Family studies applying this method showed complete agreement between the observed and predicted types.
Male rats of the Wistar strain were divided 4 groups, and give a liquid diet of control feed, bezafibrate (150 mg/kg), ethanol, and ethanol plus bezafibrate for 5 week. The effect of bezafibrate supplementation on rats fed ethanol was examined in terms of the fatty acid composition of the phospholipids in the erythrocyte membrane. In the phospholipids profiles of erythrocyte membranes, PI was significantly decreased. The decrease in PI caused by bezafibrate appeared to substantially affect the membrane and consequently lead to changes in the membrane anchor. In the fatty acid composition of the PC, C20: 4 was significantly decreased in the group receiving alcohol (p < 0.05) but increased in the groups receiving bezafibrate (p < 0.05). In the fatty acid composition of the PE, C16: 0 was significantly increased in the three groups when compared with the control, and C20: 4 was decreased in the alcohol group (p < 0.05). In the fatty acid of SM and PI, C20: 4 was decreased and C18: 0 increased in the alcohol group. In the PS, C14: 0 was increased in the alcohol group, and decreased in the alcohol plus bezafibrate group (p < 0.01). The levels of arachidonic acid in the total fatty acids that constituted the membrane phospholipids were decreased in the rats given ethanol (p < 0.05). However, arachidonic acid in the group of bezafibrate supplementation on rats fed ethanol were elevated in comparison with the alcohol group (p < 0.05). With decreasing arachidonic acid as a marker of alcohol tissue injury following chronic alcohol intake, the effects of bezafibrate supplementation appear to contribute to membrane fluidity by altering the biochemical flexibility of the membrane.
Allele frequency distributions for the D1S80 (MCT118) and HLA DQ alpha loci were determined in a Chinese population sample using the polymerase chain reaction (PCR). A total of 25 alleles and 100 phenotypes were observed for D1S80. The frequency of allele 18 was higher than allele 24 only in this Chinese population when compared to other reported populations. A total of 6 alleles and 21 possible phenotypes were observed for HLA DQ alpha. The power of discrimination was 0.97 and 0.93 for D1S80 and HLA DQ alpha, respectively.
Monitoring of the arterial oxygen saturation (SaO2) in two sleeping infants was reported. They were a couple of female twins and had grown up normally after admission to NICU (Neonatal Intensive Care Unit) for a month. We investigated their SaO2 with the puls-oximeter at the terms between the 2nd and the 10th month. There was no significant difference between SaO2 in prone positions (n = 29) and that in supine positions (n = 30). In the infant with the sniffing conditions, however, SaO2 in prone positions seemed to be slightly lower than that in the healthy conditions. The slightly low SaO2 would give no effects on the healthy infants, but it may cause ALTE (Apparent Life Threatening Event) or death from SIDS (Sudden Infant Death Syndrome) to some infants who are not in good health and/or who are prone to get into ALTEA or SIDS. Especially, in the sniffing condition with hyperthermia, as the oxygen dissociation curve sifts to right and SaO2 degradation appears easily, sudden infant death may be possible to happen in prone position.
We attempted to analyze biological specimens simultaneously for alcohols and hydrogen cyanide. A headspace gas chromatographic method with thick film wide bore column (PEG 20M) for the simultaneous determinations of methanol, ethanol, n-propanol and hydrogen cyanide in blood has been developed. This method was applied for the determinations of methanol, ethanol and hydrogen cyanide in a forensic autopsy case and animal experiments.
Five patients who presented to an emergency room and did not have other injury and disease with acute alcohol intoxication were analyzed about blood and urine ethanol, acetaldehyde, acetate and acetone levels. The average concentrations of ethanol, acetaldehyde, acetate and acetone in blood were 37.0 mM (1.7 mg/ml), 18 microM, 1.00 mM and 18 microM, respectively and the concentrations in urine were 50.8 mM (2.3 mg/ml), 37 microM, 0.79 mM and 47 microM, respectively. Clinical symptoms were concerned with both ethanol concentration and concentrations of ethanol metabolites. Their symptoms of acute alcohol-intoxicated patients were caused by the ethanol concentrations which was less than the levels reported in early studies.
In May, 1993, our committee of the Medico-Legal society of Japan published an article entitled "Recommendations for Preventing Transmission of Infection with Biohazard in the Practice of Legal Medicine" in The Japanese Journal of Legal Medicine. Concerning this article, we received a lot of useful comments and opinions from many members of the society, for which we are very grateful. We also received a lot of requests for further information concerning details on the equipment and its practical applications. Since a lot of departments of Legal Medicine at several universities throughout Japan at present are remodelling their facilities in order to be able to perform autopsies, with respect to such circumstances, our committee felt the need to provide information on basic conditions concerning the prevention of transmission of infections with hazards.
In Japan recently there arose a sudden need for special guidelines concerning medicolegal autopsy and the handling of medicolegal materials. In this respect, the Medico-legal Society of Japan, Committee on Expert Opinions has produced a guideline, entitled "Recommendations for Preventing Transmission of Infection with Biohazard in the Practice of Legal Medicine", which was then introduced to all members in Legal Medicine. This guideline contains First: Explanations about the most important types of Biohazard diseases, the dangers and ways of transmission. Second: Explanations concerning preventive measures in medicolegal autopsy, for handling of medicolegal materials and investigation samples, also concerning the equipment and tools within the autopsy room, as well as measures for correct disinfection. These guidelines, form the base on each department of the Medico-Legal Society of Japan, are asked to adjust their own practical manual in order to improve the safety of autopsy conditions.
Semen was obtained from 45 healthy males, and its plasma proteins of low side molecular weight obtained by filtration with a Molcut II (Millipore) were analyzed by means of capillary electrophoresis (CE) for an application to forensic medicine, with a phosphoric acid of pH 4.5 and a column of 100 cm x 75 microns ID. There were noted 27 to 36 peaks on the chromatograms, two sections of which showed individual differences in the chromatographic patterns. The chromatographic patterns between 20.0 and 22.0 minutes were classified into 2 groups: I (66.7%) and II (33.3%). The main peak appearing at about 36 minutes was divided between two patterns: A (53.3%) and B (46.7%). The serum, saliva, and vaginal fluid were also analyzed for low-side-molecular-weight proteins by means of capillary electrophoresis, but their levels contained low and their chromatographic patterns different from those of seminal plasma. The patterns of the whole seminal plasma, saliva and serum with pH 7.0 phosphate buffer were different and appeared to be applicable for identifying such samples.
n-Pentane and isopentane have a wide range of use, for example, for cleaning precision machinery, extracting essence and oil, and as liquid fuel for now very popular disposable lighters. They are contained in liquefied petroleum gas and natural gas as trace constituents. In our present experiments, we studied the metabolism and toxicity of these n-pentane and isopentane metabolites. Male mice of ICR strain were exposed to about 5% n-pentane for one hour while the oxygen in the environmental air was maintained at about 20%. Then their blood and liver tissue were collected and analyzed by means of GC and GC-MS. The metabolites thus obtained were 2-pentanol, 3-pentanol and 2-pentanone. The same procedure was repeated with isopentane; 3-methyl-2-butanol, 2-methyl-2-butanol and 3-methyl-2-butanone were detected as the resultant metabolites. In the presence of the NADPH-generating system liver microsomes were made to react to the substrate of saturated n-pentane or isopentane aqueous solution at 37 degrees C for one hour. As a result, the same metabolites were produced as obtained in the exposure experiment. It was therefore suggested that n-pentane and isopentane were metabolized chiefly by liver microsomes. Male mice of ICR strain were fed with 80 mg/kg b.w. of phenobarbital for consecutive four days and exposed to n-pentane or isopentane for one hour. This resulted in an increase in the amount of 2-pentanol and 2-pentanone in the n-pentane inhalation and 2-methyl-2-butanol in the isopentane inhalation experiment. The toxicity of each metabolite was studied on cultured cells. The metabolites were individually mixed with HeLa S3 cell suspension, incubated for three days, and their concentration which inhibited the growth of cells by 50% (IGC 50) were compared. It was demonstrated as a result that the IGC 50 for any of the metabolites was lower than that for methanol, ethanol or acetone used as control.
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Carbon monoxide (CO), total hemoglobin (Hb) and carboxyhemoglobin (HbCO) in the blood and reddish discolored body cavity fluids of cadavers which had not been exposed to fire and CO were analyzed. In 13 cadavers found on land, the maximum saturation of HbCO in the blood was 3.6%, and was 10.1% in the body cavity fluids. There was only one case in which the HbCO saturations in the body cavity fluids were more than 10%. In seven drowned bodies found in fresh water, the highest HbCO saturation in the blood was 6.1%, and was 44.1% in the body cavity fluids. There were three cases in which the HbCO saturations in the body cavity fluids were more than 10%. In 12 drowned bodies found in sea water, the HbCO saturations in the blood were not more than 6.2%, and the maximum saturation of HbCO in the body cavity fluids was 83.7%. There were eight cases in which the HbCO saturations in the body cavity fluids were more than 10%. The results seem to indicate that the interpretation of HbCO saturation in the blood would not be affected significantly by the postmortem formation of CO, and that body cavity fluids should not be used for CO determination.
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There were 1,985 fatal cases of CO poisoning in the Tohoku district of Japan in the period from 1969 to 1980. Among them, 1,322 cases were suicidal, 662 accidental, and one homicidal, in which a man killed his wife with the self-made CO gas to obtain by fraud a large amount of life insurance. Our nationwide survey revealed four other cases of homicidal CO poisoning and two cases of camouflaged CO poisoning. The police and police surgeons should be cautious enough in cases of CO poisoning.