Carrier diagnosis by RFLP analysis in a family affected with infantile hypophosphatasia: case report.
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Biomedical subjects
Publications and source records attributed to A Akane.
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We developed a head-space method for the determination of blood cyanide by gas chromatography with electron-capture detection. In this technique, a reaction precolumn packed with chloramine-T was used for the conversion of hydrogen cyanide into cyanogen chloride. Since the reaction precolumn eliminated the necessity of trapping hydrogen cyanide from biological samples, blood cyanide could be analyzed quickly by acidification only. Using this method, blood cyanide levels of fire victims were determined at autopsy. The serum values of cyanide ranged from 0.11 micrograms/ml to 18.12 micrograms/ml. However, a significantly higher cyanide content was detected in the left ventricular blood than in the right. This indicates that death was caused by the fire and suggests that the collecting point of the blood sample is an important factor in the determination of inhaled cyanide. There was a positive correlation between blood cyanide and carboxyhemoglobin contents.
An analytical method of fructosamine (glycated serum protein) determination in hemolytic samples from cadavers was investigated for the postmortem diagnosis of diabetes mellitus. Fructosamine level is usually measured by the reduction of nitroblue tetrazolium (NBT) using a spectrophotometer. This assay was significantly disturbed by more than 1 g/l hemoglobin, which strongly reduced NBT by the catalytic action of the sulfhydryl and glycated groups. Glycated and total hemoglobin levels were then determined simultaneously to exclude the interferences. Total protein concentration was analyzed to eliminate dilutional effects by hemolysis. With these modifications, corrected fructosamine levels in samples containing not more than 10 g/l hemoglobin could be estimated. The assay of such hemolytic samples from 32 cadavers indicated higher fructosamine values in diabetic group than in non-diabetic subjects and the postmortem degradation of the levels as previously reported.
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The genetic markers of a Korean woman and her allegedly adopted child were compared to test her maternity. None of 21 conventional markers, including 8 red cell antigens, 6 red cell enzymes, and 7 serum proteins, excluded the maternity. These results indicate a maternal probability of 0.31 which was too low to conclude the true maternity. Human leukocyte antigen (HLA)-A, -B, -C and -DR haplotyping was also uninformative in this case. The maternity was consequently excluded by the observation of a difference in 2 of 5 variable number of tandem repeat (VNTR) markers.
We investigated the differences in the levels of carboxyhemoglobin (COHb), cyanide (HCN) and petroleum fuels (gasoline and kerosene) between left and right ventricular bloods from fire victims. COHb was slightly, and HCN and petroleum fuels were markedly higher levels in the left than those in the right. These effects were so called 'first pass phenomena' due to the circulation, diffusion and metabolization before the deaths of fire victims.
In motorcycle accidents involving two riders, medicolegal identification of the driver is necessary when one or both riders die. It is particularly important in the latter case, because the survivor almost always insists that he or she was not driving. One characteristic injury that distinguishes the driver from the passenger is inguinal contusion-laceration (accompanied internally by pelvic fracture). This injury, caused by collision of the pelvis with the fuel tank, identifies the driver.
We report on a three-occupant automobile collision in which one of the two survivors claimed that the deceased had operated the vehicle. We attempted to identify the driver. The left face and neck of the deceased appeared to have struck the vehicle's interior. The survivors were slightly injured on the right side of their faces. In Japan, the passenger side is on the left, and in this case, the right side of the vehicle had been damaged. By comparing the injuries of the deceased and the survivors in relation to the vehicle damages, we concluded that the deceased occupant had probably been a passenger, not the driver.
Two recent paternity cases are reported. In the first case of paternity exclusion, deoxyribonucleic acid (DNA) restriction fragment length polymorphisms (RFLPs) on variable number of tandem repeat (VNTR) loci with multiple alleles were informative, as well as established systems of red blood antigens, red cell enzymes, serum proteins, and human leukocyte antigens. In the second case, in which both the alleged father and the first wife were deceased, the paternal genotype was determined by using genetic markers from the second wife and four children, which then were compared with the paternal alleles of the child in question, the plaintiff in this case. The high probability of paternity (0.999,998,7) made us conclude that the man probably was the actual father. The DNA analysis by VNTR probes appears to be quite valuable in the study of paternity cases.
Two head-space gas chromatographic methods for the determination of hydrogen cyanide (HCN) in blood were demonstrated using electron capture detector (ECD). One was the double head-space gas sampling technique and the other used the reaction pre-column packed with Chloramine-T for the conversion of HCN into cyanogen chloride (CNC1). In the former method, HCN in blood was firstly released by acidification, and then the HCN gas was replaced into the another vial in which Chloramine-T solution had been placed. The head-space gas in the second vial was analyzed by gas chromatograph (GC). In the latter, HCN gas released was directly injected into the GC pre-connected the reaction column. The conversion of HCN into CNC1 was easily achieved by both methods. The use of dichloromethane as an internal standard and PTFE-faced septum for the sealing of vial greatly improved the precision and reliability of these analytical methods. The calibration curves by these methods gave good linearity and the lower detection limit taken from these plots was ca. 0.05 micrograms/ml. In particular, the reaction pre-column method, by which blood cyanide can be analyzed rapidly and accurately with only simple alteration of apparatus, is superior in the routine analysis of blood cyanide.
An unusual suicide is described in which analysis of circulating kerosene components at autopsy suggested that the victim had drunk kerosene and then had poured it over his body, ignited it, and burned to death.
The determination of petroleum fuel in the blood of burned bodies was carried out by three different gas chromatographic procedures. Seven components of gasoline (isopentane, n-pentane, 2-methylpentane, benzene, 2-methylhexane, 3-methylhexane and toluene) and five of kerosene (xylene, C9H20, mesitylene, pseudocumene and C11H24) were chosen as indicators with a coefficient of variation of 5-24%. The methods were applied to four autopsy cases with a relatively low carboxyhaemoglobin (HbCO) content. When gasoline exposure had occurred, the blood concentrations determined were almost identical whatever the components selected. Great variations in the components determined were found after kerosene exposure, and hydrocarbons greater than or equal to C14 were hardly inhaled by the victims. A higher content of fuel in the left than in the right ventricular blood observed in the autopsy cases suggests fuel inhalation just before death. The same phenomenon was also observed in the content of blood HbCO. Determinations of petroleum fuel and HbCO in both the right and left ventricular blood would be useful for the forensic diagnosis on burned bodies with a low HbCO content.
The effects of ethanol and NAD+ on the pregnenolone-to-testosterone pathway of testicular steroidogenesis in lysed Leydig cell preparations were investigated. Testosterone and four precursor steroids were determined by gas chromatography/mass spectrometry after incubation of the cell preparations with 100 microM of pregnenolone and appropriate concentrations of ethanol (1-100 mM). Concentrations of all 4-ene-steroids measured were significantly decreased even at the lowest concentration of ethanol. When NAD+ (0.1 mM) was added to the incubation medium, the levels of progesterone and 17-hydroxyprogesterone returned to control values, whereas those of androstenedione and testosterone remained decreased. These results suggest that ethanol may inhibit testicular steroidogenesis by suppressing at least two steps in the pregnenolone-to-testosterone pathway, the pregnenolone-to-progesterone step catalysed by NAD+-dependent 5-ene-3 beta-hydroxysteroid dehydrogenase/isomerase and the 17-hydroxy-progesterone-to-androstenedione step catalysed by the NAD+-independent C17-20 lyase.
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The metabolic interaction of toluene and ethanol was studied in male rabbits having received ethanol (26.0 mmol/kg PO), toluene (5.4 mmol/kg PO) or both. Compared with ethanol alone, toluene given 2 h after ethanol caused a significantly higher and more prolonged concentration of blood alcohol. A similar trend of blood alcohol was observed at the later stage with toluene given prior to ethanol. On the other hand, with simultaneous doses of the two substances, the blood toluene concentration was higher for the first 15-30 min than the ethanol control and the urinary excretion of hippuric acid, a main metabolite of toluene, was markedly decreased for the first 2 h. The blood ethanol in this group, on the contrary, was reduced until 1 h after administration. These results indicate that toluene and ethanol act reciprocally as competitive inhibitors in their metabolism after single administrations.
Characteristics of cytochrome oxidase prepared from hearts of Sprague-Dawley male rats were studied with the use of the fourth derivative spectrophotometry in respect to the cytotoxic effects of carbon monoxide (CO). CO-exposed rats tended to show lower specific activities of cytochrome oxidase than control and recovered rats. Moreover, there was a significant difference in the fourth derivative spectral features of the enzyme: CO-exposed groups indicated peaks at 412 nm in the spectra while controls at 408 nm. This spectral difference seemed to reflect specific effect of CO on cytochrome oxidase, though such trace remained for not more than a day after death.
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A method is described for the determination of delta 9-tetrahydrocannabinol (delta 9-THC) in the saliva by the use of a combination of moving-precolumn injector and glass capillary gas chromatograph with electron capture detector (GC/ECD). There were no interfering peaks due to impurities around the peak of pentafluoropropyl derivative of delta 9-THC (delta 9-THC-PFP). This GC/ECD method was linear over the range of 5-200 ng/ml of delta 9-THC-PFP. The lower detection limit was approximately 1 ng/ml. delta 9-THC content in the saliva after experimental marihuana smoking was measured by this method. It was demonstrated that for at least 4 h after smoking the level of delta 9-THC was sufficient for detection.