The purpose of this study was to evaluate the reliability of a craniometric sex estimation method proposed by Hanihara (1959) by using the latest cranial samples. A total of 80 males and 43 females with the current Japanese measurement data [NRIPS data] were collected from the archives kept in the National Research Institute of Police Science, Japan. Cranial samples used ranged in birth year from 1920 to 1979 (late Taisho to mid Showa era) and was restricted to subjects who were over 19 years of age at death.
  The means of NRIPS data were compared with those of Hanihara‘s data concerning eight measurement items (Maximum cranial length, Maximum cranial breadth, Basion-bregmatic height, Bizygomatic breadth, Upper facial height, Bigonial breadth, Mental height, Ramal height). In addition, to assess the accuracy of the sex estimation function, the NRIPS data were applied to seven discriminant functions reported by Hanihara.
  Results indicated that the means of the NRIPS were significantly greater in comparison to the Hanihara data except for maximum cranial length in both sexes, mental height in males and upper facial height in females. The error rate for sex estimation in male crania was relatively low (3.6-5.7%), but was extremely high in females (30.8-52.0%). With reference to several items of the discriminant functions, the error rates obtained were significantly different from the original error rates reported.
  The present study suggests that the application of the discriminant functions based on old population data may cause a decrease in the reliability of sex estimation in the current Japanese population.

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  The metabolism and urinary excretion of N-hydroxy-3,4-methylenedioxymethamphetamine (N-OH MDMA), a newly banned narcotic in Japan, were explored to confirm biotransformation of N-OH MDMA to 3,4-methylenedioxymethamphetamine (MDMA) and to discriminate between N-OH MDMA and MDMA intake in forensic urine analysis. The in vitro and the in vivo experiences were performed with human liver S9 and rats, respectively, and the resultant products or metabolites were determined using liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry.
  In both the in vitro and the in vivo experiences, MDMA and 3,4-methylenedioxyamphetamine (MDA) were detected, and the MDA levels exceeded the MDMA levels throughout the entire periods except for during 3 h after administration to the rats. This suggests the existence of the metabolic pathway(s) from N-OH MDMA to MDA not via MDMA. In urine samples from the administered rats parent N-OH MDMA and its demethylated metabolite N-hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA) with very low levels during short period after administration (≤6 h) were detected. The ratios of the urinary MDA/MDMA levels for N-OH MDMA-administered rats were higher than those for MDMA-administered rats. In addition, the determination of urinary diastereomers of glucuronidated 4-hydroxy-3-methoxymethamphetamine (HMMA), MDMA metabolite, revealed that the relative peak intensity of l-HMMA-glucuronide to d-HMMA-glucuronide was higher in the case of N-OH MDMA-administration than in the case of MDMA-administration.
  Detection of MDMA in both the in vitro and the in vivo experiences suggests that N-OH MDMA intake will result in MDMA excretion also in human urine. To discriminate between N-OH MDMA and MDMA intake the following view points would be applicable in urine analysis: 1) detection of N-OH MDMA and/or N-OH MDA, 2) MDA/MDMA ratio, and 3) peak intensities of diastereomeric HMMA-glucuronides.

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  Comprehensive analytical procedures for the unequivocal determination of a newly encountered drug “N-hydroxy-3,4-methylenedioxymethamphetamine (N-OH-MDMA)”, which is expected to be chemically unstable and thermally decomposed to MDMA during the analyses, have been investigated by using various qualitative analyses including color tests, thin layer chromatography (TLC), infrared spectroscopy (IR), gas chromatography/mass spectrometry (GC/MS), liquid chromatography/mass spectrometry (LC/MS), liquid chromatography/tandem mass spectrometry (LC/MS/MS) and capillary electrophoresis/mass spectrometry (CE/MS). Stability of N-OH-MDMA in aqueous solutions with several pH values and its recovery throughout the liquid-liquid extraction with ethyl acetate (EA) were also examined. Both the color tests and TLC suggested that Simon's reagent and Rf values were helpful for discrimination of N-OH-MDMA and MDMA. The IR spectra of both N-OH-MDMA hydrochloride and MDMA hydrochloride showed a sufficient difference, and the IR spectrum of N-OH-MDMA oxalate, could be identified by some of its specific peaks. GC/MS has demonstrated that both free base and its trifluoroacetyl derivative were easily decomposed to MDMA and other related products in the GC injection port, though trimethylsilyl derivatization prevented its pyrolytic disproportionation to MDMA. On the other hand, LC/MS, LC/MS/MS and CE/MS procedures were found to be reliable techniques for determination of N-OH-MDMA without its thermal and chemical decomposition. Based on the stability studies, N-OH-MDMA proved stable in acid solution, while it was expected to be transformed to isoquinoline-type compounds in neutral solution and readily decomposed to its relevant oxime in alkaline solution. Since no difference of the extraction efficiencies with EA was observed under the neutral and alkaline conditions, the extraction should be done under neutral condition to minimize its decomposition.

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  In this study, to apply immunohistochemical staining to the ABO blood grouping of non-medullated hair samples, the blocking condition and chromogenic substrate of the current protocol were modified. When 3,3',5,5'-tetramethylbenzidine was used as a substrate, the cortex of the longitudinally sectioned non-medullated scalp hair was positively stained parallel to the hair fiber; however, non-specific staining were shown in the medullae of medullated scalp hair. Non-specific staining was improved when blocking was performed using 10% normal rabbit serum at 37°C for 1 h. In the blind trial, the ABO blood group of all medullated and non-medullated scalp hair was successfully determined by immunohistochemical staining modified in this study. The biotin-conjugated ulex europaeus I suitable for this method was also validated. In conclusion, it is thought that ABO blood grouping of hair samples using this modified protocol of immunohistochemical staining is useful for screening hair samples before DNA analysis.

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  In Japan, blood fingerprints found at crime scenes are photographed and transferred to a Pap or BB sheet. The transferred fingerprints are, if necessary, enhanced by a solution of tetramethylbenzidine (TMB) or leucomalachite green (LMG). The effects of TMB or LMG on short tandem repeat (STR) typing have been studied. However, the total effects of blood fingerprints detection methods on subsequent STR typing have not been investigated enough. In this study, 3 μl of blood and 20 μl of saliva samples smeared on glass slides were transferred to a Pap or BB sheet, transferred to a BB sheet and applied with TMG or LMG solution, and only sprayed with TMG or LMG solution. DNA was extracted by an EZ-1 DNA Investigator kit or by a QIAamp DNA Investigator kit. The quantity of DNA was measured by real-time PCR assay and STR typing was performed.
  The Pap sheet-transferred portions of the samples showed no STR profiles, regardless of the DNA extraction kits. On the other hand, the BB sheet-transferred portions showed full STR profiles in all the samples. The residual portions on the glass slide that had not been transferred to Pap or BB sheets, showed full STR profiles in most of the samples. The QIAamp kit had a tendency to provide a higher quantity of DNA than the EZ-1 kit. TMB hardly or slightly reduced the quantity of DNA, but full STR profiles were still obtained. The application of LMG to the BB-sheet transferred portions drastically reduced the quantity of DNA, and affected STR typing adversely. The spraying of LMG on the samples on glass slides slightly reduced the quantity of DNA, but gave full STR profiles. It was recommended that STR typing be performed from residual portions of samples that have not been transferred to Pap or BB sheets.

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  Cannabis seeds not pretreated for prevention of germination are on sale through the Internet in recent years. These seeds are illegal and show a high probability of germination. The number of arrestees, who had cultivated illegal Cannabis seeds, has increased year after year.
  Recently, we had an opportunity to examine Cannabis seeds. We carried out experiments for the proof of the germination abilities by usual and embryo-exposure techniques. The embryo-exposure technique is a speedy and effective method for germination of Cannabis seeds, especially for the weakened seeds.
  In this study, we have reported and recorded the course of germination, how to determine the germination and the growth process of Cannabis.

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  A case of increase in normalized pulse volume (NPV) to the critical item in the field Concealed Information Test (CIT), is reported. NPV is a new index of alpha-adrenergically mediated finger vascular tone, which normally decreases in response to a critical item in CIT as the result of phasic sympathetic activation, 6 to 10 sec after the onset of a CIT question. In the present case, however, NPV increased 5 sec after the onset of the critical item. Comparing this response to the other indices of the examinee and of other cases, it has been suggested that phasic sympathetic activation was followed by sympathetic inhibition. It was also suggested that parasympathetic activation or superiority of parasympathetic activity might have occurred because heart rate decreased to the critical item. Possible psychophysiological explanations of this phenomenon have been discussed.

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