Because of the sensitivity of detection of mitochondrial DNA (mtDNA) analysis, a higher success rate from old samples, severely decomposed or charred remains is achieved with mtDNA compared with nuclear DNA. Since mtDNA is maternally inherited, the sequence analysis of human mtDNA is being used widely for evidence of mother and child relationship or a sibling case. However, the mtDNA has a higher mutation rate than the nuclear DNA. Some regions of the mtDNA appear to be nucleotide substitution in all maternally linked relatives. in this study, we have carried out simultaneous typing to 11 SNPs and 2 tandem repeat sites in the mtDNA by use of the allele-specific amplification technique from 100 mother-child pairs and 60 sibling pairs. As a result, we found 9 sample pairs, out of 160 sample pairs, which appeared to be of different band intensity of each other. In a compared analysis to the 9 sample pairs, we have concluded that the difference of band intensity is attributable to point and length heteroplasmy of the mtDNA.

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  As is often the case with motorcycle-to-car accidents, the motorcycle falls over and slides before it hits a passenger car. The resulting vehicle deformation is unique. Past experimental data is unsuitable to assume the energy absorption of this unique deformation. Therefore, the motorcycle speed estimation is difficult in this type of accident and it has been long desired to acquire experimental data which would be valid for these cases. Responding to this requirement, a series of impact tests has been carried out i.e., sliding motorcycles were deliberately crashed into the side of passenger cars at various speeds. The results have shown that the deformation of the sides of the passenger cars reflect the the speed of the motorcycle. On the other hand, motorcycle deformation is less useful for the purpose of determining the speed at which impact occurred. The deformations of the test vehicles were carefully analyzed and a numerical estimation equation has also been proposed.

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  We developed a highly sensitive and rapid ABO genotyping method based on the polymerase chain reaction (PCR) using TaqMan kit, in which three single nucleotide polymorphisms (SNPs: positions no. 261, 796 and 803) were targeted and searched with original probes. This method enabled us to confirm genotypes OO, AB (10 samples each), AO, BO, AA, and BB (1 sample each) and phenotypes A and B (10 samples each). All procedures from PCR took only 2 hours and 30 minutes. The TaqMan probe PCR method required only 1.2 ng of template DNA for ABO genotyping. It also allowed us to confirm the presence of PCR inhibitors and to estimate the quantity of template DNA by using real-time PCR, which would be utilized in subsequent examinations, as with the Multiplex STR method.

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  We established a new method using latex particles that adsorbed secondary antibody to detect the antigen on single fiber. Single fiber sample fixed on slide glass with double-sided adhesion tape was blocked by 4% Block-Ace, and reacted with 50- or 100-fold diluted primary antibody. The secondary antibody that adsorbed to latex particles captured the primary antibody combined with antigen on the fiber. Positive reaction was observed clearly on the fiber sample as adhesion of latex particles, and no reaction was observed in negative cases. It was possible to detect maximally 1,000-fold diluted saliva and 10,000-fold diluted semen on single fiber by this method. This method was able to be applied to ABO blood grouping as well. Moreover, we succeeded in simultaneous detection of antigen from body fluid and ABO blood-group substances, using different color of latex particle adsorbing each of secondary antibodies. Quality of latex particles adsorbing secondary antibody had been unchanged for at least three months since latex reagent was prepared. This method was able to be applied to forensic examination such as a test of aged body fluid stain and very small amount of body fluid stain. Therefore, it was suggested that this method has wide applicability and is useful for forensic science examination.

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  Identification of plant species is one of the clues, which is an important tool in criminal investigation. However, morphological test and DNA extraction are very difficult in minute plant pieces. Thus, it is necessary to develop a technique for classification and identification based on biological and chemical components, which does not require DNA analysis. This study analyzed the lignin of the cell wall components, which are products of secondary metabolism of a plant and also examined the species peculiarity and utility in forensic science. An analysis method of lignin is Pyrolysis-Gas chromatography (Py-GC). Analysis of trace amounts of dried plant samples using Py-GC shows results in waveform data, called pyrogram. Powdered samples each of 1.4 mg were dried, in all 22 plant species including Gramineae and Leguminosae were used for Py-GC. Thirteen characteristic peaks were chosen among a pyrogram of Gramineaes and it was converted into a bar graph and compared with other plant species. As a result, same plant species showed the similar pattern, and the same plant that was taken from a different place also showed the same pattern, and reproduction characteristics were recognized. Therefore, the possibility that difference in plant species is specific was shown by the pyrogram pattern. Even if it was the sample, in which DNA cannot be extracted, as for this method, a classification of a plant species was possible. Preprocessing of a sample is simple, as it has to only be dried. In conclusion, the application of this method in criminal investigation will involve swiftness and also forms a simple classification method for plant species.

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  The mechanism of starting a fire from a kerosene fan heater by misusing of gasoline is similar to that from a wich-type kerosene heater. However, a combustion control device and many safety devices of a kerosene fan heater are different from those of a kerosene heater. Therefore, it is necessary to elucidate an outbreak of fire risk by these differences.
  In this work, an outbreak of fire risk of a kerosene fan heater by misusing of gasoline was examined through combustion experiments. Then, we confirmed that the mechanism of starting a fire from a kerosene fan heater require fuel leakage as well as a kerosene heater. The difference of the structure of a kerosene fan heater influenced the outbreak of fire mechanism. And a flame sensor that is one of the safety devices was found to detect misusing of gasoline. Moreover, the GC analysis of each component of a burnt-out kerosene fan heater was found to be effective to prove misusing of gasoline.

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  Analytical method for insulin was developed using high performance liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS). The ESI-MS conditions were optimized for determining human insulin and analogues such as lispro, aspart, glargine. Insulin was also detected for samples such as insulin needles. All insulin analogues and human type were detected as polyvalent ions, and the molecular ion peak was converted by deconvolution. Detection limits of insulins were 0.1 unit/ml (3.8 μg/ml) in scan mode, 0.002 unit/ml (77 ng/ml) in SIM mode. Aspart, glargin and human type of insulins were able to be distinguished by the established LC/ESI-MS method. However, human type was not able to be distinguished from lispro type, because molecular weight was the same. Insulin was detected from the used needle which was left outside for two months.

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  Analysis of trace amounts of crystalline impurities such as sodium chloride (NaCl) and diacetylmorphine hydrochloride (heroin) in seized methamphetamine hydrochloride (MA·HCl) powders was carried out using synchrotron radiation X-ray diffraction (SR-XRD) at beam line BL19B2 in SPring-8 for the scientific drug investigation. The analytical results were compared with those obtained by the analysis method of using conventional XRD instrument. About one milligram of the MA·HCl powder was packed in a glass capillary tube with 0.3 mm inner diameter, and exposed to the monochromatic SR X-ray beam. The diffraction lines from the seized MA·HCl powder were detected and recorded using a imaging plate in a Debye Sherrer camera.
  The peak width at half height for the SR-XRD at BL19B2 was found to be about half of that for the conventional XRD. Therefore small diffraction lines from trace impurities could be clearly separated from the other lines from major components. Some sharp lines originating from trace amounts of heroin and NaCl in a milligram quantity of the seized MA·HCl samples could be confirmed using the SR-XRD, whereas these lines could not be identified clearly by the conventional XRD. The advantages of the SR-XRD are simultaneous detection of trace crystalline impurities, including both organic and inorganic materials, in small amounts of MA·HCl samples, and rapid analysis with high angle resolution. This method should be very effective for the discrimination of illicit MA·HCl samples containing trace impurities.

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  Screening methods for blister agent hydrolysis products in an aqueous sample using capillary electrophoresis (CE) have been developed. Thiodiglycol, the hydrolysis product of mustard gas (HD) was analyzed by micellar electrokinetic chromatographic (MEKC) mode with direct ultraviolet detection at 200 nm, and the limit of detection (LOD) and within-day repeatability were 1.2 μg/ml and ≤2.8%, respectively. N-Ethyldiethanolamine, N-methyldiethanolamine and triethanolamine, which are the hydrolysis products of nitrogen mustard 1, 2 and 3 (HN-1, 2, 3) were analyzed by capillary zone electrophoretic mode using cation complexing reagent with indirect ultraviolet detection at 214 nm, and LOD and within-day repeatability were 150 ng/ml, ≤9.7%, 135 ng/ml, ≤8.2%, 90 ng/ml, ≤14.2%, respectively. It was concluded that hydrolysis products of HD and HN-1, 2, 3 in aqueous solution can be determined by the above-mentioned two kinds of CE methods, respectively.

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  We report on two forensic cases suspected in connection with anaphylactic shock. Both the patients died just after an anesthetic administration during the operation. The blood histamine and tryptase concentrations of the victims were measured by GC-MS after ethylchloroformyl derivatization and fluoroimmunoassay, respectively. In the first case, the histamine and tryptase concentrations were 58 μg/ml and 25 ng/ml, respectively, were both significantly higher than the normal blood levels. In the second case, neither concentration exceeded the normal level.

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  A urine sample obtained from a phenothiazine user was examined to identify metabolites which caused false-positive result on a screening test for methamphetamine in urine based on the Simon's color test. The urine sample was processed by liquid-liquid extraction under mild alkaline condition, and the extract was analyzed by TLC using Simon's reagent as a visualization reagent. Two blue or violet spots were appeared on the TLC plate by this reagent, and were identified as norchlorpromazine sulfoxide and norpromethazine sulfoxide by mass spectrometry after purification of the spots. Both of these two compounds possess an aliphatic secondary amine moiety, and are known as the metabolites of chlorpromazine and promethazine. These metabolites in the above urine sample were also confirmed by HPLC and LC/MS analysis. A control urine sample spiked with these metabolites gave positive result on the above screening test, indicating that these metabolites were the substances which caused false-positive on the test.

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Wrong:(Received 16 October 2006; accepted 5 February 2006)
Right:(Received 16 October 2006; accepted 5 February 2007)

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