In 2006, brownish-pink clandestine tablets with an “S” stamp mark were seized in several prefectures in Japan. These were reportedly sold as “Ecstasy (MDMA tablet)”, but routine analysis indicated no traces of any controlled substances or common recreational drugs. However, additional high resolution mass spectrometry and nuclear magnetic resonance spectroscopy confirmed the unusual component N-(2,4,6-trimethylphenyl)phthalimide (TMPPI). Because this compound is considered to be a derivative of Thalidomide, potential sedative and anticarcinogenic effects were expected from its structure.
  To investigate its metabolism and toxicity, administration experiments were conducted on rats. As a result, N-(o-carboxybenzoyl)-2,4,6-trimethylaniline was identified as a specific metabolite in the rats' urine. Furthermore, no symptoms were noticed with a single dose of 350 mg/kg TMPPI. Thus, TMPPI was found to have a low acute toxicity.

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  Qualitative analyses of trace elements in the glass for car lamps were performed by wavelength dispersive X-ray fluorescence analysis (WD-XRF) and synchrotron radiation X-ray fluorescence analysis (SR-XRF) for the discrimination of the car lamps. Glass samples were crushed into powder and a disc of the same shape prepared from the powder of each glass was used for WD-XRF and SR-XRF measurements under the same geometrical condition in order to make the comparison of the spectra more accurate. Comparison of peak intensities for elements (Fe, As, Sr, Zr, Sb, Ba and Pb) detected by WD-XRF was effective for the discrimination of the glasses. In SR-XRF, difference in the peak intensities of Fe, As, Sr, Zr and Pb was found in some of glass samples produced by a same manufacturer. Difference in the peak intensities for elements in glass samples was observed also in the elemental constitutions of glass samples for the same use. The differences in elemental constitution were observed also between the outer glass (bulb glass) and the inner glass (stem glass) within the same lamp. Both WD-XRF and SR-XRF are very powerful analytical techniques to discriminate glass of the car lamps. The constituent of elements obtained by these methods can provide information to some extent to assist in determining the manufacturer and specifications of the glass.

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  Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was applied to the determination of trace elements in glass fragments by ablating a rough surface of it for the forensic discrimination of extremely small glass fragments of irregular shape by comparison of elemental compositions. Two fragments were taken from each of a glass standard reference material (NIST SRM 612) and five different sheet glass samples and used to make fragments with a flat or rough surface for each of them. Mass spectra of the 5 sheet glass indicated that the comparison of concentrations for 10 elements (Co, Rb, Sr, Zr, Mo, Ba, La, Ce, Nd and Pb) would be useful for the discrimination of the sheet glass sample. The concentrations for these elements in the fragments made from the sheet glasses were determined by LA-ICP-MS using NIST SRM 612 as one-point external standard and compared with those by solution nebulization ICP-MS (SN-ICP-MS), which were validated by analyzing NIST SRM 612. Measured values by LA-ICP-MS using either a flat or rough surface showed satisfactory good agreement with those by SN-ICP-MS with no significant difference between flat and rough surfaces, which demonstrates that analytical results obtained by LA-ICP-MS was as accurate and precise as those by SN-ICP-MS. LA-ICP-MS using either a rough or flat surface allowed complete discrimination of five sheet glasses. From these results, it is strongly recommended that LA-ICP-MS should be applied to extremely tiny fragments of irregular shaped glass in actual cases without any pretreatment such as polishing, to reduce the risk of sample loss.

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  Some .38 caliber revolvers smuggled into Japan are homemade guns called Paltik which were made in Cebu Island, the Philippines. Bullets fired from a Paltik have the following general characteristics;
(1) Rifling mark is rough and unclear because of the inferior processing of the finished rifling bore.
(2) Bullet is elongated because the bore diameter is usually smaller than an authentic gun.
  But some Paltik bullets nowadays have a neat rifling mark. The diameter in the base of the lead bullet may have been changed thus having the rifling mark deformation become indistinct. In such cases, it becomes more difficult to distinguish fired bullets between those fired from a Paltik and an authentic gun.
  In a former report, the maximun rate of contraction in the base of lead bullets and reduction of the angle of twist on the rifling mark fired from an authentic gun was indicated. But because the shape of bullets fired from a Paltik whose bore diameter is smaller, the deformation characteristic becomes long and narrow, so it is necessary to discriminate the characteristics of deformation on bullets fired from a Paltik and from an authentic gun.
  In this study, .38 Special lead bullets were shot into thick steel plate in a right angle from a Paltik whose bore diameter is 8.45 mm (diameter of fired bullet is 8.58 mm). The following results were obtained and it was concluded that there were few differences in characteristics of deformed bullets between a Paltik and an authentic gun.
A) The diameter in the base of bullets was contracted up to 0.59 mm (8.58 mm → 7.99 m, 6.9% smaller) in a Paltik. On the other hand, it was 0.6 0mm (9.00 mm → 8.40 m, 6.7% smaller) in an authentic gun.
B) The angle of twist of rifling mark reduced 0.75° (6.90° → 6.15°, 11% smaller) in a Paltik. On the other hand, it was 0.79° (3.30° → 2.51°, 24% smaller) in an authentic gun.

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  Serial arsonists (N=125) were differentiated into four groups on the basis of their social independence and the degree of criminality, by using categorical principal components analysis (CATPCA). Results indicated the following. (1) Arsonists with high social independence and high criminality were aged 40 years or more in age, did not live with their parents, and tended to have criminal records for theft. (2) Arsonists with high social independence and low criminality were most frequently employed, high school graduates, married, and had no criminal record. Moreover, there were more female and mental patients in this group compared with other groups. (3) Arsonists with low social independence and high criminality were all male, compulsory education level (including high school dropouts), unmarried, not living with parent(s), and had no criminal record of theft. These arsonists tended to have relatively no criminal record of arson compared with other groups. (4) Arsonists with low social independence and low criminality were the youngest among four groups and were aged between 10 and 30 years, unmarried, lived with their parent(s), and had no criminal record. Results of log-linear analysis indicated that arsonists with high social independence tended to use a car or walk to the crime scene, whereas those with low social independence were inclined to use a bicycle. Moreover, arsonists with high criminality records tended to prepare the medium for arson in advance and drink alcohol before the offence, whereas arsonists with low criminality had a tendency to set fire to the same place repeatedly.

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  In 2004, the Food Safety Commission under the Cabinet Office of Japan decided to implement an evaluation of the risk of food poisoning by all species of bacteria that threatens the food supply. However, this statute was designed to protect food products from accidental and uncontrollable adulteration and not from the possibility of intentionally adulterated food products.
  In this report, we evaluated the viability and proliferation of highly pathogenic bacteria, Bacillus anthracis spore form, Bacillus anthracis vegetative form, Brucella melitensis, Francisella tularensis, Yersinia pestis, and Burkholderia pseudomallei, when mixed into various food products. Among the beverages tested, the viability of all pathogenic bacteria, including the spores of B. anthracis, was retained in milk; but a carbonated drink containing vitamin C inhibited the viability of B. anthracis spores by 16%. In electrolyte replenishing beverages (ion supply drinks), B. anthracis spores retained 70% viability in contrast to the vegetative form, which retained only 1.7% viability. Furthermore, when the pathogenic bacteria were cultured on the surface of various processed food products, the risk of B. pseudomallei proliferation was demonstrated as well as the risk of B. anthracis. Also, when various pathogenic bacteria were introduced into ready-to-eat packaged curry and rice, the spores of B. anthracis remained viable under all conditions over a period of 30 days, indicating a high potential risk from this organism.
  Thus, we demonstrated the risk of serious harm posed by various forms of food tampering that could be committed by persons with malicious intentions or by terrorists.

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  Rapid, efficient, and easy-to-use procedures for isolating high-quality genomic DNA or RNA of pathogenic bacteria or viruses prior to gene amplification procedures are essential for the molecular identification of biowarfare agents. In this study, we developed a rapid and simple nucleic acid extraction method for bacteria and its spore. The method is based on cell disruption using zirconia/silica beads followed by precipitation of nucleic acids with isopropanol by using a personal microcentrifuge. In spectrometric and agarose gel electrophoretic analysis, we confirmed that bacterial genomic DNA was efficiently purified from gram-negative, gram-positive bacteria and spores. We evaluated this extraction method for the application for real-time PCR-based biological detection system, R.A.P.I.D.^TM. The detection sensitivities of the R.A.P.I.D.^TM system to the samples prepared from Bacillus anthracis vegetative cells and spores by our extraction method were 100-fold higher than those by using the IT1-2-3^TM extraction kit, which is recommended by the instruction of R.A.P.I.D.^TM. In the loop-mediated isothermal amplification (LAMP) detection, which we previously developed, B. anthracis DNA in the extracts of 2×10² cfu/mL of spore suspensions could be detected. In addition, B. anthracis in 0.5% (w/v) white powder (e.g. sugar) suspension was also detected by R.A.P.I.D.^TM and LAMP. The DNA extraction method we developed here is highly efficient and applicable for nucleic acid amplification techniques. In addition, this method was able to be performed with simple devices. Therefore our method was considered to be suitable for the on-site use for detection of microbiological agents.

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  In many countries, including Japan, cultivation of Cannabis sativa (C. sativa) for drug use has been illegal and prohibited. Recently, seeds for cultivation purposes are easily available from internet shops, and then we have often been requested to identify C. sativa. The identification has conventionally been performed by morphological and chemical tests. But, it can be difficult to identify tiny and fragmenting samples as C. sativa even if these tests are performed.
  In this study, we aimed to establish a method based on DNA analysis. As an initial step, we attempted a method reported by Linacre et al, however, cross-amplification between C. sativa and Humulus lupulus (H. lupulus) with C. sativa specific primers (G and H) was observed. To avoid this cross-amplification, we designed a new primer specific for C. sativa (cp-Can) on trnL intron of chloroplast DNA. DNA samples from nine plants including C. sativa and H. lupulus were amplified using the green plant universal primer pair and the cp-Can. After subsequent agarose gel electrophoresis, C. sativa DNA showed two bands, whereas the other plant DNA showed one band, indicating the clear distinction from the other plants tested. In addition, a BLAST search with the cp-Can sequences showed no cross-activity with other plants. The present method is very simple, rapid, sensitive, and useful for the identification of C. sativa.

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  Grasses are common plant species encountered in botanical evidence because of their wide range of habitats. Presently forensic plant samples are discriminated based on morphological observation, however, some textures occasionally show similar characteristics among different species. In this study, discrimination of grass samples was carried out by comparison of rbcL partial region sequence, which is used as markers for phylogenetic and taxonomic studies. Sequence analyses were performed on 5 commercially available grass hay samples and plant fragments collected from 3 different cowsheds. The rbcL sequences of commercially available grass hay samples were characteristic to each other. All grass samples collected from the same cowshed had identical sequences, and the sequence was characteristic to each cowshed. Furthermore, different parts of the plant body could be analyzed and showed the identical sequences within the same sample. The results suggest that discrimination among grass hay samples was possible by comparison of rbcL sequences and DNA sequences used as phylogenetic and taxonomic markers will be useful for forensic discrimination of plant evidence.

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  During the identification of single fiber evidence, color evaluation is one of the most important examinations. Colors are usually first evaluated by ultraviolet-visible-microspectrophotometry (UV-vis-MSP). If possible, thin-layer chromatography (TLC) or high performance liquid chromatography (HPLC) are conducted after the extraction of dyestuffs from the single fiber. In this report, three woolen single fibers were dyed by similarly colored dyes with structurally different yellow acid dyes and were used to evaluate the discriminatory abilities of UV-vis-MSP and TLC.

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