Identification of furosemide in hair in a post‐mortem case by UHPLC‐MS/MS with guidance on interpretation

Testing for drugs in hair raises several difficulties. Among them is the interpretation of the final concentration(s). In a post‐mortem case, analyses revealed the presence of furosemide (12 ng/mL) in femoral blood, although it was not part of the victim's treatment. The prosecutor requested our laboratory to undertake an additional analysis in hair to obtain information about the use of furosemide. A specific method was therefore developed and validated to identify and quantify furosemide in hair by UHPLC‐MS/MS. After decontamination of 30 mg of hair, incubation in acidic condition, extraction with ethyl acetate, the samples were analyzed by UHPLC‐MS/MS. Furosemide was found in the victim's hair at 225 pg/mg. However, it was not possible to interpret this concentration due to the absence of data in the literature. Therefore, the authors performed a controlled study in two parts. In order to establish the basis of interpretation, several volunteers were tested (four after a single 20 mg administration and twenty‐four under daily treatment). The first part indicated that a single dose is not detectable in hair using our method. The second part demonstrated concentrations ranging from 5 to 1110 pg/mg with no correlation between dosage and hair concentrations. The decedent's hair result was interpreted as repeated exposures. In the case of furosemide analysis, hair can provide information about its presence but cannot give information about dosage or frequency of use.     

Education Reform in Japan: A Course for Lifelong Learning

Japan's current education system has its origins in postwar reform, overemphasizing individualism and underemphasizing on Japan's history, traditions, and culture, resulting in the continuing decline in scholastic, physical, and socializing ability to date. This essay reviews the IIPS proposal on educational reform, which was supervised by former Prime Minister Yasuhiro Nakasone, with the addition of the author's personal views. The IIPS proposal set the ultimate goal of education in Japan to be raising healthy people who have self-confidence and pride as Japanese who can thrive in the era of globalization. Then, the proposal presents what a Japanese should learn and how he should serve at each life stage beginning with early childhood education through the compulsory education period, adolescence and young adulthood, maturity, and into the elderly period. Moreover, the organizational reform on administrating education policy is presented, with a focus on abandoning the current board of education system.     

Statistical comparison of dog and cat guard hairs using numerical morphology

Numerical features obtained from the guard hairs of dogs and cats (total 300 hairs per dog or cat) were statistically compared, in an attempt to discriminate between them. Using hairs from each of five mongrel dogs and cats, eight measurements (length (Len), maximum width (MaxWid), cross sectional maximum diameter, cross sectional minimum diameter, cuticular thickness of the cross section and three scale counts per 100 microm length (observed at three positions: distal third (disSC), middle (midSC) and the proximal third (proSC) portions) and five indexes (hair width index (HWI), medulla index (MI), hair index, cuticle index and the difference in scale counts between the distal and proximal parts (defSC)) were examined. The range for each numerical feature overlapped each other extensively, and none of the features permitted a discrimination between dog and cat hairs, based on the values obtained. However, 12 numerical features, except for the midSC, showed a statistically significant difference between dog and cat hairs, as evidenced by a t-test. For the purpose of comprehensively comparing numerical features and statistically discriminating between dog and cat hairs, a discriminant analysis between the two were carried out using a multiple regression analysis. Four types of discriminant functions produced by combining over five numerical features were examined. Dog and cat hairs could clearly be discriminated using any of the discriminant functions. Species discrimination using the discriminant function permitted the species of a dog or cat to be determined, based on the overall morphologies of various numerical features. When experimentally collected test samples were investigated using the discriminant function using Combination-2, consisting of eight numerical features (Len, MaxWid, MI, HWI, disSC, midSC, proSC and defSC), all 10 cat hairs were correctly determined to be cat hair and 22 of 23 dog hairs were correctly identified. This discriminant function produced good results for species discrimination between dog and cat hairs.     

Regional distribution of methamphetamine in autopsied brain of chronic human methamphetamine users

We measured levels of methamphetamine and those of its metabolite amphetamine in 15 autopsied brain regions of 14 human methamphetamine users. Only slight regional differences were observed in drug concentrations among the brain areas. Although, some redistribution of the drugs probably occurred postmortem, these data suggest that methamphetamine might not be preferentially retained in dopamine-rich brain areas but is heterogenously distributed in brain of chronic human users of the drug. The possible pharmacological actions of methamphetamine in both dopamine-rich and poor brain areas of chronic drug users need to be considered.     

Forensic Potential of MMPs and CC Chemokines for Wound Age Determination

In this study, we investigated time-dependent expression of matrix metalloproteases (MMP)-2, MMP-9, chemokine CC motif ligand (CCL)-2, CCL-3, CCL-5, IL-1β, IL-6, and TNF-α mRNA at the skin injury site and sought their forensic potentials during the skin wound repair process. The tested wound ages in 42 mouse skin wounds were distributed at 0d, 1d, 3d, 5d, 7d, 10d, and 14d, respectively and then followed by real-time polymerase chain reaction. Ultimately, MMP-2 played an important role in the inflammation phase. On the contrary, MMP-9 became involved at a later phase during wound healing. Meanwhile, CCL-2 and CCL-3 were active throughout almost all of the process. However, CCL-5 mRNA had no significance. Collectively, an MMP-9/MMP-2 ratio of over 0.84 indicated that skin wound healing age was strongly 5 days or less. So elevated gene expressions of cytokines and chemokines in different phases of wound ages implied that combined exploration could make wound age determination more accurate and objective.     

Population Aging in Postwar Japan: Processes and Prospects

This paper critically reviews postwar population development in Japan and aims to identify unsolved issues, which are possibly shared by other countries, and extract lessons for the future. First, I examine long-term development on fertility and life expectancy in Japan with a brief introduction of pre-war population history. I stress that some critical issues have often been ignored in policy debates and some important new challenges are now emerging, factors that are indispensable to extract lessons for the future. Second, I turn to the official population projections and related issues which again require scrutiny. Third, I emphasize that serious scientific investigation from multiple disciplines is essential to cope with the challenges of demographic changes and that Japan should take the leading role in research for the next generations all over the world.     

Computer-assisted skull identification system using video superimposition

This system consists of two main units, namely a video superimposition system and a computer-assisted skull identification system. The video superimposition system is comprised of the following five parts: a skull-positioning box having a monochrome CCD camera, a photo-stand having a color CCD camera, a video image mixing device, a TV monitor and a videotape recorder. The computer-assisted skull identification system is composed of a host computer including our original application software, a film recorder and a color printer. After the determination of the orientation and size of the skull to those of the facial photograph using the video superimposition system, the skull and facial photograph images are digitized and stored within the computer, and then both digitized images are superimposed on the monitor. For the assessment of anatomical consistency between the digitized skull and face, the distance between the landmarks and the thickness of soft tissue of the anthropometrical points are semi-automatically measured on the monitor. The wipe images facilitates the comparison of positional relationships between the digitized skull and face. The software includes the polynomial functions and Fourier harmonic analysis for evaluating the match of the outline such as the forehead and mandibular line in both the digitized images.     

Determination of Both Fetus’ and Mother's Blood Type from an Autopsy Case Immersed in Formalin for Over 50 Years

A female fetus which had been immersed in formalin for more than 50 years was found in Japan. Because no liquid blood could be obtained, we tried to use immunohistochemistry (IHC) methods to tissue samples obtained at autopsy to identify both the fetal and mother's blood type. We detected B antigens on endothelial cells in paraffin sections of the fetal organs. Furthermore, we observed both anti‐A‐ and anti‐B‐positive red blood cells in the intervillous space, which is indicative of the mother's blood type. To our knowledge, this is the first case report on determining the blood type of both the fetus and the mother from tissue immersed in formalin for such a long time. The results suggest that IHC is valuable for the determination of ABO blood type in circumstances of long postmortem duration and unfavorable storage conditions.