Akio Tanegashima

Liquefied petroleum gas (LPG) poisoning: report of two cases and review of the literature.

Two autopsy cases of men who died while connecting a liquefied petroleum gas (LPG) pipe are reported. Their blood concentrations of propane (the main content of LPG) were 0.12 and 3.40 mg/100 g, respectively. The cause of death after exposure of LPG has generally been considered to be asphyxia from hypoxia. The large differences in the blood propane levels found here and reported in the literature, however, suggest that direct toxic effects of propane poisoning may be the cause of death in some cases. Propane concentrations and the cause of death are reviewed and discussed.

Estimation of stress in child neglect from thymic involution

It is difficult to evaluate the extent of stress in cases of suspected child abuse/neglect in a medico-legal autopsy. We have previously reported that stress due to abuse/neglect was found to have led to thymic involution. To elucidate the influence upon thymocytes differentiation, we compared the proportion of the thymocyte subpopulation in the thymus of a neglected child with one in an age-matched control obtained from cardiac surgery. We found that the relative number of CD4+ CD8+ double positive (DP) thymocytes decreased in the neglected child. It was presumed that the selective decrease in the number of the immature DP thymocytes with CD3- to low bcl-2low caused the thymic involution in the neglected child. It was suggested that an alteration in the proportion of thymocytes subpopulation might be used as an index of stress in cases of child abuse/neglect.

Changes in choice of method and lethality between last attempted and completed suicides: how did suicide attempters carry out their desire?

Some researchers have emphasized that, from the perspective of suicide prevention, research into the methods of suicide seemed to be particularly promising, as it has been shown repeatedly that restricting access to the prevailing method of suicide in a country will decrease suicide rates and that the lethality of the method used significantly correlated with the degree of intention to die. In this study, we examined changes in choice of method and the lethality score between the last attempted suicide (LAS) and completed suicide (CS) in 416 victims (male: 197, female: 219) to point out the tendency on their choice of method in LAS and CS. There was a significant difference in choice of suicide method between LAS and CS, and injury to themselves (33.7%) was the most common method of LAS, while hanging (37.5%) was the most common method of CS. The mean lethality score of CS method was significantly higher than that of LAS method in both sex groups, suggesting that at least one of the causes that drives suicide attempters to commit suicide finally may be the difference in the lethalities of LAS and CS. At the time of CS, suicidal victims tend to choose the same method as that of LAS again. These findings suggest that although suicide attempters tend to choose the same method, they will use a more lethal method if they change the suicide method. Interestingly, moreover, there was no sex difference in the percentage of the mean lethality score at CS.

Determination of ABO genotypes with DNA extracted from formalin-fixed, paraffin-embedded tissues.

SummaryThe gene encoding the specific glycosyltransferases which catalyze the conversion of the H antigen to A or B antigens shows a slight but distinct variation in its allelic nucleotide sequence and can be divided into 6 genotypes when digested with specific restriction enzymes. We extracted DNA from formalin-fixed, paraffin-embedded tissues using SDS/proteinase K treatment followed by phenol/chloroform extraction. The sequence of nucleotides for the A, B and O genes was amplified by the polymerase chain reaction (PCR). DNA fragments of 128 by and 200 by could be amplified in the second round of PCR, using an aliquot of the first round PCR product as template. Degraded DNA from paraffin blocks stored for up to 10.7 years could be successfully typed. The ABO genotype was deduced from the digestion patterns with an appropriate combination of restriction enzymes and was compatible with the phenotype obtained from the blood sample.ZusammenfassungDas Gen wird für die spezifischen Glykosyltransferasen kodiert, welche die Konversion des H-Antigens in das A- oder B-Antigen katalysieren, weist eine leichte, aber definierte Variation seiner allelischen Nukleotid-Sequenz auf. Auf diese Weise lassen sich 6 Genotypen unterscheiden, nach Spaltung mit spezifischen Restriktionsenzymen. Wir haben DNA von formalinfixierten, paraffineingebetteten Geweben extrahiert, indem wir SDS-Proteinase-K-Behandlung mit nachfolgender Phenol-Chloroform-Extraktion anwandten. Die Sequenz der Nukleotide für die A-, B- und O-Gene wurde amplifiziert mit Hilfe der Polymerase-Kettenreaktion (PCR). DNA-Fragmente mit 128 Bp und 200 Bp konnten in der zweiten PCR-Phase amplifiziert werden, indem ein Aliquot der ersten PCR-Phase eingesetzt wurde. Degradierte DNA von Paraffin-Blöcken, welche bis zu 10,7 Jahren gelagert waren, konnte erfolgreich typisiert werden. Der ABO-Genotyp wurde dann abgeleitet von den Mustern nach Spaltung mit einer geeigneten Kombination von Restriktionsenzyneu und war kompatibel mit dem Phänotyp, der von der Blutprobe erhalten wurde.

ABH-related antigens in human male genital tract

SummaryThe localization of ABH related antigens in human male reproductive tract was examined using monoclonal antibodies and an avidin biotin complex method. No positive reaction with blood group antibodies on spermatozoa was observed in testis and ductus epididymidis apart from erythrocytes and endothelial cells. The expression of ABH and ABH related antigens in ductuli efferentes testis, ductus epididymidis, seminal vesicle and prostate was complexly coded by a combination of H, Se, Le and X genes. The results obtained in this study indicate that the ABH antigens detected on spermatozoa of seminal stains are coating antigens and not inherent to the cell membrane, and the ABO, H, Se, Le and X genes are subjected to a tissue-dependent differential expression.ZusammenfassungDie Lokalisation (and Verteilung) der Antigene des ABH-Komplexes im Bereich der inneren Geschlechtsorgane des Mannes wurde mittels monoklonaler Antikörper unter Benutzung einer Avidin-Biotin Technik untersucht. Dabei konnten positive Reaktionen im Hoden und im Ductus epididymidis lediglich an Erythrozyten und Endothelzellen beobachtet werden. Die Expression von ABH-Antigenen in den Ductuli efferentes testis, im Ductus epididymidis, in den Samenbläschen und der Prostata wird offensichtlich komplex durch H-, Se-, Le- und X-Gene kodiert. Die Resultate der vorliegenden Untersuchungen zeigen, daß die ABH-Antigene der Spermienoberfläche offensichtlich aus der Samenflüssigkeit stammen und die ABO-, H-, Se-, Le-und X-Gene gewebsabhängig unterschiedlich exprimiert werden.

Utilization of lectin-histochemistry in forensic neuropathology: lectin staining provides useful information for postmortem diagnosis in forensic neuropathology.

We have investigated the deposition of glycoconjugates in human brain tissue with or without brain disorders. In this review we describe the application of lectin-histochemistry techniques to forensic neuropathology. Lectin staining is able to reveal several kinds of carbohydrate-related depositions in addition to the conventional degenerative changes including senile plaques, neurofibrillary tangles and corpora amylacea. The senile plaques and neurofibrillary tangles were clearly stained by Con A, PSA and GSI lectins, the corpora amylacea which is relevant to repeated brain hypoxia and mitochondrial damage was also easily detected by these and many other kinds of lectins. Amorphous spaces were detected around blood vessels and independently from blood vessels by lectin staining in the white matter from patients with brain disorders or severe edema. The white matter lesions were not considered relevant for forensic pathology, until a large group of cerebral white matter lesions were detected in the elderly with increasing frequency by modern neuro-imaging methods. The spherical deposits were newly detected by lectin staining in the molecular layer of the dentate gyrus of the hippocampal formation chiefly from patients with schizophrenia or cognitive dysfunctions.

Histochemical characteristic of perivascular space in the brain with an advanced edema

Amorphorous and colorless spaces, Virchow-Robin spaces (VRS), were often found by HE stain around blood vessels in the edematous brain. Histochemical characteristic of the enlarged VRS caused by an advanced edema and detected by lectin stain using Griffonia simplicifolia I agglutinin in the brain stem, the occipital lobe and/or the cerebellum was examined by means of immunohistochemical method. After pretreatment with formic acid or proteinase K, formalin fixed-paraffin embedded tissue sections were incubated with antibodies (ABs) against plasma proteins such as amyloid P component, Ig G, albumin (Al), apolipoprotein E (Apo E), and lactotransferrin (Lf), and cellular proteins such as ubiquitin (Ubt), Tau-protein (Tau), glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), CD68 (KP-1) and heparansulfate proteoglycan (HSG). The tissue sections were also incubated with antibodies against alanyl aminopeptidase-S (AAP-S) and alanyl aminopeptidase-N (AAP-N) without pretreatment. The VRS showed intensive reactivity with ABs against Amy P, AAP-S and AAP-N, moderate with ABs against Apo E and HSG, weak with ABs against Ig G, Al and Lf, feeble with ABs against Ubt, Tau and CD 68, and no with ABs against GFAP and MBP, respectively. Although the substances detected in VRS might be of blood plasma origin resulting from abnormalities in the blood-brain barrier, the mechanisms whereby the serum proteins and/or other substances are enriched in VRS remain incompletely understood.

Lectin-histochemical detection of degenerative glycoconjugate deposits in human brain

Several lectins were used to study the localization of glycoconjugates in brain of elderly people and patients with Alzheimer type dementia (ATD) and Downs syndrome (DS). Five kinds of degenerated or deposited materials stained clearly by lectins specific to GalNAC, Gal, Fuc, and/or Man were recognized much in ATD and DS, less in elderly peoples, in addition to the binding of the lectins to neurons. (i) Round shape deposits called corpora amylacea (CA) which consisted of various sizes of round material, existed mainly on the surface of cerebral cortex and some in white matter of the brain. They were colored by Alcian blue (AB), Aldehyde fucsin (AF) and periodic acid shiff (PAS) and weakly by Hematoxylin (H), but not by Eosin (B). They showed clear reactivity with lectins specific to GalNAC, Gal, Fuc and Gal-GalNAC. (ii) Amorphous and variform amyloid deposits existed around blood vessels in the white matter were stained by thioflavin and lectins specific to GalNAC, Gal and Fuc, but not with Man specific lectins and PAS, AB, AF and HE. (iii) Another kind of amyloid deposits which showed a similar characteristic to the previous one and were recognized mainly in white matter and independent blood vessels. These deposits were stained by thioflavin but not by PAS, AB, AF and HE and showed good reactivity with lectins specific to GalNAC, Gal, Fuc, Gal-GalNAC, Gal-GIcNAc and Man. The reactivity with lectins specific to Gal, Fuc, and Man was seen in senile plaques (iv) and neurofibrillary tangles (v). Although at present we are unable to explain the origin of these deposits, it is clear from this study that the glycoconjugates form an integral part of the degeneration in the brain. The lectin staining with GS-I is useful in the forensic pathology to diagnose brain disorders at postmortem examination, since these lectin were able to detect five types of degeneration changes and/or deposits.

Fatal acute alcohol intoxication in an ALDH2 heterozygote: a case report

On an evening in November, a 25-year-old man was found dead in his bedroom. There were many empty snap-out sheets for flunitrazepam tablets in the trash at his bedside. He had been beaten by a gang of young people earlier in the morning of the same day. At the medico-legal autopsy, although there were many bruises and/or abrasions on the whole body, only slight subdural hemorrhage was observed, and none of them was thought to be the cause of death. Flunitrazepam and its metabolites were not detected in his body fluid by gas chromatography-mass spectrometry (GC-MS). Marked lung edema and a severe congestion of organs were observed. His blood alcohol concentration from the femoral vein was 2.00 mg/ml. Fatal cases of acute alcohol intoxication usually have shown higher alcohol concentration (2.25-6.23 mg/ml). Although the genotype of aldehyde dehydrogenase 2 (ALDH2) has not previously been mentioned as a contributing factor in determining the cause of death, in this case the genotype of ALDH2 was ALDH2*1/2 and thus is important. Those who possess the ALDH2*2 gene show high concentrations of acetaldehyde (AcH) at even comparatively lower alcohol levels. Consequently, the cause of death was considered to be acute alcohol intoxication including AcH poisoning.

Ethnic differences in the expression of blood group antigens in the salivary gland secretory cells from German and Japanese non-secretor individuals.

Type 1 ABO blood group antigens (peripheral core structure: Galβ1-3GlcNAcβ1-R) are expressed mainly in endodermally-derived tissues, but are not synthesized in mesodermally-derived tissues. In the former tissues, H type 1 antigen is generated largely by α-2-l-fucosyltransferase encoded by secretor (Se) gene and acting on the terminal galactose of the type 1 precursor chain. This theory has been generally accepted, and it seems that the expression of ABO blood group antigens is absent, or expressed at a low level, in these tissues from non-secretor individuals. In this immunohistochemical study on the secretory cells of salivary glands, we found ethnic difference between German and Japanese non-secretor individuals in the expression of blood group antigens: i.e. the expression of the type 1 blood group antigens is present in these cells from Japanese non-secretor individuals but absent from German. A possible explanation is that another α-2-l-fucosyltransferase, independent of the secretor gene, is present in Japanese non-secretor individuals.