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The mesolimbic dopamine (DA) system has been associated with the pathogenesis of schizophrenia. Here, we show DA-containing neuronal structures of the ventral tegmental area (VTA) of an autopsy case of disorganized type of schizophrenia (75 year-old female), using tyrosine hydroxylase (TH) immunohistochemistry. A free floating method using 50-\(\mu\)m cryostat sections and three-dimensional imaging analyzer AvioVision were applied to observe wide range structures of TH-immunoreactive (-ir) neurons. TH-ir neuronal cell bodies in the VTA of the present case varied in shape and size, and TH-ir neuronal processes had irregular thickness, straightened shape, or curved shape having many corners, when compared with a control autopsy case with no detectable neurological and psychiatric diseases (64 year-old male). Mean volume of the nerve cells is diminished in the VTA of drug-naive schizophrenia. The mechanisms underlying the morphological characteristics of DA neurons of the brains with schizophrenia should further be elucidated epigenetically as well as genetically.

Lectins are proteins which specifically bind (or crosslink) carbohydrates. Recent studies have shown the importance of glycosylation in pathogenesis of diseases. We detected lectin-positive spherical deposits (SPD), 3-10 micron in diameter, in the molecular layer of dentate gyrus of hippocampus in dementia, Down' s syndrome, and schizophrenia. The present study is aimed at elucidating the nature and significances of SPD in etiology of neuropsychiatric illnesses. In schizophrenia, SPD was observed without exception, regardless of having history of pharmacotherapy. By using multi-labeling histochemical methods, single strand DNA was co-localized in hippocampal SPD of schizophrenia with lectin, including GSI-B4 for galactose, and UEA-I for fucose, suggesting that SPD formation in schizophrenia is related to apoptotic processes. The molecular basis of SPD formation should further be investigated in brains with neuropsychiatric illnesses.

Internal abdominal hernia, defined as protrusion of viscera through a defect of the mesentery, has been considered a rare clinical entity. Recent clinical reviews reported a wide range of onset age (from newborns to the elderly) and symptoms (from minimal abdominal symptoms to severe acute abdomen). Sudden and unexpected death due to internal abdominal hernia is rare in infants or toddlers, and only 4 autopsy cases had been reported previously. We report the case of a 3-month-old Japanese boy who unexpectedly died 4 h after first vomiting. Autopsy showed a wide bowel obstruction with necrosis through a congenital mesenteric defect. The larynx was filled with gastric content (milky white viscous muddy material). In the cross section of both lungs, the same material was found to be expressed from the bronchioles. We diagnosed the cause of death as asphyxiation by viscous milk/vomitus aspiration caused by bowel obstruction due to an internal abdominal hernia. In case of sudden and unexpected death of an infant, autopsy is crucial to determine the cause of death. During autopsy, it is helpful to determine the character and distribution of gastric and airway contents to confirm milk or vomitus aspiration.

Accidental death from postural or positional asphyxia can occur when an individual’s body compromises their respiration. The diagnosis of positional asphyxia is usually based on circumstantial evidence supported by the absence of other significant underlying causes of death. A female in her twenties was found dead in the so-called bridge position on a chair with wheels. Her jacket had rolled under one of the chair’s wheels. She was 159 cm in height and weighed 28.8 kg. Her body mass index was 11.4 (she was severely emaciated), and her muscles, including the rectus abdominis muscle, were thin. Her head, face, and neck were markedly congested. Her lungs, especially the upper lobes, were also congested. A small quantity of left cardiac blood was detected, which was slightly coagulated. The right cardiac blood was liquid (21 ml), and the right ventricle was slightly enlarged. It was suggested that the circulation from the lungs to the heart had been restricted. Toxicological tests detected psychoactive agents in the deceased’s blood and urine. The concentration of one of them, tofisopam, was slightly higher than normal. It was suggested that the effects of tofisopam and the deceased’s poor physical condition had impaired her motility, trapping her in an abnormal body position, ‘the reverse jack-knife position’. Therefore, her manner of death was considered to be accidental positional asphyxia. We should be aware that chairs with wheels can occasionally cause such accidents.

A female in her 30s was found dead after a fire. She was severely emaciated and had been diagnosed with anorexia nervosa (AN) about 5 years ago, but had not been treated recently. Therefore, we investigated not only her cause of death but also her condition of AN. Some of her organs weighed less than normal although no clear lesions were observed. In the pituitary gland, the number of follicle-stimulating hormone-immunopositive cells was markedly decreased although a normal number of thyroid-stimulating hormone-positive cells were detected. A histological examination of the ovary suggested that she had been suffering from amenorrhea. The thyroid gland was atrophic, and marked variations in follicle size were observed. Because we could not obtain enough volume of her blood for endocrinological examinations, we tried to investigate her endocrinological condition by immunohistochemistry. Immunohistochemical staining detected decreased triiodothyronine immunoreactivity and normal thyroxine immunoreactivity. The adrenal glands were also atrophic. Based on these findings, it was considered that she had been suffering from AN at the time of her death. The autopsy and other findings revealed that she had died of burning with carbon monoxide intoxication. J. Med. Invest. 63: 305-309, August, 2016.

Tuberous sclerosis complex (TSC) is a genetic multisystem disorder characterized by widespread hamartomas in several organs, including the brain, heart, skin, eyes, kidney, lung, and liver. Rhabdomyoma is the most common cardiac tumor diagnosed in fetuses, neonates and infants, and is closely linked to TSC. Here we describe an autopsy case of right ventricular rhabdomyoma in TSC. The deceased was a 3-month-old male infant, and TSC with a cardiac tumor had been diagnosed before his death. Since the cardiac tumor had not been physically blocking the blood flow, he had not undergone surgical intervention. At autopsy, the patient’s height was 62 cm and his body weight was 6 kg. The heart weighed 37.3 g and the right ventricle was filled with the tumor. The tumor measured 2.1 cm × 1.6 cm, being a fusion of multiple tumors with several attachment sites to the myocardium. Histologically, the tumor was diagnosed as a rhabdomyoma, and was positive for mammalian target of rapamycin (mTOR). The brain weighed 795.0 g, without hydrocephalus. The cut surface of the brain revealed multiple cortical tubers and subependymal nodules. Through screening for the TSC1 (hamartin) and TSC2 (tuberin) genes, a nonsense mutation, c.1108C>T:p.Gln370∗, was detected in the TSC2 gene. Immediate cause of death was determined to be ventricular obstruction by a cardiac rhabdomyoma with insidious growth. This case highlights the need for forensic pathologists to perform a complete autopsy to determine the cause of sudden death with cardiac tumor, including genetic examination.

With increased interest in fitness and health care, jogging has become more popular as an exercise to promote health. However, sudden cardiac death during sports or exercise has also been reported. Some apparently healthy elderly individuals take up sports for both recreation and health improvement based only on completion of a questionnaire, without undergoing medical evaluation. We report the case of a 66-year-old Japanese man who suddenly died of acute ischemic heart disease during first-time jogging. He collapsed an hour after starting. A trainer promptly started cardiopulmonary resuscitation. An automated external defibrillator (AED) was applied, and defibrillation was attempted once by bystanders. However, he remained in cardiopulmonary arrest until he reached the emergency department, where he was pronounced dead. The autopsy found concentric hypertrophy of the left ventricular wall without fibrosis or degeneration, atherosclerotic changes in the coronary arteries, and severe lung congestion. We diagnosed death from acute myocardial ischemia. We suspect that many healthy elderly individuals have provoked a heart attack by prematurely attempting moderate or vigorous exercise, as in this case. The elderly require comprehensive medical assessment before exercise can be started. Moreover, this case shows that an AED is not always helpful. J. Med. Invest. 64: 184-186, February, 2017.

BACKGROUND: It has been pointed out in Japan that criminal punishment in domestic homicide cases, especially in fatal child abuse cases, tends to be more lenient than in public homicide cases that occur outside the home. In recent news accounts of fatal child abuse cases, however, the media has reported that court-imposed sentences have tended to be stricter every year. METHODS: Using the online databases of three major Japanese newspapers, we collected articles about fatal child abuse cases that had been published from January 2008 to December 2009. We analyzed these articles to determine, whether a tendency towards tougher penalties, as was put forward by the media, actually exists at present time in the criminal system in Japan. RESULTS: We found 24 cases, out of which 20 involved only one offender and 4 involved two offenders. These 28 offenders comprised nine biological fathers, 11 biological mothers, and eight other male relatives of the child victims. We found that the sentences handed down by the court clearly tended to be more lenient for female offenders. A new system of criminal jurisprudence, the so-called saiban-in system wherein citizens serve as "lay judges" in criminal trials involving serious crimes, was implemented in Japan at the start of 2009. Each, district court has gradually adopted this new system after a preparation period of approximately five years starting in 2004. CONCLUSIONS: Many figures in the Japanese media predicted that the gap between social expectations and court sentences for sanction against domestic homicide cases would be filled with the present transitional period of the Japanese criminal system. However, the present study found no significant difference in the laws regarding sentencing in fatal child abuse cases before and after the preparation period of the saiban-in system. Language: en

Toluene is one of the most widely used organic solvents and is commonly recognized as a noxious substance inducing chronically toxic damage to neural, hepatic and renal functions in the workers engaged in printing and painting. Although hepatic cells are generally considered to be vulnerable and susceptible to various organic solvents, particularly chloroform and other halogenated hydrocarbons, the hepatotoxic effects of aromatic hydrocarbons including toluene have not yet been sufficiently characterized. In particular, it still seems unclear whether toluene itself can directly act on hepatic cells, inducing toxic damage to their metabolism and function. To assess the toxic effect of toluene inhalation on rat liver, immunohistochemical analyses of the histological markers for hepatic damage were carried out in animals exposed subacutely to toluene vapor. The immunoreactivities of heat shock proteins (HSP-70 and HSP-90) and cytochrome P4502E1 (CYP2E1) in the liver were analyzed to assess the hepatotoxic damage induced by toluene inhalation, and the expression of these histological markers was shown to be substantially enhanced by the subacute exposure to toluene vapor. Toluene inhalation was furthermore shown to enhance the immunoreactivities of alpha-smooth muscle actin (alpha-SMA), collagen, glucocorticoid receptors (GR) and leptin receptors (Ob-R) in the liver. Additional studies using human hepatoma HepG2 cells showed that toluene can directly induce toxic damage to cells. These findings suggest that toluene inhalation may primarily induce hepatic damage, which may be secondarily exacerbated by the activation of systemic processes possibly connected with glucocorticoids and leptin.

Aims: The relationship between suicide and disaster is an important problem but it's not clear. We conducted this study to determine whether a natural disaster affects suicide rates. Methods: We collected data on suicides during the 84 months before and the 60 months after the earthquake and compared the suicide rate in Kobe to that in Japan as a whole. We also examined what groups were significantly affected. Results: Compared with Japan as a whole, the suicide rates in Kobe significantly decreased in the 2 years after the earthquake. Conclusions: An influence on suicide rate after the disaster clearly appeared in middle-aged men. Language: en

Background Vertebrate-specific neuronal genes are expected to play a critical role in the diversification and evolution of higher brain functions. Among them, the glycosylphosphatidylinositol (GPI)-anchored netrin-G subfamily members in the UNC6/netrin family are unique in their differential expression patterns in many neuronal circuits, and differential binding ability to their cognate homologous post-synaptic receptors. Results To gain insight into the roles of these genes in higher brain functions, we performed comprehensive behavioral batteries using netrin-G knockout mice. We found that two netrin-G paralogs that recently diverged in evolution, netrin-G1 and netrin-G2 (gene symbols: Ntng1 and Ntng2, respectively), were responsible for complementary behavioral functions. Netrin-G2, but not netrin-G1, encoded demanding sensorimotor functions. Both paralogs were responsible for complex vertebrate-specific cognitive functions and fine-scale regulation of basic adaptive behaviors conserved between invertebrates and vertebrates, such as spatial reference and working memory, attention, impulsivity and anxiety etc. Remarkably, netrin-G1 and netrin-G2 encoded a genetic “division of labor” in behavioral regulation, selectively mediating different tasks or even different details of the same task. At the cellular level, netrin-G1 and netrin-G2 differentially regulated the sub-synaptic localization of their cognate receptors and differentiated the properties of postsynaptic scaffold proteins in complementary neural pathways. Conclusions Pre-synaptic netrin-G1 and netrin-G2 diversify the complexity of vertebrate behaviors and differentially regulate post-synaptic properties. Our findings constitute the first genetic analysis of the behavioral and synaptic diversification roles of a vertebrate GPI protein and presynaptic adhesion molecule family.

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.

The human striatum, especially its ventral part, the nucleus accumbens, contains numerous neurons immunoreactive for aromatic l-amino acid decarboxylase (AADC, the second-step monoamine synthesizing enzyme, =DDC: dopa decarboxylase), but not for tyrosine hydroxylase (TH, the first-step catecholamine synthesizing enzyme) or tryptophan hydroxylase (TPH, the first-step serotonin synthesizing enzyme) (Neurosci Lett 232 (1997) 111–114). These AADC (+)/TH (−)/TPH (−) neurons are named as D-neurons (Jaeger CB, Ruggiero DA, Albert VR, Joh TH, Reis DJ. Immunocytochemical localization of aromatic-l-amino acid decarboxylase. In: Bjorklund A, Hokfelt T, editors. Classical transmission in the CNS, Part I, Handbook of chemical neuroanatomy, vol. 2. Amsterdam: Elsevier, 1984. pp. 387–418). The nucleus accumbens is one of the brain regions that is involved in the pathogenesis of schizophrenia. We examined the distribution of striatal D-neurons using AADC immunohistochemistry and postmortem brains obtained by legal and pathological autopsies (nine controls (27–75 years old) and nine schizophrenics (32–78 years old), postmortem interval to fixation (PMI): 2–30 h). Because the number of AADC-positive neurons per section had a tendency to reduce in the case with longer PMI, we analyzed specimens of five controls (27–64 years old) and six schizophrenics (51–78 years old) in which the PMI was less than 8 h. The number of AADC-positive neurons was reduced in the striatum of schizophrenics compared to that of controls. The reduction was significant in the nucleus accumbens ( P t -test). D-Neurons might be involved in the pathogenesis of schizophrenia. Further studies using sex-, age- and PMI-matched controls are essential.

We report two cases of monostotic Paget's disease which were effectively treated with bisphosphonate. Case 1 was a 60-year-old female. Medical examination revealed high alkaline phosphatase (ALP) levels making her visit our clinic. Hematological examination showed high levels of ALP isozyme 3 and bone metabolism markers, and bone scintigraphy demonstrated strong accumulation of 99mTc on the skull. With the diagnosis of monostotic Paget's disease of the skull, treatment with bisphosphonate (etidronate) was started. The response to etidronate was good and after 12 weeks of treatment, the ALP levels decreased to about 26% of the levels before treatment, without the appearance of any symptoms or lesion development. One year and three months later, ALP increased again, and etidronate administration was resumed. However, four years after the diagnosis of the disease, etidronate became ineffective and oral administration of alendronate, a stronger bisphosphonate, was started at 5 mg/day. The patient responded favorably to the bisphosphonate and is still under observation. Case 2 was a 71-year-old female. High ALP levels were found during the follow-up of type 2 diabetes, and the case was diagnosed as monostotic Paget's disease of the pelvis based on bone metabolism markers and bone scintigraphy. Etidronate treatment at 200 mg/day resulted in the improvement of bone metabolism markers and bone scintigraphy findings. When she died of colon cancer twelve months later, with no marked progress of the Paget's disease of bone observed clinically.

Lectins are proteins which specifically bind (or crosslink) carbohydrates. Recently, importance of glycosylation has been emphasized in pathogenesis of diseases. Lectin-positive spherical deposits (SPD), 3-10 micron in diameter, have been detected in the molecular layer of dentate gyrus of hippocampus in schizophrenia, dementia, Down's syndrome, and aged individuals. In schizophrenia, SPD was observed without exception, regardless of having history of pharmacotherapy. By using multi-labeling histochemical methods, single strand DNA was co-localized in hippocampal SPD of schizophrenia with lectin, including GSI-B4 for galactose, and UEA-I for fucose, suggesting that SPD formation in schizophrenia is related to apoptotic process. The molecular basis of SPD formation should further be investigated in brains with neuropsychiatric illnesses.

Several lines of evidence suggest that a partly genetically controlled serotonergic dysfunction is involved in the biological pathogenesis of suicide. In this study, we measured tryptophan hydroxylase (TPH) immunoreactivity as a pre-synaptic marker, and serotonin receptor 2A (5HT2A receptor) density as a post-synaptic marker in the serotonergic system in 10 postmortem brains of suicide victims. We also examined whether TPH gene polymorphisms (A218C and A-6526G polymorphisms) could affect TPH immunoreactivity and 5HT2A receptor gene polymorphism (A-1438G polymorphism) could affect 5HT2A receptor density in 28 postmortem brain samples. No significant differences were found in TPH immunoreactivity or 5HT2A receptor density between suicide victims and controls. The AA genotype of the A218C polymorphism of the TPH gene showed higher TPH immunoreactivity along with lower 5HT2A receptor density than did any other genotypes in the postmortem brains of both suicide victims and controls. Our findings suggest that the A218C polymorphism of the TPH gene can be expected to provide new insights not only for neurobiological studies of suicide, but also for research into the behavioral characteristics that may be associated with serotonergic dysfunction.

Synaptic cell adhesion molecules are increasingly gaining attention for conferring specific properties to individual synapses. Netrin-G1 and netrin-G2 aretrans-synaptic adhesion molecules that distribute on distinct axons, and their presence restricts the expression of their cognate receptors, NGL1 and NGL2, respectively, to specific subdendritic segments of target neurons. However, the neural circuits and functional roles of netrin-G isoform complexes remain unclear. Here, we use netrin-G-KO and NGL-KO mice to reveal that netrin-G1/NGL1 and netrin-G2/NGL2 interactions specify excitatory synapses in independent hippocampal pathways. In the hippocampal CA1 area, netrin-G1/NGL1 and netrin-G2/NGL2 were expressed in the temporoammonic and Schaffer collateral pathways, respectively. The lack of presynaptic netrin-Gs led to the dispersion of NGLs from postsynaptic membranes. In accord, netrin-G mutant synapses displayed opposing phenotypes in long-term and short-term plasticity through discrete biochemical pathways. The plasticity phenotypes in netrin-G-KOs were phenocopied in NGL-KOs, with a corresponding loss of netrin-Gs from presynaptic membranes. Our findings show that netrin-G/NGL interactions differentially control synaptic plasticity in distinct circuits via retrograde signaling mechanisms and explain how synaptic inputs are diversified to control neuronal activity.

p15 is a novel fungal protein which induces neurite outgrowth and neuronal differentiation of PC12 cells. In the present study, we report molecular cloning, functional expression and characterization of the gene encoding p15. The deduced amino acid sequence suggested that p15 is synthesized as a precursor with 31 extra amino-terminal amino acids including a putative signal sequence, and 20 carboxy-terminal amino acids, in addition to the 118 amino acids-long mature region with neurite-inducing activity. From the poly(A)(+) RNA prepared from the producing fungal strain, a cDNA fragment encoding the mature region of p15 was amplified and His(6)-tagged recombinant p15 was produced in Escherichia coli. The recombinant protein purified by a single step on Ni(2+) agarose column chromatography exhibited comparable specific activity as native p15 in the PC12 neurite extension assay. The effect of His(6)-p15 was blocked by nicardipine, suggesting that Ca(2+) influx through the L-type Ca(2+) channels is essential for its neurite-inducing activity. In addition, mutational analysis of His(6)-p15 demonstrated that both intramolecular disulfide bonds are essential for its biological activity.

Several lines of evidence indicate that a serotonergic dysfunction is involved in the biological susceptibility to suicide. Recently, the A-1438G polymorphism of the serotonin 2A (5-HT2A) receptor gene has been suggested to be associated with suicide, but the results are inconsistent. We examined whether the A-1438G polymorphism of the 5-HT2A receptor gene was associated with suicide itself using 151 Japanese completed suicides. No significant difference in genotype distribution or allele frequencies of the polymorphism was found between the completed suicides and the comparison group. We conclude that the A-1438G polymorphism of the 5-HT2A receptor gene is not likely to have a major effect on the biological susceptibility of suicide.

A cytosolic alanyl aminopeptidase (AAP-S) was purified to homogeneity from human liver cytosol. The molecular weight of the purified enzyme was calculated to be approximately 98,000 on TOF-MS and 90,000 on SDS–PAGE in the presence of β-ME. These findings suggest that the enzyme exists as a monomeric form in human liver cytosol. The enzyme rapidly hydrolyzed the substrates Ala-, Lys- and Phe-MCAs, and moderately hydrolyzed Met-, Leu-, Tyr- and Lys–Ala-MCAs at pH ranging from 7.5 to 8.0. The order of the K cat / K m values of AAP-S at the optimal pH was Arg->Arg–Arg->Met->Leu->Lys->Phe->Lys–Ala->Tyr->Ala-MCAs. It was strongly inhibited by bestatin, leuhistin, actinonin, amastatin, 1,10-phenanthroline, DFP, PCMBS, Zn 2+ , Cd 2+ , Co 2+ , Cu 2+ , Hg 2+ and puromycin. AAP-S was approximately 80 times more sensitive than human seminal plasma AAP (aminopeptidase N, membrane type). The amino acid sequence of the first 60 residues of AAP-S was highly homologous with the N-terminal amino acid sequence of the rat liver puromycin-sensitive enkephalin-degrading aminopeptidase. These physicochemical properties and findings indicate that AAP-S from human liver cytosol is identical to those of other puromycin-sensitive aminopeptidase(s). Furthermore, with immunohistochemistry the enzyme was strongly stained in the cytoplasm of liver cells and renal tubules, and was ubiquitously localized in various human tissues.

Several lectins were used to study the localization of glycoconjugates in brain of elderly people and patients with Alzheimer type dementia (ATD) and Down's 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.

Tyrosine hydroxylase(TH) is the first step and rate−limiting catecholamine−synthesizing enzyme. Recently, TH neurons lacking higher step catecholamine−synthesizing enzyme(s) have been described in several regions of the human brain. The present study was conducted to examine whether such non−catecholaminergic TH neurons exist in the human medullary C1 adrenergic region. We quantitatively analyzed neurons immunoreactive for TH, aromatic L−amino acid decarboxylase(AADC) or phenylethanolamine−N−methyltransferase(PNMT) in the C1 region of four autopsy cases(24−64 y.o., both sexes), all without detectable neurological and psychiatric diseases. In all of the cases, the numbers of TH−immunoreactive(−ir) neurons were largest. The numbers of AADC−ir neurons were generally larger than those of PNMT−ir neurons. Double staining of TH and PNMT confirmed the existence of TH(+)/PNMT(+) neurons and TH(+)/PNMT(−) neurons. It is indicated that the human C1 adrenergic region contained non−catecholaminergic TH neurons, the significance of which remains to be elucidated.

Post-mortem brain tissue was obtained from 28 patients with brain disorders, of which 15 had clinically diagnosed schizophrenia, 6 Alzheimer type dementia, 5 dementia with tangles and 2 cases of Down's syndrome. The controls were 22 cases from autopsies without brain disorders or with no known episodes of brain disorder. The tissues were stained for the detection of carbohydrate deposits in the hippocampal formation, using lectin, immunohistochemical and conventional staining methods. The staining revealed the existence of spherical deposits in the inner and middle molecular layers of the dentate gyrus in the hippocampal formation which contained fucose, galactose, N-acetyl galactosamine, N-acetyl glucosamine, sialic acid, mannose and chondroitin sulfate. The number of the deposits was higher in patients with brain disorder such as schizophrenia, Alzheimer type dementia, dementia with tangles or Down's syndrome, and in some aged individuals, in comparison to those in younger individuals. No deposits were detected in a few younger or aged individuals. Spherical deposits 3-10 microm in diameter may be an immature form of the corpora amylacea, since they were similar in the histochemical characteristics with lectin, immunohistochemical and conventional staining methods. However, differing staining ability by hematoxylin, periodic acid Schiff's reagent and antibodies against the intracellular degraded proteins such as ubiquitin and tau-protein was observed. The antibodies against ubiquitin and tau-protein showed clear reactivity with the corpora amylacea and no reactivity with spherical deposits, indicating that the corpora amylacea has an intracellular origin and spherical deposits an extracellular matrix origin. The results obtained in this study indicate that not only neuronal degeneration but also unusual glycometabolism in neurons may disturb the neuronal function and cause brain disorders, and that spherical deposits may cause dysfunction of the neuronal network in the dentate gyrus of the hippocampus which is closely linked with recognition and memory functions.

We report a case of a seventy-year-old woman with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) and adrenal insufficiency induced by Rathke's cleft cyst. She experienced nausea, vomiting, diarrhea, and headache and disturbance of consciousness induced by hyponatremia at a serum sodium level of 100 mEq/l. In spite of severe hyponatremia, urinary sodium excretion was not suppressed and serum osmolality (270 mOsm/kg) was lower than urine osmolality (304 mOsm/kg), and arginine vasopressin (AVP) remained within normal range. SIADH was diagnosed because she was free from other diseases known to cause hyponatremia such as dehydration, cardiac dysfunction, liver dysfunction, renal dysfunction, hypothyroidism, and adrenal insufficiency. Cranial computed tomographic (CT) scan and cranial magnetic resonance (MR) imaging showed a cystic lesion of approximately 2 cm in diameter in the pituitary gland. These images suggested that the cystic lesion was a Rathke's cleft cyst, which was the cause of SIADH. Water restriction therapy normalized her serum sodium concentration and improved her symptoms. After one year, she suffered from general fatigue, appetite loss, fever, and body weight loss (5 kg/2 months). She had neither hypotension nor hypoglycemia, but her serum sodium level was low and serum cortisol, ACTH, and urine free cortisol were very low. Therefore, secondary adrenal insufficiency was suspected and diagnosed by stimulation tests. After start of hydrocortisone replacement therapy (10 mg/day), her symptoms disappeared. In conclusion, Rathke's cleft cyst should be kept in mind as a potential cause in a patient with SIADH, hypopituitarism, and/or adrenal insufficiency.

We demonstrated intense serotonin (5-HT) 2A receptor immunoreactivity in the human ventral tegmental area (VTA) using by a recently raised antibody against 5-HT2A receptor. The substantia nigra (SN) neurons also showed 5-HT2A receptor immunoreactivity. Double immunohistochemistry of 5-HT2A receptor and tyrosine hydroxylase (TH) revealed many neurons doubly labeled by 5-HT2A receptor and TH in the VTA and SN. It is suggested that activity of human midbrain dopaminergic neurons might be strongly regulated via 5-HT2A receptors at the level of their originating nuclei.

Immunoreactivity for aromatic L-amino acid decarboxylase (AADC), the second step dopamine-synthesizing enzyme, was found immunohistochemically in neurons of the human anterior cingulate cortex (ACC). Most of these neurons were located in layers V and VI and subcortical white matter; a small number were occasionally found in layer III. Double immunohistochemistry for tyrosine hydroxylase (TH: the first step dopamine-synthesizing enzyme) and AADC revealed that no neuronal cell bodies in the ACC were doubly immunostained for TH and AADC, suggesting that these TH-only- or AADC-only-immunoreactive neurons were not dopaminergic. AADC neurons in the human ACC might transform L-DOPA to dopamine, droxidopa to noradrenaline, and/or 5-hydroxytryptophan to serotonin.

The electrophoresis of proteins and enzymes is described with respect to its application and contribution to the blood group typing in the medico-legal field. Isoelectric focusing (IEF) has many advantages over conventional electrophoresis because of minimizing band diffusion, higher resolution, phenotyping under equilibrium and non-equilibrium conditions. The difference between the resolving capabilities of IEF and conventional electrophoresis is due to the pH environment within the gel. In IEF, protein separation is accomplished in a pH gradient across a gel and the net charge of a protein is variable, whereas with conventional electrophoresis the pH is essentially constant throughout the gel and net charge of a protein is also constant. A variable net charge difference along the gel plays an important role for good separation under non-equilibrium conditions. Moreover, the appropriate addition of chemical separators increases the net charge difference of proteins under non-equilibrium conditions and provides good resolution in shorter focusing times. The chemical separators also improve the resolution of isozymes and increase the protein solubility as a result of the buffering capacity. On the other hand, a shift of isozyme patterns from the same individual can be observed depending on the kind of sample. The shift is caused by the hemoglobin in phosphoglucomutase (PGM1) typing, the actin in group specific component (GC) typing and the different isoelectric points (pI) in α-1-antitrypsin (PI) typing. Therefore, the most suitable technique should be selected based on the system used and the sample conditions.

The human basal forebrain has been known to contain many neurons immunoreactive (ir) to tyrosine hydroxylase (TH; the first dopamine-synthesizing enzyme). We examined whether these neurons might contain aromatic L-amino acid decarboxylase (AADC; the second step dopamine-synthesizing enzyme) by dual labeling immunohistochemistry and confocal laser-scanning microscopy. Neurons dually-labeled for TH and AADC were found in the anterior olfactory nucleus, olfactory tubercle and the ventral margin of the rostral nucleus accumbens. The examination in the basal forebrain of the macaque monkey also gave substantially the same results. These neurons appear to constitute an independent dopaminergic cell group in the primate basal forebrain.

We examined the effect of maternal repeated cold stress (RCS) on the development of catecholamine neurons in offspring using 8-day-old offspring and tyrosine hydroxylase (TH) immunohistochemistry. RCS was loaded to pregnant rats between days 10 and 20 after fertilization. The frontal and cingulate cortices tended to contain fewer TH-immunoreactive (-ir) fibers, and the density of TH-ir varicosities with a large size (more than 7 μm in diameter) was significantly (p

Lectins are proteins which specifically bind (or crosslink) carbohydrates. Recent studies have shown the importance of glycosylation in pathogenesis of diseases. Lectin-positive spherical deposits (SPD), 3-10 micron in diameter, have been detected in the molecular layer of dentate gyrus of hippocampus in dementia, Down’s syndrome, and schizophrenia. In schizophrenia, SPD was observed without exception, regardless of having history of pharmacotherapy. Multi-labeling histochemical methods, revealed that single strand DNA was co-localized in hippocampal SPD of schizophrenia with lectin, including GSI-B4 for galactose, and UEA-I for fucose, suggesting that SPD formation in schizophrenia is related to apoptotic process. The molecular basis of SPD formation should detailly investigated in brains with neuropsychiatric illnesses, including Alzheimer’s disease and Parkinson’s disease

Adipocere formation is well known as a later postmortem change. We collected adipocere from five male victims which had been submerged under the sea or fresh water for 1 mon to 4 yr. Fresh subcutaneous fat of a male victim who died from a cerebral contusion was used as the control. The samples were homogenized, and the lipids were extracted with chloroform and methanol followed by injection into a gas chromatograph and a gas chromatograph-mass spectrometer. We detected hydroxy fatty acids (10-hydroxyoctadecanoic acid and 10-hydroxyhexadecanoic acid) as well as 10-ketooctadecanoic acid in adipocere, but not in the control. In addition, we found for the first time a cholesterol-related peak with a molecular ion of 388 in adipocere and identified it as epicoprostanol, suggesting not only oxidation but also reduction had occurred during the formation of adipocere. In addition, we showed the time-course of epicoprostanol accumulation. The relationship between the time of adipocere formation and the characteristic lipid composition is discussed.

In the human striatum, we immunohistochemically demonstrated many neurons that were immunoreactive for aromatic L-amino acid decarboxylase (AADC; the second step dopamine-synthesizing enzyme) but not for tyrosine hydroxylase (TH; the first step dopamine-synthesizing enzyme). The number of AADC-positive neurons was especially large in the ventral striatum including the nucleus accumbens. The significance of AADC-positive neurons are discussed in relation to the acting sites of L-dopa and antipsychotic drugs.

Suicide notes (SN) are potentially valuable sources of information about the psychological states of the suicidal person. It was hypothesized that there was a significant relation between suicide rate and note-leaving rate and that the incidence of note-leaving was increased during prolonged economic recession. During 21 years (1981-2001) in Kobe, of a total of 18 558 violent deaths, 5161 were due to suicide (27.8%), with 3417 male cases (66.2%) and 1754 female cases (33.8%). For each year the annual suicide rates and note-leaving rates were calculated, and this represents the percentage of committed suicides in which SN were left, among all suicide victims. In spite of the prolonged economic slump, the note-leaving rate remained almost constant (23.4-36.2%). Pearson's correlation coefficient showed no significant correlation between suicide rate and note-leaving rates (r = 0.27, P = 0.23). The finding that the incidence of note-leaving remains constant despite increasing suicide rates may suggest that the reasons for suicide do not affect note-leaving. There are cross-cultural, ethnic, and racial variations in suicidal behaviors. Although this finding may be specific in Japan, further studies of SN are needed to help clarify the suicidal states of mind.

We studied an influence of genetic polymorphisms in the cytochrome P-450IIE1 (CYP2E1) gene on ethanol elimination rate in alcoholic patients and healthy subjects. The CYP2E1 genotype was determined by polymerase chain reaction-restriction fragment length polymorphism method for 124 alcoholics and 54 healthy subjects. There was no significant difference in the gene frequency of CYP2E1 between alcoholics and healthy control subjects. Blood ethanol concentrations in the 65 alcoholics on admission ranged from 0.32 to 4.22 mg/ml. In the patients with the c1/c2 genotype, the elimination rate was significantly correlated with blood ethanol concentration. In each of the three genotypes of CYP2E1, the patients were divided into three groups based on ethanol concentrations. The average of the ethanol elimination rate in the patients with c1/c2 having blood ethanol levels of ≥2.5 mg/ml was significantly higher than the rates in the two other groups of c1/c2. When blood ethanol levels were ≥2.5 mg/ml, the elimination rate in the patients with c1/c2 was significantly higher than that in those with c1/c1. Regardless of the CYP2E1 genotype, the elimination rate in the alcoholics was higher than that in the control subjects when blood ethanol levels were

A serotonergic dysfunction in the brain has been reported to be involved in suicidal behavior independently of the presence of a specific psychiatric disorder. Serotonin 1A (5-HT1A) receptors are known to be located on serotonergic nerve terminals and to be involved in the presynaptic regulation of serotonin release. Genetic factors partly explain the risks for suicide, and a suicide completion group is thought to be more uniform than a suicide attempt group. To explore the hypothesis that the 5-HT1A receptor-induced serotonergic dysfunction is implicated genetically in suicide, we focused on the structural polymorphisms, Pro16Leu and Gly272Asp, of the 5-HT1A receptor gene, and examined the association between suicide victims who completed suicide and these two polymorphisms. In both polymorphisms, we found no significant difference in genotype distribution or allele frequencies between suicide victims and controls. These findings suggest that neither of these two polymorphisms is associated with suicide victims and it is unlikely that the 5-HT1A receptor gene is implicated in the susceptibility to suicide. © 2002 Wiley-Liss, Inc.

The mesolimbic dopamine (DA) system has been associated with the pathogenesis of schizophrenia. Here, we examined DA-containing neuronal structures of the ventral tegmental area (VTA) of an autopsy case of disorganized type of schizophrenia (75-year-old female), using tyrosine hydroxylase (TH) immunohistochemistry. A free floating method using 50-μm cryostat sections and three-dimensional imaging analyzer AxioVision were applied to observe the wide range structures of TH-immunoreactive (-ir) neurons. TH-ir neuronal cell bodies in the VTA of the present case had irregular shape and various size, and TH-ir neuronal processes had irregular thickness and straightened shape or curved shape having many corners, when compared to a control autopsy case with no detectable neurological and psychiatric diseases (64-year-old male). The mechanisms underlying the morphological characteristics of DA neurons of the brains with schizophrenia should be elucidated epigenetically as well as genetically.