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Biomedical subjects

H Ishizu

Publications and source records attributed to H Ishizu.

At least 37 records · Page 2Linked to original sources

Tau pathology in diffuse neurofibrillary tangles with calcification (DNTC): biochemical and immunohistochemical investigation.

Diffuse neurofibrillary tangles with calcification (DNTC) represents a primary and sporadic presenile dementia that is characterized by temporal or fronto-temporal atrophy with diffuse neurofibrillary tangles (NFTs), neuropil threads and Fahr-type calcification without senile plaques. We examined the tau pathology in five autopsy cases of DNTC by immunoblotting and immunohistochemistry using phosphorylation-dependent and -independent anti-tau antibodies. The pattern of staining for different epitopes of beta-amyloid (A beta) was also investigated. NFTs were immunopositive with all the anti-tau antibodies used in this study. On the immunoblots, sarkosyl-insoluble tau appeared as three major bands of 60, 64 and 68 kDa, and as a minor band at 72 kDa. The majority of extracellular NFTs were weakly immunopositive only with the antibody recognizing the 40 carboxyl-terminal of A beta in DNTC. These results suggest that Alzheimer's disease-like tau pathology could exist independently of A beta deposits in DNTC.

Aged↗

Tau-negative astrocytic star-like inclusions and coiled bodies in dementia with Lewy bodies.

To evaluate glial lesions in cases of dementia with Lewy bodies (DLB), we studied the brains of four patients with DLB. Astrocytic star-like inclusions, which resembled tufted astrocytic fibrillary tangles in shape, were found in the cortex of two of these cases. In addition, coiled bodies were found in the white matter of the cerebrum in two cases. The astrocytic star-like inclusions were immunohistochemically negative for tau protein, ubiquitin and alpha-synuclein. The coiled bodies were immunohistochemically negative for tau protein but immunopositive for ubiquitin and alpha-synuclein. These results suggest that in DLB a primary degenerative process takes place in both glial cells and neurons.

Aged↗

Elevation of neuron-specific enolase in serum and cerebrospinal fluid of early stage Creutzfeldt-Jakob disease.

OBJECTIVE: To investigate the levels of neuron-specific enolase (NSE) in serum and cerebrospinal fluid (CSF) of patients with early stage Creutzfeldt-Jakob disease (CJD). METHODS: The levels of NSE in serum and CSF were examined in 6 cases with CJD patients. The levels of NSE in CSF were measured in 8 age matched control patients with other neurological diseases and the levels of serum NSE were also measured in another 8 age matched control patients with other neurological diseases. The groups of 8 age matched control patients consisted of 1 same patient and 7 different patients in the 2 control groups both for serum and CSF. RESULTS: The level of serum NSE in CJD (17.3 +/- 7.0 ng/ml, mean +/- SD) was significantly higher than that of controls (6.5 +/- 1.6) (P < 0.02) as was the case in CSF (79.3 +/- 53.3 ng/ml) vs (9.6 +/- 2.9) (P < 0.03). CONCLUSION: Although mean NSE levels of CJD were higher in CSF than in the serum, there still is a case with higher serum NSE level than CSF. These results suggest that the mechanism of elevation of serum NSE may not be a simple leakage from CSF, and that the measurement of serum NSE level may be useful for diagnosis of early stage CJD.

Aged↗

An autopsy case of postencephalitic parkinsonism of von Economo type: some new observations concerning neurofibrillary tangles and astrocytic tangles.

An autopsied case of postencephalitic parkinsonism of von Economo type with a 71-year duration is reported. Several cases of postencephalitic parkinsonism of von Economo type have been reported in Japan but this is the first reported case from western Japan. The patient was a Japanese man who was 74 years of age at the time of death. He developed encephalitis of unknown etiology at the age of 3 years. The first symptom was antisocial behavior, which developed at 30 years of age. At the age of 40 years, the patient showed progressive parkinsonism. Neuropathological findings disclosed marked neuronal loss with gliosis in the substantia nigra, locus ceruleus, and raphe nuclei, as well as the appearance of neurofibrillary tangles in the aforementioned areas. There were also widespread tuft-shaped astrocytes (Tu-SA) in the central nervous system, including the thalamus. Tuft-shaped astrocytes are considered to represent non-reactive astrocytes because the distributions of neurofibrillary tangles (NFT) and Tu-SA are clearly different. Therefore, the primary astrocytic lesions in postencephalitic parkinsonism of von Economo type may be more widespread. Ultrastructurally, the Tu-SA consisted of straight filaments, 15 nm in width, which formed tight bundles. Ultrastructurally, NFF in this case revealed paired helical filaments but straight filaments, 15 nm in width, which were also found in the neurons of the substantia nigra.

Aged↗

Forensic study of sex determination using PCR on teeth samples.

In this study, sex determination using polymerase chain reaction (PCR) on tooth material was evaluated from the viewpoint of forensic medicine. The sensitivity of PCR for detection of the Y chromosome-specific alphoid repeat sequence and the X chromosome-specific alphoid repeat sequence was 0.5 pg of genomic DNA. Sex could be determined by PCR of DNA extracted from the pulp of 16 freshly extracted permanent teeth and dentine including the surface of the pulp cavity of 6 freshly extracted milk teeth. Sex could be determined using the pulp in all 20 teeth (10 male and 10 female) preserved at room temperature for 22 years. For the pulp of teeth stored in sea water, the sex could be determined in all 8 teeth immersed for 1 week and in 5 of 6 teeth immersed for 4 weeks. In the remaining 1 tooth, in which sex determination based on the pulp failed, the sex could be determined correctly when DNA extracted from the tooth hard tissue was examined. For teeth stored in soil, the sex could be determined accurately in all 8 teeth buried for 1 week, 7 of 8 teeth buried for 4 weeks, and in all 6 teeth buried for 8 weeks. When teeth were heated for 30 min, sex determination from the pulp was possible in all teeth heated to 100, 150, and 200 degrees C, and even in some teeth heated to 250 degrees C. When this method was applied to actual forensic cases, the sex of a mummified body estimated to have been discovered half a year to 1 year after death could be determined readily by examination of the dental pulp. In the skeletons of 2 bodies placed under water for approximately 1 year and approximately 11 years and 7 months, pulp tissues had been dissolved and lost, but sex determination was possible using DNA extracted from hard dental tissues. These results indicate that this method is useful in forensic practices for sex determination based on teeth samples.

Adult↗

Polymorphism of the D12S391 microsatellite in a Japanese population sample.

Using the polymerase chain reaction (PCR), we studied the short tandem repeat (STR) polymorphism observed at the D12S391 locus. In 350 Japanese examined, 14 different alleles ranging from 209 bp to 261 bp were detected. Allele 18 (221 bp) showed the highest frequency at 0.30. Observed and expected values of respective genotypes satisfied the Hardy-Weinberg equilibrium (chi 2 = 24.08, P = 0.24, df = 20). In addition, 18 additional sequence structures (suballeles), were detected in this study. Within the suballeles, sequence variants, in which the initial repeat of (AGAT) was replaced with (AGGT), was found in five samples. It was found that the analysis of single-strand conformation polymorphism (SSCP) before sequence analysis was useful for distinguishing these suballeles.

Alleles↗

Occurrence of ganglioside GD3 in neoplastic astrocytes. An immunocytochemical study in humans.

GD3 immunocytochemical analysis was performed in 25 human specimens obtained by autopsy and biopsy from patients with astrocytomas, anaplastic astrocytomas, cerebellar astrocytomas and glioblastoma multiforme (GM), using the ABC method. Extraction of the ganglioside fraction from GM was used for thin-layer chromatography (TLC) analysis to confirm the specificity of anti-GD3 monoclonal antibody (DSG-1). Normal astrocytes were not immunoreactive for GD3. Neoplastic astrocytes of low- to high-grade tumours were GD3 immunoreactive. In GM, the multinucleated giant cells were also immunoreactive. All immunoreactivity present was within the cytoplasm. In TLC analysis, enzyme immunostaining of gangliosides from GM with DSG-1 showed only one positive band, which had the same TLC migration rate as GD3, indicating that GD3 of the ganglioside fraction from GM is the antigen detected by DSG-1. The presence of GD3 within the cytoplasm of neoplastic astrocytes showing invasive and proliferative properties, is of considerable interest. The implications and possible significance of the presence of GD3 in the cytoplasm in glioma cells are discussed.

Adolescent↗

Demonstration of ganglioside GD3 in human reactive astrocytes.

Astrocytes are the cells that actively participate in the process of lesion repair in the central nervous system (CNS), and reactive astrocytosis of varying degrees becomes apparent with time in any pathological condition occurring in the normally developed postnatal CNS. Ganglioside GD3 (II3a(NeuAca2-8NeuAc)-LacCer, GD3) in reactive astrocytes from autopsied patients with Creutzfeldt-Jakob disease (CJD) and old cerebral infarction was investigated immunocytochemically, using mouse IgM anti-GD3 monoclonal antibody (DSG-1). Reactive astrocytes in CJD and cerebral infarction demonstrated GD3-immunoreactivity within the cytoplasm. Normal astrocytes were negative. The present data raise the possibility that GD3 in reactive astrocytes has biological implications for the properties of the cells, such as cellular motility.

Aged↗

Depression in the early stages of Pick's disease.

To better understand the nature of the symptoms of depression in the early stages of Pick's disease, we performed a retrospective study of the medical records of eight patients who were originally treated for major depressive disorders before being clinically diagnosed with Pick's disease. Six of the eight manifested psychomotor retardation and social withdrawal, seven of the eight were agitated and five of the eight showed hyperbulia too. However, only two of the eight showed melancholia or physical symptoms such as insomnia or loss of appetite. All patients were treated with antidepressants but these were not effective in relieving the symptoms of depression. The data we gathered in this study will be useful in the future for distinguishing between Pick's disease-related depression (in the early stages of the disease) and major depression.

Affect↗

Delirium in elderly patients admitted to a psychiatric hospital: causative factors and treatment.

We studied the causes and treatments of delirium in 43 patients admitted to the geriatric-psychiatric ward of a hospital in Japan. Patients studied were divided into three groups according to the type of factor responsible for their delirium. We termed these causal factors precipitating, facilitating and predisposing factors. Twenty-one patients exhibited precipitating factors, the most common of which were overmedication and poisoning. Almost all these cases were treated with psychotropic drugs. Facilitating factors were judged responsible in nine cases. In six of these, admission to a hospital was thought to be the facilitating factor. Thirteen patients who had only predisposing factors were treated with psychotropic drugs while 27% of patients without precipitating factors were successfully treated for delirium without the use of such drugs.

Aged↗

Haptoglobin genotyping by allele-specific polymerase chain reaction amplification.

We performed haptoglobin (Hp) genotyping by polymerase chain reaction using allele-specific primer-pairs. The major six genotypes of Hp were identified using this method. Among Japanese individuals living in Ehime and Okayama Prefectures, the allele frequencies were estimated to be Hp2 = 0.723 and Hp1s = 0.277. Genotyping of Hp was possible with 0.3 ng of DNA and with 0.125 microliter of blood. It was also possible with whole blood left at room temperature for a month and also with the bloodstains left at room temperature for three years. In the heated blood samples, both alleles, Hp2 and Hp1s, were detected in those heated at 100 degrees C for 2 h. In bloodstains, Hp2 and Hp1s were detected in samples heated at 100 degrees C for 2 h and 120 degrees C for 30 min. In addition, the genotype could be detected in samples other than blood such as saliva, hair roots, tissue sections and dental pulps. The present method for Hp genotyping is expected to become a useful method in forensic analysis.

Alleles↗

Improvement of sensitivity in HLA-DRB1 typing by semi-nested PCR-RFLP.

A sensitive method of HLA-DRB1 typing was devised using a semi-nested polymerase chain reaction (PCR) followed by a restriction fragment length polymorphism (RFLP) analysis (semi-nested PCR-RFLP method). The first-round amplification (30 cycles) of the semi-nested PCR was performed using DRB generic primer pairs and the second round of PCRs (20 cycles) were performed using DRB1 group-specific primers. The products of the second round PCRs were digested with restriction endonucleases for the typing of HLA-DRB1 alleles. By this method, HLA-DRB1 typing was possible from 10 pg of genomic DNA extracted from lymphocytes and from 0.5 microliter of 1,000 times diluted blood without DNA extraction. HLA-DRB1 alleles could be typed from a 2-mm long bloodstained cotton thread prepared from 10 times diluted blood and from a 2-mm thread of whole blood bloodstains stored at room temperature for 2 years. From the mixture of blood of two individuals with different genotypes, DRB1 alleles of the minor component were detected down to 1/1,000 of the major component. This semi-nested PCR-RFLP method is useful for HLA-DRB1 typing from extremely small amounts of DNA and from mixed samples.

Genotype↗

Effects of antineoplastics, antibiotics and antidiabetics on acetaldehyde metabolism after alcohol ingestion.

The main purpose of this study was to evaluate the inhibitory effects of 5-fluorouracil antineoplastics, cephem antibiotics containing the methyltetrazolylthiol (MTT) group and antidiabetics on aldehyde dehydrogenase (ALDH) activity in vivo and in vitro. In in vivo experiments, rats were given a 100 mg/kg dose of drugs (10 mg/kg for glibenclamide) orally or intraperitoneally. When each drug was administered singly immediately after an oral administration of 1.5 g/kg ethanol, only carmofur, an antineoplastic, produced marked increases in blood acetaldehyde concentrations. This action was also noted when ethanol was ingested 15 h after administration. The remaining drugs did not increase blood acetaldehyde concentrations. When rats were treated with carmofur at 12 h intervals for 3 consecutive days and were given 1.5 g/kg ethanol after the final treatment, blood acetaldehyde concentrations were elevated more significantly than with a single administration of carmofur. Furthermore, daily administration of cephem antibiotics containing the MTT group, latamoxef, cefamandole, cefoperazone and cefbuperazone, significantly increased blood acetaldehyde concentrations. Daily administration of sulfonylurea antidiabetics, chlorpropamide and acetohexamide, slightly increased blood acetaldehyde concentrations. Drugs causing increases in blood acetaldehyde concentrations when administration was combined with ethanol ingestion also inhibited ALDH activity in vitro. The results of the in vitro experiments roughly correlated with those of the in vivo experiments. The inhibitory effects of drugs on ALDH activity were in the following order: carmofur >> cephem antibiotics containing the MTT group > sulfonylurea antidiabetics.

Acetaldehyde↗

Estimating the time between drinking and death from tissue distribution patterns of ethanol.

To establish a method for estimating the time between the last consumption of alcohol and death, we examined the ethanol levels in body fluids and tissues of rats that had been orally administered 1 g/kg ethanol. We observed the following relationships between ethanol levels in the cardiac blood (blood in the heart itself), vitreous humor, and urine: cardiac blood > vitreous humor > urine at 10 min (early absorption stage); vitreous humor > cardiac blood > urine from 20 to 50 min (late absorption stage); vitreous humor > urine > cardiac blood from 60 to 120 min (distribution stage); and urine > vitreous humor > cardiac blood at 180 min (excretion stage). It was also observed that, in cases of death immediately following drinking, ethanol levels in the stomach contents are very high, and the following ratios of ethanol levels were observed: skeletal muscle to cardiac blood--less than 1; liver to cardiac blood--around 1. buccal mucosa to cardiac blood-greater than 1. These ratios at equilibrium after drinking were around 1, lower than 1 and around 1, respectively. We also measured alcohol levels in the cardiac blood, urine, vitreous humor and stomach contents of nine cadavers who had consumed alcohol prior to death. The relationships between the time since last consumption of alcohol and relative ethanol levels in these specimens were in good accordance with the results of the animal experiments.

Administration, Oral↗

Expression of amphiphysin I, an autoantigen of paraneoplastic neurological syndromes, in breast cancer.

Amphiphysin I is a 128 kD protein highly concentrated in nerve terminals, where it has a putative role in endocytosis. It is a dominant autoantigen in patients with stiff-man syndrome associated with breast cancer, as well as in other paraneoplastic autoimmune neurological disorders. To elucidate the connection between amphiphysin I autoimmunity and cancer, we investigated its expression in breast cancer tissue. We report that amphiphysin I was expressed as two isoforms of 128 and 108 kD in the breast cancer of a patient with anti-amphiphysin I antibodies and paraneoplastic sensory neuronopathy. Amphiphysin I was also detectable at variable levels in several other human breast cancer tissues and cell lines and at low levels in normal mammary tissue and a variety of other non-neuronal tissues. The predominant amphiphysin I isoform expressed outside the brain in humans is the 108 kD isoform which represents an alternatively spliced variant of neuronal amphiphysin I missing a 42 amino acid insert. Our study suggests a link between amphiphysin I expression in cancer and amphiphysin I autoimmunity. The enhanced expression of amphiphysin I in some forms of cancer supports the hypothesis that amphiphysin family members may play a role in the biology of cancer cells.

Alternative Splicing↗

A comparative review of the immunobiology of murine neuroblastoma and human neuroblastoma.

BACKGROUND: The prognosis for children with neuroblastoma (NB) remains dismal, in part because of extent of disease at diagnosis as well as resistance of tumors to conventional therapies. However, human NB exhibits many favorable traits, including the capability to mature into a more benign form or to regress spontaneously. A murine model of disease that could permit eventual genetic manipulation, so that such beneficial traits could be identified or even augmented, would be most useful. METHODS: This report details an analysis of the contemporary study of the tumor growth, metastases, immunogenicity, and immunotherapy of murine NB and compares it with known patterns of clinical NB. RESULTS: Striking similarities exist in the local tumor growth and metastatic behaviors of murine and human NB, behaviors that in part may be related to a host-tumor immunologic response. The naturally low expression of class I antigen in NB, the ability to augment that expression with cytokines, and the phenotype of the cellular and humoral immune response to NB are strikingly similar in human and murine hosts. Comparable immunotherapeutic potential exists. There have been rare and sporadic observations of spontaneous regression of an existing murine NB, unlike the more predictable regression of Evans Stage IV-S clinical NB. CONCLUSIONS: The many similar biologic, physiologic, and immunologic characteristics of human and murine NB make the murine model a valuable adjunctive aid in furthering our understanding of human disease.

Animals↗