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Biomedical subjects

S Ishihara

Publications and source records attributed to S Ishihara.

At least 19 recordsLinked to original sources

High HTLV-I proviral DNA level associated with abnormal lymphocytes in peripheral blood from asymptomatic carriers.

The level of proviral DNA in peripheral blood mononuclear cells from a representative group of asymptomatic HTLV-I carriers in Miyazaki district, an HTLV-I endemic area in Japan, was determined by a single-cycle polymerase chain reaction method (PCR). Of 217 subjects, 26% had a high level of proviral DNA, 43% a medium level, 18% a low level, and 13% an undetectable level. In the high-DNA group, 60% had at least 0.6% abnormal lymphocytes on peripheral blood smears, significantly higher than in those with low DNA levels (19%). This association was present for men of all ages and for women under 55. Men were more than twice as likely to have abnormal lymphocytes as well as high levels of proviral DNA. These differences may reflect different host responses to the virus by sex or by the time or route of infection. This study supports the utility of PCR for molecular screening in epidemiologic studies of the natural history of HTLV-I, and may lead to the identification of those carriers who are at greatest risk of developing HTLV-I-induced malignancy.

Age Factors

Involvement of the cholinergic system in the effects of nefiracetam (DM-9384) on carbon monoxide (CO)-induced acute and delayed amnesia.

The effects of N-(2,6-dimethylphenyl)-2-(2-oxo-1-pyrrolidinyl)-acetamide (DM-9384, nefiracetam), a cyclic derivative of GABA, were investigated in the carbon monoxide (CO)-induced amnesia model in mice using the passive avoidance task. Memory deficiency occurred when mice were exposed to CO before memory was completely consolidated after training (acute amnesia), at 7 days before training and 7 days after training (delayed amnesia). DM-9384 prolonged the step-down latency in mice with CO-induced amnesia. Scopolamine blocked the anti-amnesic effect of DM-9384 on delayed amnesia that had been induced by pre- or post-training exposure to CO. Bicuculline had a tendency to antagonize the anti-amnesic effect of DM-9384, but this tendency was not significant. Under these conditions, no significant change in the activity of choline acetyltransferase and glutamic acid decarboxylase was observed in the frontal cortex, striatum and hippocampus. These results suggest that DM-9384 potentiates cholinergic neuronal function and that it may modify acquisition and/or consolidation of memory.

Amnesia

Effects of nefiracetam, DM-9384 on amnesia and decrease in choline acetyltransferase activity induced by cycloheximide.

The effects of nefiracetam, [N-(2,6-dimethyl-phenyl)-2-(2-oxo-pyrrolidinyl)acetamide, DM-9384], a cyclic derivative of GABA, were investigated in the cycloheximide (CXM)-induced amnesia animal model using the passive avoidance task. Pre-training administration of DM-9384 attenuated the CXM-induced amnesia as indicated by prolongation of step-down latency. It protected against CXM-induced inhibition of choline acetyltransferase activity in the cerebral cortex. These results suggest that DM-9384 attenuates CXM-induced amnesia by interacting with AChergic neuronal system and enhancing protein synthesis in the brain.

Amnesia

Successful graft of HTLV-I-transformed human T-cells (MT-2) in severe combined immunodeficiency mice treated with anti-asialo GM-1 antibody.

To develop an experimental model of adult T-cell leukemia/lymphoma in small animals, severe combined immunodeficiency (SCID) mice treated with anti-asialo GM-1 antibody were inoculated with MT-2 cells, a cell line transformed by the human T-cell leukemia virus (HTLV-I). Three mice injected with 4 x 10(7) cells subcutaneously or intramuscularly developed tumors at or near inoculation sites. Immunofluorescent antibody (IFA) staining for HTLV-I structural protein, p19, revealed the specific antigen in the cytoplasm of most cells from tumors and the DNA signals of HTLV-I proviral DNA were also positive in cellular DNA by polymerase chain reaction assay with HTLV-I tax gene primers, SK43/SK44. The MT-2 cells did not invade in mouse organs.

Animals

Altered glycosylation and cell surface expression of beta 1 integrin receptors during keratinocyte activation.

We studied the mechanism by which cell adhesiveness becomes activated when keratinocytes are removed from skin and placed into cell culture. Our results suggest that activation involves altered beta 1 integrin subunit glycosylation accompanied by an increase in cell surface beta 1 integrin receptors. Activated keratinocytes contained two forms of the beta 1 integrin subunit, approximately 93 kDa and approximately 113 kDa. As shown by pulse-chase experiments, the smaller represented the cytoplasmic precursor of the larger, and only the 113 kDa mature form was detected in integrin receptors expressed at the cell surface. Pre-activated keratinocytes contained beta 1 integrin subunits ranging from approximately 97 to 110 kDa. These beta 1 subunits had been processed through the Golgi, based on resistance to endoglycosidase-H treatment, and were not converted to 113 kDa subunits during subsequent cell culture. Experiments with endoglycosidase-F showed that differences in the apparent sizes of beta 1 integrin subunits observed in pre-activated and activated keratinocytes could be attributed to differences in subunit glycosylation. Smaller beta 1 subunits found in pre-activated keratinocytes, like the precursor beta 1 subunits of activated cells, appeared to be less efficient in reaching the cell surface. Overall, a approximately 10-fold increase in the level of cell surface integrin receptors occurred concomitant with the increased proportion of 113 kDa beta 1 subunits found in activated cells. Endoglycosidase-F experiments also indicated that there were changes in keratinocyte alpha subunits associated with beta 1. In related experiments, keratinocytes cultured in low Ca2+, serum-free MCDB medium for 4 days proliferated but their adhesiveness did not become activated. Therefore, keratinocyte proliferation and activation of adhesion are regulated separately. Finally, substantial activation of keratinocytes was observed when serum was added to cells cultured in MCDB with serum, indicating a role for serum factors in the activation process.

Animals

[A study of revised self-monitoring scale].

The present study attempted to construct the Japanese version of Revised Self-Monitoring Scale (Lennox & Wolfe, 1984). Factor analysis of this scale yielded two factors: 1) Sensitivity to expressive behavior of others, 2) Ability to modify self-presentation. This scale and its two factors had acceptable internal consistency: these results were almost similar with the original study. In correlational analyses with other personality measures, this scale correlated positively with both Private and Public Self-Consciousness Scale and Maudsley Personality Inventory-E Scale, but positively or negatively with some scales of Yatabe-Guilford Personality Inventory (e.g., G, S: positively. I, T: negatively.). Moreover the correlations between the two factors and the above mentioned measures provided interesting results. The availability of this scale was discussed.

Adult

[A case-control study on factors relating to discontinuation of domiciliary care for the bedridden elderly in a metropolitan area].

A case-control study was conducted to examine factors relating to discontinuation of domiciliary care for the bedridden elderly in Shinagawa-ku, Tokyo. Cases were bedridden residents aged 65 years and over who had abandoned home care and applied for admission to live in a special nursing home for the aged between April and September in 1990 after being recipients of welfare allowances for disabled elderly. Controls were bedridden residents who continued to be given home care and matched to cases by sex, age and beginning month of the receiving of allowances. Among 50 cases and 94 controls interviewed, we obtained responses from 31 cases (62%) and 60 controls (64%). The main results were as follows: 1. During the home-care period, ADL (activities of daily living) of cases, especially walking ability, deteriorated more severely than in controls. Night delirium also appeared more frequently in cases. 2. The primary caregivers of cases were older than those of controls. Remarkable differences between cases and controls were observed in the family structure, the number of family members and the number of sub-caregivers. Cases tended to live alone or live with a spouse only, and with smaller number of family members and caregivers. 3. Case lived more frequently in houses with small numbers of rooms and without rooms of their own. 4. As regards utilization of domiciliary care services, cases used dispatch of home helpers more frequently and used day services less frequently.

Aged

Analysis of factors related to hypertension in Japanese middle-aged male workers.

A total of 789 Japanese male transport service workers between the ages of 35 and 50 were used as subjects in an analysis of daily lifestyle factors related to hypertension. Multiple logistic analysis showed positive dose-response relations between hypertension and age, obesity and alcohol consumption. Age and obesity were factors having a linearly increasing odds ratios for hypertension (including borderline cases and those under treatment). Alcohol consumption of 56 g ethyl alcohol per day or more had an odds ratio about double that of those who did not drink. Smokers had 1/2 the odds ratio of non-smokers. Subjects working a 24-hr shift comprised mostly of standby duty showed a slightly lower rate of hypertension, but it was statistically insignificant.

Adult

Clinical significance of CD7-positive stem cell leukemia. A distinct subtype of mixed lineage leukemia.

Ten leukemia cases with mixed phenotype were investigated in terms of clinical characteristics and cellular origin. Three patients were infants and six patients were older children. Six of them had a high leukocyte count and a mediastinal mass was found in three cases. All but one showed hepatosplenomegaly and/or lymphoadenopathy. In spite of intensive chemotherapy, most of them responded poorly. Cytochemical analysis of their leukemic cells revealed a low percentage of positivity for myeloperoxidase reactivity (less than 25%) in two cases and electron microscopic platelet peroxidase reactivity was found in one of three analyzed cases. Phenotypically, these cells all expressed CD7, and other T-lineage-associated, B-lineage-associated, and/or myeloid-associated antigens were also detected to some extent. In addition, three cases expressed CD41 and one case expressed CD56. The T-cell receptor (TCR) genes and immunoglobulin gene were in the germline configuration in seven cases. In three rearranged cases, two showed only the TCR-delta gene rearrangement, and one had both TCR-gamma and delta gene rearrangements. Cell culture studies with 12-0-tetradecanoyl-phorbol-13-acetate (TPA) revealed differentiation to the T-lineage in two cases and to a myeloid lineage in one case. Megakaryocytic differentiation was detected in two cases in culture without TPA. These results suggest that the cells from these cases arose from stem cells capable of both lymphoid and nonlymphoid differentiation. Although the cells were heterogeneous with regard to their potency of differentiation, they have similar clinical characteristics. Because of poor prognosis, it is important to identify this type of leukemia, and allogenic or autologous bone marrow transplantation should be considered.

Acute Disease

Differential usage of delta recombining element and V delta genes during T-cell ontogeny.

We analyzed the usage of the delta recombining element (delta Rec) and six V delta genes in cell samples from 15 patients with CD3- and 10 patients with CD3+ T-cell acute lymphoblastic leukemia in an attempt to define the hierarchy of genetic events that is associated with the T-cell receptor (TCR) alpha/delta gene complex during T-cell ontogeny. Based on the deletion patterns of these genes, we surmised their relative order on chromosome 14 to be as follows: 5'-V delta 4, V delta 6, V delta 1, V delta 5, delta Rec, V delta 2, D delta 1-3, J delta 1-3, C delta, V delta 3-3'. In agreement with previous reports, V delta 1 was found to be preferentially rearranged in CD3+ samples. In CD3- samples, V delta 2 and V delta 3 rearrangements were observed at a high frequency. Incomplete V delta D delta rearrangements using V delta 2 or V delta 3, which are closest to C delta, were observed in three patients with CD3- and one patient with CD3+. These results suggest that V delta 2- and V delta 3-(Dn)D delta 3 recombinations are among the earliest recombinational events. Delta Rec was observed to be rearranged to phi J alpha on one allele. In addition, delta Rec rearrangements to J delta 1 and J alpha close to phi J alpha were also demonstrated on three alleles and one allele, respectively. Delta Rec rearrangements to J delta and J alpha other than phi J alpha also inhibit expression of the TCR delta locus. Approximately half of the alleles with J delta rearrangements showed no involvement of known V delta or delta Rec, indicating the existence of other, yet-uncharacterized V delta or delta Rec-like segments.

Antigens, Differentiation, T-Lymphocyte

Involvement of the cholinergic neuronal system and benzodiazepine receptors in alcohol-induced amnesia.

We investigated the involvement of the GABAergic and cholinergic neuronal systems and benzodiazepine (BZP) receptors in ethanol-induced amnesia using a passive avoidance task. Pretraining administration of ethanol impaired the passive avoidance response. The BZP agonist chlordiazepoxide potentiated the amnesia, while the GABA antagonists bicuculline and picrotoxin failed to affect it. The acetylcholine esterase inhibitor physostigmine partially attenuated the ethanol-induced amnesia. These results suggest that ethanol-induced amnesia is related to BZP receptors and a dysfunction of the cholinergic neuronal system.

Amnesia

Monoclonal nature of transient abnormal myelopoiesis in Down's syndrome.

Neonates with Down's syndrome occasionally show an excess of blasts in their peripheral blood. This disorder spontaneously resolves within several months and is called transient abnormal myelopoiesis (TAM) or transient myeloproliferative disorder. It has been uncertain whether the excess of blasts in TAM is a result of a clonal proliferation or a polyclonal reactive condition. The clonality of cells in females can be examined by analysis of the methylation patterns of the X chromosomes of proliferating cells using restriction fragment length polymorphism (RFLP). Using this strategy, we studied three females with Down's syndrome accompanied by TAM who showed heterozygosity in RFLP of either the hypoxanthine phosphoribosyltransferase or phosphoglycerate kinase gene. Analysis of the methylation patterns of these genes demonstrated a clonal nature for blasts in three patients. Thus, TAM is a clonal proliferative disorder. In addition, lymphocytes with a normal appearance contained in analyzed samples from these patients also showed a monoclonal pattern, suggesting that TAM may be a disorder of multipotent stem cells.

Bone Marrow

Differential effects of ursodeoxycholic acid and ursocholic acid on the formation of biliary cholesterol crystals in mice.

The preventive effect of 3 alpha, 7 beta, 12 alpha-trihydroxy-5 beta-cholanoic acid (ursocholic acid) and ursodeoxycholic acid on the formation of biliary cholesterol crystals was studied in mice. Cholesterol crystals developed with 80% incidence after feeding for five weeks a lithogenic diet containing 0.5% cholesterol and 0.25% sodium cholate. When 0.25% ursocholic acid or ursodeoxycholic acid was added to the lithogenic diet, the incidence as well as the grade (severity) of the gallstones were reduced. Plasma and liver cholesterol levels were decreased by ursodeoxycholic acid but not by ursocholic acid. Gallbladder cholesterol and phospholipid levels were decreased by both bile acids. The biliary bile acid level was decreased by ursocholic acid but not by ursodeoxycholic acid. After feeding ursocholic acid, its level in the bile was about 25% and the levels of cholic acid and beta-muricholic acid decreased. Fecal sterol excretion was not changed by ursocholic acid, but was increased by ursodeoxycholic acid. After feeding ursocholic acid, fecal excretion of deoxycholic acid, cholic acid, and ursocholic acid increased. No differences were found between mice, with or without gallstones, in plasma and liver cholesterol levels, biliary phospholipid and bile acid levels, fecal sterol and bile acid levels, and biliary and fecal bile acid composition. The results suggest that the lower incidence of crystal formation after treatment with ursocholic acid is probably by a different mechanism than with ursodeoxycholic acid. In the mouse model, ursodeoxycholic acid exerts its effect at least partially, by decreasing cholesterol absorption. Ursocholic acid is well absorbed and excreted into bile and transformed into deoxycholic acid by the intestinal microflora in mice.

Animals

Developmental process of the T-cell receptor alpha and delta gene assembly in B-cell precursor acute lymphoblastic leukaemia.

We analysed the organization of V delta genes and delta recombining element (delta Rec) in 27 children with B-cell precursor acute lymphoblastic leukaemia. Twenty-two of 54 alleles showed rearrangements of the T-cell receptor (TCR) delta locus. These rearrangements resulted either from D2D delta 3 (2 alleles) or V delta 2(Dn)D delta 3 (20 alleles) recombinations, and the other V delta and delta Rec were not rearranged. Of 23 alleles with deletion of C delta and rearrangements of J alpha, V delta 2, V delta 4 and V delta 5 appeared to rearrange to J alpha on five alleles. With regard to the relationship between the rearranged V alpha/delta and J alpha genes, gene segments 5' to V delta 2 frequently rearranged to J alpha more proximal to C alpha, whereas V delta 2 and gene segments 3' to V delta 2 showed a tendency to rearrange to J alpha distal to C alpha. Based on these findings, we suggest that the initial recombination event of the TCR-alpha/delta gene may be D2D delta 3 joining, followed by V delta 2 recombination with the D2D delta 3 complex. It was also suggested that use of V alpha/delta and J alpha/delta may depend on the distance between the involved V alpha/delta and J alpha/delta at least in B-lineage cells. These rearrangements in B-precursor cells appear to be aberrant. However, this recombinational process may be one of the normal differentiation pathways in T-lineage cells, because cells with a V delta 2(Dn)D delta 3 rearrangement were detected in 0.1-0.01% of normal peripheral mononuclear cells by the polymerase chain reaction.

Base Sequence

EGF rapidly stimulates tyrosine phosphorylation in cultured endometrial cells.

Recent increasing evidence suggests that EGF has a role in modulating the differentiated functions of human endometrial cells in an autocrine/paracrine fashion. To explore the signal transduction pathway of EGF in endometrial cells, we used cultured human endometrial cells to examine whether EGF induces tyrosine-phosphorylation. EGF phosphorylated the 175 kDa protein on tyrosine residues within 10 seconds of stimulation. EGF induced tyrosine phosphorylation at as low as 0.1 ng/ml with the maximal effect occurring at 10 ng/ml. Estradiol was shown to enhance the phosphorylation by EGF in this system. These results thus suggest that tyrosine-phosphorylation might be an important step in the signal transduction of EGF in human endometrial cells. Furthermore, the observed stimulatory action of estradiol on tyrosine-phosphorylation by EGF might provide a clue in the elucidation of the cellular mechanism of estrogen action in endometrium.

Cells, Cultured

[Experimental investigation on application of hydroxyapatite implant to alveolar ridge augmented by porous hydroxyapatite granules--histological observation on implant and implant covered with autogenous iliac bone].

UNLABELLED: The purpose of this study was to investigate the possibility of applying the hydroxyapatite implant and the hydroxyapatite implant covered with autogenous iliac bone to the augmented mandible. Three months after the extractions of the lower P3, P4, and M1 of mongrel dogs, alveolar ridge augmentations were performed widely with porous hydroxyapatite granules. After one month, the hydroxyapatite implants were inserted in the iliac bone of the same dog. Four months after the augmentation, the hydroxyapatite implant and the hydroxyapatite implant covered with autogenous iliac bone were implanted only in the augmented area with the granules of the mandible. Two months later, specimens were taken out and fixed by 10% formalin alcohol. They were embedded in polyester resin and undecalcified sections were prepared. The sections were stained with toluidine blue and observed under light microscope. RESULTS: 1) Bone ingrowth was seen in most parts of the intergranular spaces and in some spaces fibrous connective tissues were observed. 2) The hydroxyapatite implant was partly attached to the bone. But a large surface of the implant was connected to the fibrous tissues among the granules. 3) The hydroxyapatite implant covered with the iliac bone was combined with the bone of the intergranular spaces.

Alveolar Process