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Biomedical subjects

S Hashimoto

Publications and source records attributed to S Hashimoto.

At least 433 records · Page 24Linked to original sources
PubMed1997

Gene transfection of mouse primordial germ cells in vitro and analysis of their survival and growth control.

We evaluated electroporation, liposome-mediated transfection, and the calcium phosphate (CaPO4) coprecipitation method for gene transfection of mouse primordial germ cells (PGCs) in culture as a prelude to the investigation of molecular mechanisms of the germ cell development. We found that electroporation severely damaged PGCs, and the efficiency of liposome-mediated transfection was very low. In contrast, using the CaPO4 coprecipitation method, 18% of PGCs transfected with plasmid pSV-LT expressed simian virus 40 large tumor antigen (SV 40 T-Ag) transiently. However, we did not detect any effects on the proliferation and survival of PGCs obtained from the embryonic gonads at 11.5 days postcoitum (d.p.c.) during 2 days of culture after the transfection. PGCs isolated from the 11.5-d.p.c. gonads change from spread- to round-shape and exhibit growth arrest during a few days of culture, and these rounded PGCs quickly disappear from the culture. We found that the transfection and expression of Bcl-XL or adenovirus type 2 E1B 19,000-molecular-weight protein (E1B 19K) significantly promoted the survival of PGCs and retarded the disappearance of rounded PGCs from the culture system. These results suggest that the Bcl-XL or E1B 19K can prevent the apoptosis of PGCs and inhibit the cell death of the rounded PGCs in culture.

Adenovirus E1B Proteins↗
PubMed1997

Enhancement of macrophage colony-stimulating factor-induced growth and differentiation of human monocytes by interleukin-10.

Interleukin-10 (IL-10) has been reported to be a negative cytokine for monocytes/macrophages. In the present study, we showed that IL-10 is rather a positive cytokine and augments the growth and differentiation of human monocytes stimulated with macrophage colony-stimulating factor (M-CSF). Highly purified adherent human monocytes were cultured for 7 days with M-CSF in the presence or absence of IL-10. The number of recovered cells increased in the culture of monocytes with M-CSF + IL-10 compared to the culture with M-CSF alone. IL-10 alone was not enough to maintain the survival and differentiation of monocytes into macrophages. Morphological change cultured in M-CSF was also accelerated by addition of IL-10, and macrophages cultured in M-CSF + IL-10 were more elongated compared to macrophages cultured with M-CSF alone. Binding of 125I-M-CSF to monocytes incubated with M-CSF + IL-10 was about 1.7-fold higher than that to monocytes incubated with M-CSF alone. In accordance with the binding study, Northern blot analysis showed that the levels of the expression of c-fms, M-CSF receptor, mRNA in macrophages cultured in M-CSF + IL-10 were higher than that in macrophages cultured in M-CSF alone. Macrophages cultured in M-CSF + IL-10 expressed higher level of Fc gamma RI, II, III, and showed augmented Fc gamma receptor mediated phagocytosis. The former also produced higher level of H2O2 and O2-, when stimulated with zymosan, and of IL-6 when stimulated with lipopolysaccharide compared to the latter. These results taken together suggest that IL-10 augments the growth and differentiation of human monocytes cultured in M-CSF.

Antigens, Surface↗
PubMed1997

Elevation of soluble CD23 in sera from patients with infectious mononucleosis.

CD23 is induced in B cells upon infection by Epstein-Barr virus (EBV) and a soluble form (soluble CD23: sCD23) is found in culture supernatants from EBV-transformed B cell lines. Based on these observations, we measured serum sCD23 levels in patients with infectious mononucleosis (IM) caused by EBV infection. Sera from patients with IM at the time of diagnosis contained more sCD23 than sera from normal control subjects. Changes in serum sCD23 levels during the course of disease showed that serum sCD23 levels were elevated at the time of diagnosis and they decreased to the normal levels during the convalescent phase defined by the improvement of symptoms of IM. These results indicate that the elevated levels of sCD23 were observed at the acute phase of IM and may be useful in diagnosing IM.

Adolescent↗
PubMed1997

Fas/Fas ligand expression and induction of apoptosis in chondrocytes.

OBJECTIVE: To examine the expression of Fas/Fas ligand and the role of this ligand/receptor interaction in the regulation of apoptosis in normal human articular chondrocytes and in osteoarthritis (OA) cartilage. METHODS: Normal and OA human knee cartilage and cells isolated from these tissues were tested for Fas expression by flow cytometry. Induction of apoptosis by antibody to Fas was analyzed by DAPI staining and electron microscopy. RESULTS: Treatment of freshly isolated normal human articular chondrocytes with an agonistic Fas antibody induced apoptosis in a subpopulation (approximately 20%) of the cells. Apoptosis induced by anti-Fas was not dependent on nitric oxide (NO), and anti-Fas also did not induce NO production. Analysis of isolated cells demonstrated similar levels of Fas expression on normal and OA chondrocytes (28% and 32%, respectively). In normal articular cartilage, Fas-positive cells were located mainly in the superficial and midzones. In contrast, in fibrillated OA cartilage, surface layers were partially absent and Fas-expressing cells were also detected in the deeper layers. Fas ligand messenger RNA was not detectable in resting or activated normal or OA chondrocytes. Analysis by electron microscopy showed the nuclear and cytoplasmic changes typical of apoptosis in cultures treated with antibody to Fas. CONCLUSION: A subpopulation of chondrocytes expresses Fas and is susceptible to Fas-induced apoptosis. Fas-mediated chondrocyte apoptosis may contribute to cartilage degradation in arthritis.

Antibodies↗
PubMed1997

Ciliary neurotrophic factor immunoreactivity in rat intramuscular nerve during reinnervation through a silicone tube after severing of the rat sciatic nerve.

The immunoreactivity of ciliary neurotrophic factor (CNTF) and S100 was studied in the degenerating and regenerating intramuscular nerves after the sciatic nerve was severed. The sciatic nerves of male Wistar rats were transected at the midpoint of the thigh, and silicone tubing was used to obtain effective reinnervation. The strong immunoreactivity of CNTF and S100 was observed in the Schwann cell cytoplasm of intramuscular nerves (IMN) and at the neuromuscular junction (NMJ) on the control sections. The CNTF immunoreactivity gradually became weak and indistinct in the Schwann cell cytoplasm after the operation. However, it was recognized again in the IMN at 4 weeks after the operation. On the other hand, the S100 immunoreactivity was continuously observed except at the NMJ through the denervating and reinnervating period. At 12 weeks after the operation, the strong immunoreactivity of both CNTF and S100 was observed again. These findings suggest that the amount of CNTF protein decreased in Schwann cells of the IMN and NMJ during the denervating period and increased during the reinnervating period in proportion to the number of remyelinated Schwann cells after severing of the sciatic nerve. They also suggest that CNTF was more highly correlated than the S100 protein with the reinnervation activity of Schwann cells.

Animals↗
PubMed1997

Neurotoxicity of serum components, comparison between CA1 and striatum.

In vasogenic brain edema, the neurotoxicity of extravasated serum components may contribute to neuronal damage. In the hippocampal CA1 sector and striatum, the neurotoxicity of serum was investigated. Rat serum was prepared as follows: Serum-1: whole serum, Serum-2: ultrafiltrated through a membrane with cut-off at molecular weight (MW) 100,000. Serum-3: through a membrane with cut-off at MW 20,000, and Serum-4: through a membrane with cut-off at MW 5,000. The infusion edema model was utilized for infusion of autologous serum into the brain. The brain tissue was histologically evaluated. The level of glutamate, total protein, and albumin was also measured in the sera used for infusion. The following results were obtained: 1) CA1 neurons were more vulnerable to all infused sera than striatal neurons, 2) there was a strong cellular response in the striatal site of infusion, but only a minimal in the CA1-sector, 3) the severity of damage of CA1 correlated with the glutamate concentrations of the infused sera, 4) further, there was a relationship between the degree of striatal neuronal loss and the amount of protein and albumin present in the infusate. It is, therefore, concluded that the neurotoxic properties of vasogenic edema fluid are also affected by specific features of the brain region of its extravasation. In addition, the pathological mechanisms associated with irreversible damage of neurons might be different in the CA1-sector and the striatum.

Animals↗
PubMed1997

Frequency of apoptosis relates inversely to invasiveness and metastatic activity in human colorectal cancer.

The frequency of apoptosis was determined in 102 cases of human colorectal cancer. The results were correlated with the frequency of cell proliferation and with clinicopathological characteristics such as degree of differentiation, invasiveness and metastasis. As a marker of apoptosis, intranuclear DNA strand breaks were localized with in situ nick translation (ISNT). As a marker of proliferation, proliferating cell nuclear antigen (PCNA) was localized immunohistochemically. The numbers of nuclei positive with ISNT and for PCNA per 1,000 nuclei on tissue sections were obtained. The labelling indices were compared with clinicopathological characteristics for each tumour. The ISNT labelling index of well differentiated colon carcinomas was higher than that of poorly differentiated carcinomas. Among similarly differentiated cancers, ISNT L.I. of colon carcinomas classified as Dukes A was higher than Dukes B/C, and L.I. of carcinomas which did not metastasize to lymph node or liver was higher than that of carcinomas which metastasized. The PCNA labelling index did not correlate with any of the clinicopathological characteristics or with the ISNT labelling index. The data suggest that apoptosis indices severe as a marker of tumour progression.

Aged↗
PubMed1997

A technique of minilaparotomy-assisted vaginal hysterectomy.

In minilaparotomy-assisted vaginal hysterectomy, the operation begins vaginally by opening the peritoneal folds and ligating the uterine vessels and uterosacral ligaments. Minilaparotomy is then performed for myomectomy, cutting the fallopian tubes and the utero-ovarian ligaments and detaching any adhesions on the anterior peritoneal angle. In 26 women who underwent this procedure, the feasibility rate was 100% and no intraoperative complications or postoperative morbidity was observed (except in one case of ovarian bleeding), indicating that vaginal hysterectomy assisted by minilaparotomy is a feasible approach for hysterectomy in the setting of large myomas, myomas with adhesions caused by endometriosis or previous pelvic surgery, and adenomyosis.

Adult↗
PubMed1997

Effects of the co-administration of 5-HT1A receptor antagonists with an SSRI in conditioned fear stress-induced freezing behavior.

The effects of the co-administration of the serotonin (5-HT) 1A receptor antagonists NAN-190 or (+)-WAY100135 with a selective 5-HT reuptake inhibitor (SSRI) citalopram on conditioned fear stress (CFS)-induced freezing behavior, which is the animal model of anxiety, were examined. The inhibitory effects of co-administration of NAN-190 (0.1-10 mg/kg) with citalopram on CFS-induced freezing were potent; in particular, at 0.1 and 0.25 mg/kg, NAN-190 significantly enhanced the effect of citalopram alone. At 0.1 mg/kg, (+)-WAY100135 also markedly enhanced the inhibitory effect of citalopram on freezing behavior. These findings suggest that 5-HT1A receptor antagonist, particularly at low doses, enhances the antifreezing effect of citalopram by blocking the autoreceptor-mediated negative feedback mechanisms of the 5-HT neuron. These experimental results concur with clinical findings that 5-HT1A receptor antagonist pindolol potentiates the effect of 5-HT reuptake inhibitors.

Animals↗
PubMed1997

Characterization and sensitivity to interleukin 2 and interferon alpha of leukemic cells from a patient with large granular lymphocytic leukemia associated with chronic active Epstein-Barr virus infection.

A patient presented with chronic large granular lymphocyte leukemia associated with chronic active Epstein-Barr virus infection (CAEBV). Cell cycle analysis revealed a minimal growth compatible with chronic lymphocytic leukemia After 5 months of treatment, the patient died from acute transformation of the leukemia. Cell harvested during chronic phase were analyzed for sensitivity to interleukin 2 (IL-2) and interferon alpha (IFN alpha) in vitro by means of surface phenotyping and cell cycle assay. IL-2 induced remarkable growth of the cells, whereas IFN alpha did not confer a growth advantage. Since IFN alpha was expected to have no growth induction effect on the leukemia cells, it was administered to the patient to treat the CAEBV.

Antineoplastic Agents↗
PubMed1997

Bufalin reduces the level of topoisomerase II in human leukemia cells and affects the cytotoxicity of anticancer drugs.

When human leukemia HL-60 cells were treated with 10(-7) M bufalin, the amounts of both topoisomerase (topo) II alpha and II beta and the activity of topo II decreased markedly and were almost undetectable 18 h after the start of treatment. The level of topo II mRNA started to decrease immediately after the start of treatment with bufalin, with a subsequent decrease in the amount of topo II alpha protein. These changes preceded the fragmentation of DNA, a typical feature of apoptosis. The results suggest that bufalin caused a marked decrease in the steady-state level of topo II alpha mRNA, which led to a decrease in the amount and activity of the enzyme and to the induction of apoptosis. A reduction in the level of topo II alpha by bufalin was also observed in other lines of human leukemia cells such as ML1 and U937. The results were exploited to potentiate the effects of cisplatin and retinoic acid (RA) on HL-60 cells: pretreatment of HL-60 cells with 10(-7) M bufalin for 6 h increased the inhibitory effects of cisplatin and RA on cell growth and enhanced the induction of cell death.

Antineoplastic Agents↗
PubMed1997

Generation of reactive oxygen species is required for bucillamine, a novel anti-rheumatic drug, to induce apoptosis in concert with copper.

Rheumatoid arthritis (RA) is considered to be a proliferative disorder of synovial tissue, which is composed of macrophage-like, fibroblast-like and dendritic cells. Bucillamine (BUC) is a novel disease-modifying anti-rheumatic drug, which is a structural analogue of cysteine. Some of the pharmacological actions of BUC have been shown to depend on the generation of reactive oxygen species (ROS) in the presence of copper. In this study, we examined whether BUC in concert with copper can induce apoptosis via generation of ROS. THP.1, a human monocytic cell line, was used as surrogate for synovial cells. We observed that BUC plus copper can induce THP.1 to undergo apoptosis, as evidenced by the presence of DNA degradation, which is preceded by ROS generation and increase in membrane permeability. Moreover, catalase rescued THP.1 from BUC-mediated cell death, indicating that generation of ROS is essential for the induction of apoptosis Red blood cells (RBC), probably acting as a scavenger of ROS, also rescued THP.1 from cell death mediated by BUC plus copper. Collectively, we suggest that ROS derived from BUC in the presence of copper may suppress the outgrowth of rheumatoid arthritis synovial cells in vivo through the induction of apoptosis.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1997

Effects of hormone replacement therapy on serum amyloid P component in postmenopausal women.

The pentraxin serum amyloid P component (SAP) is a 9.5Sz1-glycoprotein and it has recently been found to be deposited in atherosclerotic lesions or neurofibrillary tangles, which are related to the aging process and Alzheimer's disease. The level of SAP was measured by micro single radial-immunodiffusion. Sample sera were obtained from 420 healthy humans, from newborn to 86 years old. The changes in SAP during the menstrual cycle were investigated in 6 women that were 20-21 years. Fifty of the postmenopausal women, suffering from climacteric symptoms, were administered either conjugated estrogen (E), or dehydroepiandrosterone (DHEA). The SAP levels increased with age, being 1.12 +/- 0.82 mg/dl (means +/- S.D.) in neonates, and 6.15 +/- 0.92 mg/dl in persons over 80 years. The SAP level in the females between 15 and 49 years (3.32 +/- 0.95 mg/dl) was significantly (P < 0.001) lower than that in the males in the same age group (5.19 +/- 1.25 mg/dl). The SAP level in the follicular phase was significantly (P < 0.01) lower than that in menstrual phase (menstrual: 4.36 +/- 0.90 mg/dl versus follicular: 2.61 +/- 0.99 mg/dl). In the post-menopausal women that were administered E (1.25 mg/day), the SAP decreased significantly (P < 0.001) from the prelevel of 5.64 +/- 1.40 mg/dl to 4.26 +/- 0.98 mg/dl on the 14th day. In the postmenopausal women that were administered DHEA (60 mg/day), the SAP increased rapidly from the prelevel of 4.97 +/- 0.76 mg/dl to 6.17 +/- 1.20 mg/dl on the 21st day. SAP seems to be a marker that can monitor the effect of hormone replacement therapy.

Adolescent↗
PubMed1997

Prediction of growth sensitivity of acute promyelocytic leukemia cells to granulocyte colony-stimulating factor using 7AAD/PY during administration of all-trans retinoic acid.

We discussed utility of cell cycle and phenotypic analysis of acute promyelocytic leukemia (APL) cells using 7AAD/PY for the prediction of efficacy and risks of all-trans retinoic acid (ATRA) and granulocyte colony-stimulating factor (G-CSF) administration to patients with APL. Serial changes in phenotype and cell kinetics of APL cells from two patients were analyzed during ATRA administration. CD15 and CD11b were expressed on the APL cells in vivo as neutrophil maturation markers, while growth activity of the cells was decreased during ATRA administration. Using 7AAD/PY, changes in phenotype and cell kinetics were clearly detected after 2 days of cultivation with ATRA and/or G-CSF. In one patient, APL cells harvested from marrow during the first 3 weeks of ATRA administration showed distinct growth sensitivity to G-CSF ex vivo, and the cells harvested after a 4-week exposure to ATRA appeared to have lost this sensitivity. In this patient, G-CSF could be safely administered after 4 weeks of ATRA therapy. 7AAD/PY analysis is useful for predicting growth sensitivity of APL cells to G-CSF during ATRA administration.

Adult↗
PubMed1997

Familial aggregation and covariation of diseases in a Japanese rural community: comparison of stomach cancer with other diseases.

PURPOSE: We investigated familial aggregation as well as familial covariation of diseases by means of a questionnaire survey dealing with family histories of stomach cancer, stroke, hypertension, diabetes and tuberculosis as well as life style among 2,769 inhabitants of a rural community (84% of census population). METHODS: The strength of familial aggregation was shown by an odds ratio (OR) that compared the number of families in which siblings suffered from one of the above diseases among families in which at least one parent suffered from it, and among families in which neither did. Probands were divided into two groups for analysis: an under-55 "young group," and a 55-and-older "old group." RESULTS: The OR for stomach cancer was lowest and insignificant in the young group, and significant (2.2, p < 0.05) only in the old group. The OR for stroke, hypertension, and tuberculosis was 4.5-5.1 (p < 0.05) in the young group but decreased to 2.3-3.2 in the old group. Diabetes increased from 3.9 to 5.7 (p < 0.05) with advancing age. Age-related OR trends were not affected by exposure to cigarette smoke in the past. Stomach cancer showed a borderline familial covariation with diabetes and a borderline inverse covariation with hypertension. Hypertension showed a familial covariation with stroke and diabetes. CONCLUSIONS: Among the investigated diseases, familial aggregation was weakest for stomach cancer. The results suggest that stomach cancer may share a common familial etiologic factor with diabetes and hypotension.

Adult↗
PubMed1997

Objective evaluation of pain in various spinal diseases: neuropeptide immunoreactivity in the cerebrospinal fluid.

A quantitative analysis was performed of substance P-like immunoreactivity (SPLI) and of beta-endorphin-like immunoreactivity (beta-ENDLI), in the cerebrospinal fluid (CSF) in various diseases. The results reported to date have not been consistent. The purpose of this study was to investigate whether or not the concentration of SPLI or that of beta-ENDLI in CSF demonstrated any potential for assessing the degree of subjective pain in various spinal diseases. SPLI in CSF was measured by radioimmunoassay in 158 patients with a spinal disease; involving 57 patients with a lumbar disc herniation (LDH), 38 with lumbar canal stenosis (LCS), 46 with cervical myelopathy (CM) and 17 with cervical radiculopathy (CR), and also in 20 healthy controls. beta-ENDLI in CSF was measured in 25 of these same patients; involving 12 with LDH, seven with LCS and six with CM, and also five of the same controls. The concentration of serum SPLI was also measured in 50 of these 158. The severity of pain was self-evaluated by each patient using a linear visual analogue scale (VAS). Their Japanese Orthopaedic Association (JOA) score was also calculated objectively using the clinical findings. Correlations were investigated among the concentrations of SPLI and beta-ENDLI in the CSF and the VAS and JOA clinical assessments of these patients. The concentration of SPLI in CSF was significantly higher in various spinal diseases than in control (P < 0.05), and was correlated with the severity on the VAS and with the JOA score. However, beta-ENDLI was not correlated with either the VAS or the JOA score. We conclude that the measurement of the SPLI concentration in CSF has the potential for assessing objectively the severity of pain associated with various spinal diseases.

Activities of Daily Living↗
PubMed1997

TNF-alpha regulates GM-CSF-, IL-3- or M-CSF-induced Fc epsilon RII/CD23 gene expression and soluble Fc epsilon RII release by human monocytes.

The authors examined the regulatory effects of tumour necrosis factor-alpha (TNF-alpha) on granulocyte macrophage colony stimulating factor (GM-CSF)-, interleukin-3 (IL-3)- or macrophage colony stimulating factor (M-CSF)-induced gene expression of the low affinity receptor for IgE (Fc epsilon RII) on human monocytes and GM-CSF-, IL-3- or M-CSF-induced soluble Fc epsilon RII (sFc epsilon RII) release from monocytes. The expression of GM-CSF-, IL-3- or M-CSF-induced Fc epsilon RII on the surface of monocytes was reduced by TNF-alpha. The present analysis was designed to examine whether or not TNF-alpha could suppress GM-CSF-, IL-3- or M-CSF-induced Fc epsilon RII messenger RNA (mRNA) expression and enhance the release of sFc epsilon RII induced by these cytokines. The addition of TNF-alpha to monocyte cultures with GM-CSF, IL-3 or M-CSF significantly reduced Fc epsilon RII expression on the surface of monocytes and significantly increased sFc epsilon RII release from monocytes. These results suggest that TNF-alpha-dependent reduction of GM-CSF-, IL-3- or M-CSF-induced Fc epsilon RII expression on the surface of monocytes resulted, at least in part, from the suppression of Fc epsilon RII mRNA and the enhancement of sFc epsilon RII release.

Antibodies, Monoclonal↗