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Biomedical subjects

M Fukushima

Publications and source records attributed to M Fukushima.

At least 271 records · Page 15Linked to original sources
PubMed1995

A female heterozygous patient with Fabry's disease with renal accumulation of trihexosylceramide detected with a monoclonal antibody.

We describe a female patient with heterozygous Fabry's disease. The patient had persistent proteinuria and microhematuria but lacked any other diagnostic signs such as corneal and cutaneous involvement. Kidney tissue obtained at biopsy showed the segmentally distributed enlarged glomerular epithelia. These cells were filled with vacuolated foamy cytoplasm, which had lamellar and myelinoid structures under electronmicroscopic observation. Accumulation of trihexosylceramide (CTH) in these foamy epithelial cells was confirmed with immunohistochemical staining with the use of anti-CTH monoclonal antibody. Alpha-galactosidase activity of leukocytes was 67 nmol/mg protein/hr, which was approximately half that of the normal population (mean +/- SD, 147 +/- 65 nmol/mg protein/hr, n = 20). All of these findings were compatible with the diagnosis of heterozygous Fabry's disease. We recommend that kidney tissue biopsy specimens suggesting Fabry's disease be immunostained with anti-CTH antibody.

Antibodies, Monoclonal↗
PubMed1995

Vasoactive intestinal peptide gene expression in the rat pheochromocytoma cell line PC12.

Vasoactive intestinal peptide (VIP) gene expression was analyzed in PC12 cells. VIP mRNA was detected in PC12 cells treated with VIP or forskolin whereas no VIP mRNA was detected in the untreated cells. The induction of the VIP mRNA was enhanced by the simultaneous treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). PC12 cells stimulated with forskolin plus TPA released immunoreactive VIP. Sephadex G-50 column chromatography revealed that the immunoreactive VIP secreted from PC12 cells is comprised of multiple forms, one of which was indistinguishable from the authentic VIP. PC12 cells supported an efficient transcription from the human VIP gene promoter in a cell-specific as well as cAMP-dependent manner. These results definitely demonstrated the expression of the VIP gene in PC12 cells. VIP biosynthesis may be positively regulated by VIP in an autocrine fashion in PC12 cells.

Animals↗
PubMed1995

Effect of cytotoxic prostaglandin, delta 12-prostaglandin J2 on E-cadherin expression in transformed epidermal cells in culture.

The cyclopentenone prostaglandins (PGs), such as delta 12-PGJ2 and PGA1, are potent inhibitors of growth in a variety of cultured cells, including human epidermal cells. To clarify the mechanism of the cytotoxicity of these PGs, we examined the effects of delta 12-PGJ2 on the function and expression of E-cadherin, which plays a major role in the maintenance of intercellular adhesion, in transformed human epidermal cells in culture (HSC-1). A 12-h incubation with 5 micrograms/ml of delta 12-PGJ2 did not affect the cell-binding activity of E-cadherin expressed in HSC-1 cells. Immunoblot analysis using a monoclonal antibody specific to human E-cadherin revealed that a 12-h incubation with 5 micrograms/ml of delta 12-PGJ2 induced E-cadherin expression in HSC-1 cells. Immunofluorescence using a monoclonal antibody against human E-cadherin demonstrated that E-cadherin was localized to the cell-cell contact regions in HSC-1 cells. Following a 12-h incubation with 5 micrograms/ml of delta 12-PGJ2, E-cadherin was also detected in a uniform pattern along cell junctions, although cell morphology was changed by the presence of cytotoxic PGs. These results suggest that the cytotoxicity of cyclopentenone PGs is related, at least in part, to E-cadherin expression in transformed human epidermal cells.

Antibodies, Monoclonal↗
PubMed1995

The effect of a probiotic on faecal and liver lipid classes in rats.

The effect of a probiotic composed of Bacillus, Lactobacillus, Streptococcus, Saccharomyces and Candida species (each at 10(7-8) colony-forming units (cfu)/g rice bran), given at a level of 150 g/kg diet for 6 weeks, on lipid metabolism was examined in the faeces, serum and liver of male rats. Liver weight decreased 35% in the rats fed on a high-fat, high-cholesterol diet containing the probiotic. Total cholesterol concentration in the serum was significantly lower in the probiotic group than in the control group throughout the experimental period in rats fed on the high-fat, high-cholesterol diet, and HDL-cholesterol concentration was significantly higher (P < 0.05) in the probiotic group than in the control group which was fed for the 6 week experimental period on a basal diet. The serum VLDL+IDL+LDL cholesterol concentration in the probiotic groups were reduced compared with those of the corresponding control groups. The probiotic groups fed on the high-fat, high-cholesterol diet and the basal diet had lower hepatic cholesterol concentrations than did the corresponding control groups (P < 0.05). Hydroxymethylglutaryl coenzyme A reductase (NADPH) (EC 1.1.1.34) activity in the liver was lower in rats fed on the high-fat, high cholesterol diet with the probiotic. The neutral and acidic steroid concentrations in faeces were higher in the probiotic group than in the control group fed on the high-fat, high-cholesterol diet. Escherichia coli decreased and Bifidobacterium and Eubacterium increased in the faecal microflora of rats fed on the dietary probiotic. Lactobacillus in the probiotic groups was higher than that in the control groups. The present study shows that the probiotic promotes Bifidobacterium and Eubacterium in the faecal microflora, and reduces cholesterol levels in the serum and liver of rats.

Animals↗
PubMed1995

Effects of the lipid-saccharide complex and unsaponifiable matter from sunflowers on liver lipid metabolism and intestinal flora in rats.

The effects of the flower lipid-saccharide complex and unsaponifiable matter (1 g/kg of diet) from the sunflower on liver lipid metabolism and intestinal flora was studied in rats given cholesterol-enriched diets. After six weeks of feeding, the microsomal cholesterol concentration in the liver had been significantly reduced with the sunflower diet. The ratio of cholesterol/phospholipid was also reduced by the sunflower diet. The 3-hydroxy-3-methylglutaryl coenzyme A reductase activity of the sunflower groups was significantly lower than that of the control group. There was no significant difference in the cholesterol 7 alpha-hydroxylase activity, although this tended to increase with dietary sunflower consumption. The number of Bacillus in the cecum flora was significantly higher in the lipid-saccharide complex group than in the other groups, while Bifidobacterium and Eubacterium in the cecum flora was significantly higher in the unsaponifiable matter group when compared to the control group. These results suggest that the lipid-saccharide complex and unsaponifiable matter in the sunflower are related to liver cholesterol synthesis and intestinal flora.

Animal Nutritional Physiological Phenomena↗
PubMed1995

[A case of rubella encephalitis: rubella virus genome was detected in the cerebrospinal fluid by polymerase chain reaction].

We studied a 2-year-old boy who had been diagnosed as having rubella encephalitis by detection of rubella virus genome in his cerebrospinal fluid with the reverse transcription-polymerase chain reaction (PCR). PCR was considered to be very useful not only to make an early diagnosis of rubella encephalitis but also to be applied generally to that of various viral encephalitis.

Cerebrospinal Fluid↗
PubMed1995

[Aneurysmal bone cyst of the left 5th rib].

A case of aneurysmal bone cyst of the left 5th rib is reported. A 57-year-old male complained of palpable lump and pain on his back. Chest X ray and CT showed expansive tumor arid pathological fracture of the left 5th rib. The tumor and the adjacent segments of the 4th and 5th ribs were resected en bloc. The pathological examination showed multiple blood filled spaces and fibrous trabeculae, confirming the diagnosis of aneurysmal bone cyst. He has been followed up for 3 years and 6 months with no signs of recurrence.

Bone Cysts, Aneurysmal↗
PubMed1994

Characterization of high-affinity binding sites for diisopropylfluorophosphate (DFP) from chicken spinal cord membranes.

The delayed neurotoxic organophosphate [3H]diisopropylfluorophosphate ([3H]DFP) binds with high affinity to membrane-bound proteins from the chicken spinal cord. The DFP binding proteins were solubilized from membrane preparations, using a detergent (CHAPS). The protein(s) sites that labeled with a low concentration of [3H]DFP, e.g. 10(-10)-10(-9) M, were defined as the high-affinity binding sites. The density (or concentration) of the high-affinity binding sites in protein(s) was determined by the difference between total and non-specific binding. The high-affinity binding sites were saturable, and the maximal amount of binding sites was estimated at 400 fmol/mg protein. [3H]DFP binding to solubilized proteins was not completely reversible. Concentration-dependent curves suggested that the [3H]DFP binding sites differ from the active sites of acetylcholinesterase, butyrylcholinesterase, and neuropathy target esterase, as well as from muscarinic acetylcholine receptors. The amount of DFP binding sites after a neurotoxic dose of tri-o-cresyl phosphate (TOCP) decreased markedly in membrane preparations from the chicken spinal cord. These results indicate that a TOCP metabolite(s) interacts with the DFP binding sites in vivo. Gel filtration chromatography of the solubilized membranes indicated at least two major high-affinity DFP binding proteins with apparent molecular weights of 300 and 110 kDa. The DFP binding sites corresponding to the 110 kDa protein were insensitive to eserine, a potent anti-cholinesterase agent.

Animals↗
PubMed1994

Regulation of recombinant human granulocyte colony-stimulation factor production using herpes simplex virus 1 thymidine kinase gene.

For chronic neutropenic patients requiring long-term injection of recombinant human granulocyte colony-stimulating factor (rhG-CSF), a cellular transplantation system that can produce this cytokine stably and deliver it in a regulatory manner would be advantageous. In this study we aimed at developing a regulation system at cellular level using suicide vectors. We introduced the herpes simplex virus type 1 thymidine kinase (HSV-TK) gene into the rhG-CSF-producing NIH3T3 cells and examined if ganciclovir (GCV) treatment of the cells could control the rhG-CSF production in vitro. The cells transfected with the HSV-TK gene showed a > 100-fold increase in sensitivity to GCV compared with the parent cells, and the median inhibitory dose of GCV to the transfected cells was less than 1.6 microM. The total amount of rhG-CSF production by these cells was strongly suppressed by GCV treatment. This regulatory method may be applicable to cytokine supplement gene therapy.

Ganciclovir↗
PubMed1994

A new vector for the high level expression of chimeric antibodies in myeloma cells.

We previously reported the expression of a mouse/human chimeric anti-ganglioside GD3 antibody, KM871 (IgG1,kappa) in mouse myeloma SP2/0 cells under the control of the ecotropic Moloney virus long terminal repeat by the co-transfection of chimeric heavy (H) and light (L) chain vectors (Shitara et al. (1993) Cancer Immunol. Immunother.). To establish an efficient and high level expression system for the chimeric antibody, we did comparative study on vector systems and host cells. An improved expression vector, named 'a tandem vector, pChi641HLGM4' was constructed, in which both of the chimeric H and L chain gene transcription units and a dihydrofolate reductase (dhfr) gene transcription unit were inserted. When two kinds of mouse myeloma cell lines, SP2/0 and P3U1, were used as host cells, frequency of the incidence of antibody-producing transfectants was markedly increased by the use of the tandem vector compared with the use of the mixture of each chimeric H vector and L chain vector. To select out appropriate host cells, transfection frequency and antibody production level were compared among SP2/0, P3U1 and rat myeloma YB2/0 cells by transfection of the tandem vector. YB2/0 cell was shown to have the highest potential in both the transfection frequency and the antibody production. Introduction of the tandem vector into YB2/0 cells and the subsequent amplification with 50-200 nM methotrexate gave rise to several clones that stably secreted 70-100 micrograms/10(6) cells per 24 h of the chimeric antibody. This productivity of the antibody is one of the highest levels which have been achieved by other investigators using transfected myeloma cells. Using this system it took only 2-3 months to establish the transfectant clones which stably produced the chimeric antibody.

Animals↗
PubMed1994

Observer disagreement in histological classification of ovarian tumors in Japan.

Histological slides of 789 cases of ovarian tumor registered by the Japanese Gynecologic Oncology Group, sponsored by the Ministry of Health and Welfare, Japan, were reviewed by three certified pathologists in order to evaluate the observer disagreement among the pathologists and the reproducibility of the histological classification. In 53.0% of the cases, all observers agreed upon the diagnosis. At least two of them agreed in 88.4% of the cases. The observer disagreement was highest for low potential malignancy (borderline malignancy) tumors (44.4%), including serous, mucinous, and endometrioid tumors. The results of the study clearly indicate the necessity of establishing a slide review system in order to obtain reliable data for comparative research. Moreover, the histological criteria, especially of tumors of borderline malignancy, should be more clearly defined to obtain high reproducibility of ovarian tumor diagnosis.

Carcinoma↗
PubMed1994

Preclinical studies of antitumor prostaglandins by using human ovarian cancer cells.

Pleiotropic actions of antitumor prostaglandins (PGs) on tumor cells are reviewed including our preclinical results focused on human ovarian cancer. Regarding inhibition of cell proliferation, antitumor PGs exerts its action as a G1 blocking agent. The cyclopentenone PGs inhibit myc oncogene expression while inhibiting the cell cycle progression and results in apoptotic cell death and growth inhibition. Cyclopentenone PGs inhibit growth of various tumors transplanted to mice or nude mice and show adjuvant effects to cis-diamminedichloroplatinum(II) (CDDP). In order to elucidate a role of antitumor PGs in immune systems, relevance of effects on tumor growth with those on the immune systems are also discussed with our results.

Animals↗
PubMed1994

Sequence variation of human papillomavirus type 16 E7 in preinvasive and invasive cervical neoplasias.

Variation in the nucleotide sequence of the HPV 16 E7 gene in preinvasive cervical intraepitherial neoplasia (CIN) and invasive cervical carcinoma specimens was analyzed. Direct DNA sequencing of PCR-amplified products with primers different from those used for PCR with 5'-end labeling generated distinct sequence ladders with a low background, even in specimens containing relatively low copy numbers of HPV. Of 14 cervical neoplasias, 11 cases showed sequence diversity from prototype HPV16, and a total of 22 nucleotide exchanges were detected. Nine of these led to single amino acid exchanges: [Thr5] to [Lys5] in one case and [Asn29] to [Ser29] in eight cases. The [Ser29] E7 was distributed uniformly among invasive carcinomas and precancerous legions, and was also found in a normal cervix. The [Lys5] E7 and [Ser29] E7 had transforming potential similar to the prototype E7 assessed by cooperation with the activated ras gene in rat embryo fibroblasts.

Amino Acid Sequence↗
PubMed1994

Insulin sensitivity, insulin secretion, and glucose effectiveness in subjects with impaired glucose tolerance: a minimal model analysis.

The aim of the present study was to estimate insulin secretion, insulin sensitivity (SI), and glucose effectiveness (SG) in non-obese Japanese subjects with impaired glucose tolerance (IGT). Ten IGT subjects (five men, five women) and 15 normal-tolerance subjects (seven men, eight women) without a family history of diabetes were studied. They underwent a modified frequently sampled intravenous glucose tolerance test (FSIGT); glucose (300 mg/kg body weight) was administered, and insulin (20 mU/kg over 5 minutes) was infused from 20 to 25 minutes after the administration of glucose. SI and SG were estimated by Bergman's minimal model method. No significant difference was observed in body mass index ([BMI] 22.1 +/- 0.8 v 21.1 +/- 0.5 kg/m2), fasting plasma glucose (5.19 +/- 0.18 v 5.07 +/- 0.11 mmol/L), and insulin levels (50.7 +/- 7.3 v 45.2 +/- 4.5 pmol/L) of subjects with IGT and normal controls. The glucose disappearance rate (KG) was significantly lower in subjects with IGT than in normal-tolerance subjects (1.57 +/- 0.20 v 2.09 +/- 0.15%/min, P < .05). Pancreatic insulin secretion expressed as the integrated area of plasma insulin above the basal level during the first 20 minutes was lower in IGT subjects (2,556 +/- 572 pmol/L x min) than in normal-tolerance subjects (4,957 +/- 800 pmol/L x min, P < .05). SI was not statistically different between the two groups (0.84 +/- 0.13 x 10(-4) v 1.14 +/- 0.15 x 10(-4).min-1.pmol/L-1). However, SG was significantly lower in subjects with IGT than in normal controls (0.013 +/- 0.002 v 0.023 +/- 0.002 min-1, P < .01).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗