Search PubMed⌕ Search

Biomedical subjects

K Ohta

Publications and source records attributed to K Ohta.

At least 487 records · Page 27Linked to original sources
PubMed1993

Heterogeneity of mutations in maple syrup urine disease (MSUD): screening and identification of affected E1 alpha and E1 beta subunits of the branched-chain alpha-keto-acid dehydrogenase multienzyme complex.

Maple syrup urine disease (MSUD) is an autosomal recessive disease caused by a deficiency in subunits of the branched-chain alpha-keto-acid dehydrogenase complex (BCKDH). To characterize the mutations present in five patients with MSUD (four classic and one intermediate), three-step analyses were established: (1), identification of the involved subunit by complementation analysis using three different cell lines derived from homozygotes having E1 alpha, E1 beta or the E2 mutant gene; (2), screening for a mutation site in cDNA of the corresponding subunit by RT-PCR-SSCP and (3), mutant analysis by sequencing the amplified cDNA fragment. Four single-base missense mutations, R115W, Q146K [corrected], A209T and I282T, were detected in the E1 alpha subunit. A single-base missense mutation H156R and three frame-shift mutations to generate stop codons downstream, including an 11-bp deletion of the tandem repeat in exon 1, a single-base (T) deletion and a single-base (G) insertion, were identified in the E1 beta subunit gene. All except one (11-bp deletion in E1 beta (Nobukuni, Y., Mitsubuchi, H., Akaboshi, I., Indo, Y., Endo, F., Yoshioka, A. and Matsuda, I. (1991) J. Clin. Invest. 87, 1862-1866)) were novel mutations. The sites of amino-acid substitution were all conserved in other species. Thus, mutations causing MSUD are heterogenous.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
PubMed1993

Growth inhibition of human lung cancer cell lines by interleukin 6 in vitro: a possible role in tumor growth via an autocrine mechanism.

It has been considered that growth of human lung cancer cells, like other malignant cells, is positively and negatively regulated by a variety of growth factors via autocrine as well as paracrine mechanisms. The autocrine mechanism is considered to be important in the autonomy of proliferation of cancer cells. Recently, the role of autocrine growth-inhibiting factors such as transforming growth factor beta attracts special attention for better understanding of growth regulation of malignant cells. Here, we have demonstrated that a multifunctional cytokine interleukin 6 (IL-6) had an inhibitory effect on the proliferation of human non-small cell lung cancer cell lines, as shown by the growth accelerating effect of the specific anti-IL-6 antibody as well as the effect of exogenously added IL-6. Moreover, IL-6 can be expressed and released by human lung cancer cells, and these cells had specific IL-6 receptors on their cell surfaces, suggesting an autocrine mechanism. The growth-inhibitory effect of IL-6 was additive to that of transforming growth factor beta, and could not be neutralized by the addition of anti-transforming growth factor beta antibody. These results suggested that IL-6 may function as another class of autocrine growth-inhibiting factor in the growth regulation of human lung cancer. Relatively lower IL-6 sensitivity of these cells than noncarcinogenic human bronchial epithelial cells also suggested that escape from growth regulation by inhibitory factors such as IL-6 could be involved in lung cancer oncogenesis.

Adenocarcinoma↗
PubMed1993

Nicotinic acetylcholine receptor mRNAs in myasthenic thymuses: association with intrathymic pathogenesis of myasthenia gravis.

Reverse transcription and the polymerase chain reaction showed that the alpha-, beta- and gamma-subunit genes of nicotinic acetylcholine receptor (AChR) in human skeletal muscle were expressed in myasthenia gravis (MG) thymuses (3 thymomas and 7 hyperplastic thymuses). A study of the expression of these subunit mRNAs in a variety of human tissues showed that alpha-subunit mRNA was expressed in human thymus and cerebral cortex, and that all three subunits were expressed only in human child thymus. Our results suggest that a complete AChR similar to extrajunctional (embryonic) muscle AChR is expressed in MG thymuses and that intrathymic AChR might be the primary antigen which induces autoimmune responses to muscle AChR.

Base Sequence↗
PubMed1993

Comparative study of active absorption by the intestine and disposition of anomers of sugar-conjugated compounds.

Active absorption in the intestine and metabolism of the beta- and alpha-anomers of the glucoside and galactoside of p-nitrophenol (p-NP) were studied to find a more suitable prodrug for poorly absorbed drugs. The everted sac technique was used to investigate the intestinal absorption of these glycosides at 250 microM from the mucosal to the serosal side in the rat jejunum. The absorption clearance of p-nitrophenyl alpha-D-glucopyranoside (p-NP alpha glc) (0.271 +/- 0.089 microL/min/cm, mean +/- SE, N = 8) was much lower than that of p-nitrophenyl beta-D-glucopyranoside (p-NP beta glc) (4.45 +/- 0.34 microL/min/cm, mean +/- SE, N = 4) which is actively absorbed by a glucose transport carrier [Mizuma et al., Biochem Pharmacol 43: 2037-2039, 1992]. However, the major constituent appearing on the serosal side was p-NP (aglycone) after absorption of pNP alpha glc, whereas it was p-NP beta glc itself after absorption of p-NP beta glc. The total amount transported to the serosal side after 20 min of p-NP alpha glc absorption, which was similar to that of p-NP beta glc, was significantly decreased in the absence of Na+, indicating the active absorption of p-NP alpha glc by a Na(+)-dependent glucose transport carrier. Perfusion with a mucosal solution of p-NP alpha glc showed that the p-NP concentration on the serosal side (15.8 +/- 1.56 microM, mean +/- SE, N = 3) was significantly (P < 0.05) higher than that on the mucosal side (5.84 +/- 1.24 microM, mean +/- SE, N = 3) at 20 min. This indicated that the p-NP appearing on the serosal side was derived not from absorption of p-NP but from hydrolysis of p-NP alpha glc through the intestinal membrane during absorption. On the other hand, after absorption of p-nitrophenyl beta-D-galactopyranoside (p-NP beta gal), which is actively absorbed by glucose transport carrier, p-NP beta gal itself appeared mostly on the serosal side. However, p-nitrophenyl alpha-D-galactopyranoside (p-NP alpha gal) absorption, which resulted in appearance on the serosal side, was not significantly decreased in the presence of 1 mM phloridzin or in the absence of Na+, indicating that the contribution of the glucose transport carrier to p-NP alpha gal absorption was minimal. The order of the Na(+)-dependent intestinal absorption was p-NP beta glc > p-NP alpha glc > p-NP beta gal > p-NP alpha gal.

Animals↗
PubMed1993

High-level production and secretion of a mouse-human chimeric Fab fragment with specificity to human carcino embryonic antigen in Escherichia coli.

A high-level secretion system for the production of mouse-human chimeric antibody 21B2 (MHC 21B2) Fab fragment specific for human carcino embryonic antigen (hCEA) in Escherichia coli has been constructed. The genes encoding a light chain and an Fd fragment (a variable region and the CH1 domain of a heavy chain) of a mouse-human chimeric antibody were directly fused to the signal peptide of the E. coli ompF gene sequence. E. coli cells containing expression vectors in which each of the two genes are located downstream of a separate tac promoter were able to secrete the light chain and Fd fragment as two of their major cellular proteins. The signal peptides were efficiently removed from the primary products by post-translational processing, although they formed insoluble aggregates, possibly in the periplasm. In high-cell-density culture experiments using a jar fermentor, the amount of light chain and Fd fragment produced was at levels of up to 2.88 g/l and 1.28 g/l culture, respectively. By optimizing the conditions that encourage correct folding, formation of disulphide bonds, and association of the light chain with the Fd fragment, we have established a procedure that can purify, re-fold, and combine aggregated products to electrophoretically homogeneous Fab fragment with a yield of approximately 47%. Fab fragment produced in this manner shows essentially the same antigen-binding activity and specificity to hCEA as the parental mouse antibody 21B2 (MoAb 21B2).

Amino Acid Sequence↗
PubMed1993

Renal effects of manidipine hydrochloride. A new calcium antagonist in hypertensive patients.

The renal effects of manidipine hydrochloride were investigated in ten hospitalised patients with mild-to-moderate essential hypertension. After a one-week placebo period, manidipine was given for 1 week in a dose rising from 5 mg to 10 mg or 20 mg daily to normalise the mean blood pressure measured after 2 h. Blood pressure had decreased from 171/101 to 147/86 mm Hg at the end of manidipine treatment. The pulse rate was unaltered. Renal vascular resistance decreased from 1.90 to 1.33 dyn.s.cm-5/1.48 m2 x 10(4), and renal blood flow and glomerular filtration rate increased from 522 to 662 ml.min-1 x 1.48 m-2 and from 81 to 93 ml.min-1 x 1.48 m-2, respectively, in spite of a fall in renal perfusion pressure. Manidipine reduced the filtration fraction from 0.260 to 0.243, suggesting a preferential reduction in efferent arteriolar resistance. The fractional excretion of sodium and potassium did not change. Manidipine did not produce any significant alteration in plasma renin activity or in the plasma aldosterone concentration. The results indicate that manidipine has favourable renal effects and a concomitant hypotensive action in patients with mild-to-moderate essential hypertension.

Adult↗
PubMed1993

Increased levels of urinary interleukin-6 in Kawasaki disease.

Kawasaki disease (KD) often presents with abnormal urinary findings, such as aseptic pyuria, mild proteinuria and microscopic haematuria. In this study, we measured urinary interleukin-6 (IL-6) by a sensitive sandwich ELISA assay using mouse monoclonal antibodies against recombinant IL-6 to elucidate the role of IL-6 in the pathogenesis of renal lesions in KD. Serum IL-6 levels were increased in acute KD as well as in febrile controls. Importantly, urinary IL-6 levels were consistently elevated in patients with acute KD, but much lower in febrile controls. Urinary IL-6 levels returned steadily to normal during the convalescent phase. In addition to IL-6, urinary levels of N-acetyl-beta-D-glucosaminidase (NAG) and beta 2-microglobulin (beta 2-mg) were also elevated during the acute phase of this disease. Eosinophils and macrophages were identifiable in urinary sediments from these patients. The increased levels of urinary IL-6 in combination with increased NAG and beta 2-mg seemed to suggest the presence of certain renal parenchymal lesions with cellular infiltration during the acute phase of the disease. IL-6 may serve as clinically useful parameter for the detection and monitoring of the renal involvement in KD.

Acetylglucosaminidase↗
PubMed1993

Endothelin-1 is an autocrine/paracrine regulator of porcine granulosa cells.

We studied whether a novel vasoconstrictor, endothelin-1 (ET-1), is synthesized by and released from porcine granulosa cells, and whether ET-1 acts directly on granulosa cells in an autocrine/paracrine fashion. The dilution curve of the conditioned medium from cultured porcine granulosa cells was parallel to a standard curve of ET-1 in RIA. Reverse-phase HPLC of the conditioned media from the granulosa cells revealed a major peak of ET-1-like immunoreactivity (ET-1-LI) coeluting with standard ET-1. ET-1-LI was released from cultured porcine granulosa cells as a function of time. Northern blot analysis demonstrated the expression of mRNA for prepro-ET-1 in the granulosa cells. We demonstrated the presence of ET-1 in the follicular fluid of porcine ovaries, and that the concentration of ET-1 in large follicles was higher than that in small-medium follicles. ET-1 dose-dependently stimulated cell growth and DNA synthesis in porcine granulosa cells. ET-1 inhibited the FSH- and hCG stimulated accumulation of progesterone in porcine granulosa cells in long-term incubation. These findings suggest that ET-1 produced by porcine granulosa cells may function as an autocrine/paracrine growth factor and modulator of steroidogenesis in ovarian granulosa cells.

Animals↗
PubMed1993

Effect of neuropeptide Y on human bronchus and its modulation of neutral endopeptidase.

BACKGROUND: Neuropeptide Y (NPY) and neutral endopeptidase have been shown to regulate the response of airway smooth muscle. However, their interactions have not been studied. METHODS: To determine the role of NPY and endogenous neutral endopeptidase in regulating the effects of NPY, we investigated the effects of NPY and a neutral endopeptidase inhibitor, phosphoramidon, on the contractile response of isolated human bronchial segments to electrical field stimulation (EFS). RESULTS: Concentrations of up to 10(-6) mol/L of NPY did not change resting tension. NPY inhibited the cholinergic contractions induced by EFS at 5 Hz dose-dependently. Phosphoramidon (10(-5) mol/L) enhanced the inhibitory effect of NPY on EFS-induced contractions at 5 Hz. NPY (3 x 10(-7) mol/L) shifted the frequency-response curve for EFS (1 to 50 Hz) to the right and significantly increased the mean logarithm of frequency that produced 50% of maximal contraction (control: 0.54 +/- 0.07 versus NPY: 0.79 +/- 0.09, p < 0.05). Phosphoramidon (10(-5) mol/L) did not affect the frequency response to EFS, but enhanced the inhibitory effect of NPY (3 x 10(-7) mol/L) on frequency response to EFS, resulting in a significant increase in the mean logarithm of frequency that produced 50% of maximal contraction (NPY: 0.79 +/- 0.09 versus NPY plus phosphoramidon: 1.18 +/- 0.15, p < 0.05). NPY (3 x 10(-7) mol/L) did not affect the response of tissues to exogenously applied acetylcholine. CONCLUSIONS: These observations suggest that NPY inhibits the output of acetylcholine from nerve endings and that endogenous neutral endopeptidase modulates the effect of NPY in the human airway by inactivating this peptide.

Acetylcholine↗
PubMed1993

[An outbreak of gastroenteritis due to group C rotavirus in Tokyo].

In April 1991, an outbreak of acute gastroenteritis due to group C rotavirus occurred at an elementary school in Tokyo. Fifty-one (13%) of 393 students became ill. The main clinical symptoms were diarrhea (100%), abdominal pain (68%) and vomiting (56%). No enteropathogenic bacteria were found in the fecal specimens. However, the virus particles morphologically indistinguishable from conventional rotavirus were detected in 6 of 11 fecal specimens by electron microscopy. Immune electron microscopy showed that these virions aggregated with anti-group C rotavirus serum. The RNA pattern of the virus particles involved in this outbreak showed a pattern similar to that of typical group C rotavirus on polyacrylamide gel electrophoresis.

Child↗
PubMed1993

[On the outbreak of Shigellosis in Tokyo, 1992].

In 1992, the number of cases of shigellosis in Tokyo increased remarkably, and a total of 10 separate outbreaks were confirmed. Except for 1 series where the causative strains was Shigella flexneri 3a, the other 9 outbreaks were caused by Shigella sonnei. Of note, 7 outbreaks were familial with less than 5 patients. The remaining 3 outbreaks occurred in a restaurant, a school and the dormitory of an orphanage, in which the organism was detected in 6 to 14 patients. In 4 of the familial outbreaks occurring in distant areas during August, ark shell was presumed to be the vehicle by epidemiological investigations and laboratory examinations. Analysis of isolates such as the antibiotic resistance pattern, plasmid profile, or colicine type in Shigella sonnei was useful in clarifying the commonality of each case and identifying the vehicle or transmission.

Adult↗
PubMed1993

Production of high concentrations of ethanol from inulin by simultaneous saccharification and fermentation using Aspergillus niger and Saccharomyces cerevisiae.

Pure nonhydrolyzed inulin was directly converted to ethanol in a simultaneous saccharification and fermentation process. An inulinase-hyperproducing mutant, Aspergillus niger 817, was grown in a submerged culture at 30 degrees C for 5 days. The inulin-digestive liquid culture (150 ml) was supplemented with 45 g of inulin, 0.45 g of (NH4)2SO4, and 0.15 g of KH2PO4. The medium (pH 5.0) was inoculated with an ethanol-tolerant strain, Saccharomyces cerevisiae 1200, and fermentation was conducted at 30 degrees C. An additional 20 g of inulin was added to the culture after 15 h of fermentation. S. cerevisiae 1200 utilized 99% of the 65 g of inulin during the fermentation, and produced 20.4 and 21.0% (vol/vol) ethanol from chicory and dahlia inulins, respectively, within 3 days of fermentation. The maximum volumetric productivities of ethanol were 6.2 and 6.0 g/liter/h for chicory and dahlia inulins, respectively. The conversion efficiency of inulin to ethanol was 83 to 84% of the theoretical ethanol yield.

Aspergillus niger↗
PubMed1993

Mast cells are essential for the full development of silica-induced pulmonary inflammation: a study with mast cell-deficient mice.

Silicosis provides a good model for chronic interstitial pulmonary inflammation. In order to clarify the role of mast cells in the development of interstitial lung diseases, silica suspension was transnasally administered to mast cell-deficient mice (WBB6F1-W/Wv) and their mast cell-intact littermates (WBB6F1(-)+/+) as well as to normal mice (C57BL/6). Histologic examinations and analyses of bronchoalveolar lavage fluid (BALF) components indicated that silica instillation induces less severe lung lesions in mast cell-deficient mice than in mast cell-intact mice. BALF neutrophilia was prominent in mast cell-intact mice, but mast cell-deficient mice developed significantly milder BALF neutrophilia. An increase in the number of lung mast cells was observed in mast cell-intact mice. To further prove the involvement of mast cells, bone marrow-derived cultured mast cells from +/+ mice were adoptively transferred to mast cell-deficient mice. These mast cell-reconstituted mice developed more severe pulmonary lesions than did the mast cell-deficient mice; the severity of the lesions was similar to that in mast cell-intact mice. In addition, BALF neutrophilia was elicited by mast cell reconstitution. A significant number of mast cells was found in the lungs of mast cell-reconstituted mice when silica was administered. These results suggest the involvement of mast cells in the development of silicosis and implicate interactions between mast cells and neutrophils in the pathogenesis of this disorder.

Animals↗