Biomedical subjects

M Shingu

Publications and source records attributed to M Shingu.

At least 19 recordsLinked to original sources

PubMed1992

Activation of complement in normal serum by hydrogen peroxide and hydrogen peroxide-related oxygen radicals produced by activated neutrophils.

Neutrophils activated by soluble particulate stimuli generate superoxide anion and subsequently form hydrogen peroxide and other oxygen radicals. The effect of hydrogen peroxide on the complement system in normal serum was investigated. Treatment of normal serum with hydrogen peroxide resulted in a diminution of the haemolytic activity of the total and alternative complement pathways and the haemolytic titres of C3 and C5 but not of C2, in normal serum. These decreases in complement activity depended on the concentration of hydrogen peroxide added to the serum. Immunoelectrophoretic analysis of hydrogen peroxide-treated serum showed that C3 and C5 proteins were activated. Complement degradation products C3a and C5a were produced in normal serum treated with hydrogen peroxide, and 20 mM EDTA abolished C3a and C5a production in hydrogen peroxide-treated serum but 20 mM Mg-EGTA did not. Catalase completely abolished and dimethylsulphoxide and D-mannitol, hydroxyl radical scavengers, partially inhibited the hydrogen peroxide-mediated complement activation. Hypochlorite, incubated with normal serum, significantly inhibited serum haemolytic activity, and sodium thiosulphate, a reducing agent, abolished the effect of hypochlorite. Normal serum incubated with activated neutrophils showed neutrophil chemotactic activity and decreased serum haemolytic activity, and the addition of catalase or methionine (5 mM) completely abolished the effects of activated neutrophils. These results suggest that hydrogen peroxide activates complement via an alternative pathway of complement activation and that hydroxyl radicals and other hydrogen peroxide-related species such as hypochlorite are most likely involved in hydrogen peroxide-mediated complement activation. Complement activation by oxygen radicals produced by activated neutrophils may be one of the mechanisms by which complement is activated in human immune complex diseases.

Complement Activation↗

PubMed1992

[Detection and serotyping of HRVs collected from children with acute gastroenteritis in winter of 1986 to 1991].

Fecal samples from sporadic acute gastroenteritis patients who visited at a pediatrician during five consecutive winter seasons in Kurume City located at north part of Kyushu district. Fecal samples were subjected to detection of rotavirus and other viruses and study of yearly changes in serotype of rotavirus. The rate of group A rotavirus positives in third winter and other consecutive winters were 12.5%, 40 to 65%, respectively. 60% specimens were successfully serotyped: 58% was serotype 1.22% was serotype 3.6% was serotype 2, and 14% was serotype 4. The dominant serotype in first, fourth and fifth winter was type 1. However in second winter each serotype of rotavirus was almost equally detected. During five consecutive winters six patients visited at the same pediatrician twice more than two months interval. They all had acute gastrointestinal symptoms. In six patients rotavirus was detective just once.

Acute Disease↗

PubMed1992

Pattern recognition of continuity of discrete data points by scale changed plots.

Graphs composed of a large number of data points plotted on an appropriate scale enables us to pattern recognize a specific continuous curve. This principle was used to follow signals from proton nuclear magnetic resonance as a function of pH in a large biomolecule, staphylococcal nuclease. The analysis was automated by auto peak-picking routines and by transferring the results to a graphic program, Graphic Operating System (Roome & Peterson, 1985). Expansion and contraction of the scale made it possible to differentiate specific tyrosine and histidine titration curves. This type of analysis is applicable for assignment of specific curves from a large number of data points which change as a function of perturbations.

Magnetic Resonance Spectroscopy↗

PubMed1992

Construction of an expression vector for human papillomavirus type 16 E7-glutathione S transferase fusion protein.

An expression vector for human papillomavirus type 16 (HPV 16) E7-glutathione S transferase fusion protein was constructed. The E7 gene was confirmed by southern blot analysis with a digoxygenin 11-dUTP labeled and simultaneously amplified E7 DNA probe by polymerase chain reaction (PCR). The fusion protein was expressed in E. coli, recovered as inclusion bodies, and was confirmed by western blot analysis using a monoclonal antibody against HPV 16-E7 protein. A polyclonal antibody against the HPV 16-E7 protein was raised in the serum of mice hyper-immuned with the fusion protein. This antibody will be available for screening of patients with cervical cancer.

Base Sequence↗

PubMed1992

[Rapid detection of mycobacteria in sputa using media supplemented with culture filtrate of Gemella haemolysans].

The effect of the culture filtrate Gemella haemolysans in enhancing mycobacterial growth has been previously demonstrated. In the present studies, an attempt was made to confirm whether the addition of the filtrate into the medium would be an effective method to promote the rapid detection of mycobacteria in sputum specimens of patients. One-hundred and one sputum specimens pretreated with NaOH were inoculated with various media (Dubos, Dubos-agar, 1% Ogawa, Kudo PD, and Middlebrook 7H9) supplemented with the culture filtrate of Gemella haemolysans grown in blood-BHI or blood-HEM at the dilutions of 1/32 or 1/64. Addition of the filtrate reduced the amount of time required to detect mycobacterial growth (Mycobacterium tuberculosis, M. avium complex, and M. kansassi) by an average of 60-70%. The media containing the filtrate formed significantly larger numbers of colonies compared with the control media, and those colonies inoculated developed more rapidly in size. The findings clearly indicated that the culture filtrate promotes the effective growth of mycobacteria which cannot or only slowly grows in ordinary media. Also indicated was that the addition of the culture filtrate of Gemella haemolysans into media provides as a useful tool to allow the rapid diagnosis of mycobacterial diseases.

Bacteriological Techniques↗

PubMed1992

The possible role of endothelial cells in hypertensive disorders during pregnancy.

OBJECTIVE: To clarify the role of endothelial cells in pregnancy-related hypertensive disorders, we studied the cytotoxic effect of sera from normal pregnant women and from gravidas with various hypertensive complications of pregnancy. METHODS: We obtained serum samples from 84 Japanese women: 17 with preeclampsia, ten with gestational hypertension, six with chronic hypertension, five with chronic hypertension with superimposed preeclampsia, 21 normal gravidas, and 25 healthy nonpregnant women. Endothelial cell injury was measured by the release of radiolabeled chromium from the cells into the culture medium. RESULTS: The mean (+/- standard error of the mean) values of chromium 51 release in preeclampsia, gestational hypertension, chronic hypertension, chronic hypertension with superimposed preeclampsia, normal pregnancy, and healthy nonpregnant women were: 21.9 +/- 2.1, 10.0 +/- 2.0, 9.2 +/- 2.3, 12.9 +/- 0.8, 8.4 +/- 1.4, and 7.3 +/- 1.6%, respectively. Normal pregnant and nonpregnant subjects did not differ with respect to endothelial cell injury. Sera from women with preeclampsia demonstrated significantly greater endothelial cell injury than did sera from normal gravidas. Subjects with the three other categories of hypertensive disorders did not differ significantly from normal gravidas. CONCLUSION: Preeclampsia is characterized by the presence of a serum factor cytotoxic to endothelial cells. Therefore, the mechanism responsible for the increase in blood pressure differs between women with preeclampsia and those with other hypertensive disorders in pregnancy.

Adult↗

PubMed1991

Restricted diversity of the variable region nucleotide sequences of the heavy and light chains of a human rheumatoid factor.

The complete nucleotide sequences of the variable region genes of the heavy and light polypeptide chains of a human monoclonal rheumatoid factor (RF) produced from a human-mouse heterohybridoma were determined. The antibody, designated YES8c, contained V kappa III, J kappa 2, VH1, JH4, and a D gene segment of 9 amino acids. The nucleotide sequences and the deduced amino acid sequences of the light chain variable region were remarkably homologous (97-98%) to previously described RF of the Wa idiotypic family (PAY, GLO, CUR, FLO, and GAR) and to that of a V kappa III germline gene (Humkv325). The YES8c heavy chain variable region gene was most closely related to the VH1 gene of the restricted human fetal repertoire, designated 51p1, and also to 3 rearranged VH1 genes that were recently isolated from patients with chronic lymphocytic leukemia. These results suggest that variable region genes of RFs are highly conserved and that YES8c VH, as well as V kappa, may be identical to heavy and light chains expressed during early B cell development.

Amino Acid Sequence↗

PubMed1991

Effects of recombinant human IL-1 beta on production of prostaglandin E2, leukotriene B4, NAG, and superoxide by human synovial cells and chondrocytes.

The effects of recombinant human IL-1 beta on the production of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), N-acetyl-beta-D-glucosaminidase (NAG), and superoxide by synovial cells and chondrocytes derived from osteoarthritis patients were determined. IL-1 beta markedly enhanced PGE2 production in chondrocytes and, to the lesser extent, in synovial cells. Synovial cells and chondrocytes spontaneously released LTB4 into culture medium and IL-1 beta significantly inhibited LTB4 production by these cells. IL-1 beta significantly suppressed the release of NAG and superoxide by synovial cells, whereas it significantly enhanced the production of NAG and superoxide by chondrocytes. Production of intracellular superoxide dismutase by synovial cells was significantly enhanced on incubation with IL-1 beta, but that of chondrocytes was not altered. IL-6, unlike IL-1 beta, significantly suppressed the production of NAG and superoxide by synovial cells and chondrocytes. These results suggest that IL-1 has differing effects on the release of mediators by synovial cells and chondrocytes and that these cells also vary in their responses to IL-1 beta and IL-6.

Acetylglucosaminidase↗

PubMed1991

Relationship between leukotriene B4 and immunological parameters in rheumatoid synovial fluids.

Leukotriene B4 (LTB4) was measured in synovial fluid from 20 patients with rheumatoid arthritis and 15 patients with osteoarthritis. The level of LTB4 was significantly higher in synovial fluid from rheumatoid arthritis patients as compared with synovial fluid from osteoarthritis patients. LTB4 levels also significantly correlated with cell numbers, rheumatoid factor, and immune complexes in synovial fluid from rheumatoid arthritis patients. There was an inverse correlation between LTB4 levels and complement components. The high-pressure liquid chromatography peak of immunoreactivity extracted from the synovial fluid occurred at a retention volume identical to that of authentic LTB4. These results suggest that the increased level of this mediator in synovial fluid may contribute to perpetuation of inflammation and tissue destruction in rheumatoid arthritis.

Adult↗

PubMed1991

Multicentric reticulohistiocytosis associated with subclinical Sjögren's syndrome.

A case of multicentric reticulohistiocytosis in a 60-year-old Japanese woman associated with subclinical Sjögren's syndrome is presented. The clinical features along with the light microscopic studies are commented. Partial improvement of the skin lesions and tendon sheath swelling was achieved after treatment with cyclophosphamide but the patient's general condition remained unchanged. Taking the rarity of multicentric reticulohistiocytosis into account, coexistence of these two conditions in the present and previously reported cases suggests that an autoimmune mechanism may play a part in the pathogenesis of multicentric reticulohistiocytosis.

Cyclophosphamide↗

PubMed1991

Recombinant human IL-1 beta and TNF-alpha stimulate production of IL-1 alpha and IL-1 beta by vascular smooth muscle cells and IL-1 alpha by vascular endothelial cells.

Vascular endothelial cells (EC) produced IL-1 alpha but not IL-1 beta into extracellular fluids. Vascular smooth muscle cells (SMC), on the other hand, produced both IL-1 alpha and IL-1 beta, and IL-1 beta produced was much higher than IL-1 alpha. The addition of recombinant human IL-1 beta or recombinant human TNF-alpha significantly enhanced IL-1 alpha production in EC, and IL-1 alpha and IL-1 beta production in SMC. IL-1 beta release was not observed even when EC were stimulated with TNF-alpha. These results suggest that the species of released form of IL-1 are different in different cell types and that cytokines enhance IL-1 alpha and IL-1 beta production in SMC and IL-1 alpha production in EC.

Cells, Cultured↗

PubMed1991

Densitometric determination of human papillomavirus DNA quantities by chromato-scanning in the fluorescence mode.

The quantitation of human papillomavirus DNA isolated from warts by chromato-scanning (fluorescence mode) photographs of ethidium bromide-stained agarose gels is described. Excitation at 200 nm (with a cutoff filter at 400 nm) generates fluorescence from the white portion of the printing paper. The fluorescent intensity correlated with the quantities of DNA in the band of interest. The amounts of DNA were determined using calibration curves of approximately the same size as lambda phage DNA fragments. This general method of quantification is applicable to photographs of other types of polynucleotides capable of being separated and stained in a gel medium.

Bacteriophage lambda↗

PubMed1991

Therapeutic effects of bovine enterovirus infection on rabbits with experimentally induced adult T cell leukaemia.

A bovine enterovirus, MZ-468, showed cytopathic effects on cell line F-647a, which was established by coculture of human T cell lymphotropic virus type 1-transformed MT-2 cells and X-irradiated rabbit lymphocytes. Microcalorimetric assay showed that residual, viable, MZ-468-infected F-647a cells produced less heat than non-infected cells. The therapeutic effects of MZ-468 infection were examined in rabbits in which adult T cell-like leukaemia (ATL) had been induced by inoculation of F-647a cells (1 x 10(8) cells). Six newborn rabbits were separated into three groups: group A was inoculated with F-647a cells only; group B was treated with MZ-468 at the time of inoculation with cells; group C was treated with the same amount of virus 24 h after the inoculation with cells and then once every 4 days. Both of the animals in group A and one in group C died 10 and 11 days, and 22 days, respectively, after the inoculation with cells. Both rabbits in group B and one in group C survived for more than 4 months. The rabbits that died were examined pathologically; leukaemic infiltrations were found in the lungs of the group B rabbits, and in the lungs, spleens and livers of both group A rabbits. Two identical experiments produced almost the same findings. These results suggest that bovine enterovirus might be used clinically to prolong the life-span of ATL patients.

Animals↗

PubMed1991

Improvement of the plaque technique for human rotaviruses: effect of fetal bovine serum, acetyltrypsin and agar.

Dilution of viruses, which were previously treated with 10 micrograms acetyltrypsin/ml, in MEM supplemented with 2.5% fetal bovine serum caused little damage of cell monolayers resulting in the formation of discrete plaques. When agar and acetyltrypsin were added to the overlay medium at the concentrations of 0.8% and 5 micrograms/ml, respectively, clear plaques were induced. Linear relationships were obtained between virus concentration and the number of plaques, thus the procedure described in this report might be used for reliable plaque assay of rotaviruses. Moreover, these conditions for plaque formation were successful for estimating isolated viruses, as well as standard strains.

Agar↗

PubMed1991

A pH-dependent conformational change of staphylococcal nuclease: hydrogen-1 two-dimensional nuclear Overhauser spectroscopy studies.

Two-dimensional nuclear magnetic resonance (2d-NMR) dipolar correlation spectroscopy (NOESY) for the calcium binding protein, staphylococcal nuclease, was used to study the conformations in solution as a function of pH. A series of 2d-NOESY spectra of the nuclease H124L-Ca(++)-thymidine 3'5'-bisphosphate ternary complex demonstrated significant pH dependent changes: NOE cross peaks between 46His delta H and an aliphatic proton at 2.66 ppm appeared at pH 10.2, but not at pH 5.5. Some other NOE cross peaks between aromatic and aliphatic protons displayed chemical shift changes in omega 2 or both dimensions (omega 1, omega 2). These results indicated that a local rearrangement accompanies the ionization of the functional groups and has a role in enzyme function.

Hydrogen↗

PubMed1991

Restriction endonuclease analysis of varicella-zoster virus DNAs.

The DNAs of 14 strains of varicella-zoster virus (VZV) isolated from epidemiologically unrelated individuals were analyzed by restriction endonuclease cleavage and agarose gel electrophoresis. Small but distinct differences in restriction patterns between the isolates were observed when the DNAs were digested with each of four enzymes examined (EcoRI, SalI, PstI, and HindIII). Six out of 14 isolates (43%) were lacking the PstI cleavage site between K and N fragments; one isolate was lacking the HindIII cleavage site between M and N fragments. No changes were observed in restriction patterns in DNA of one of the strains after 33 passages in cultured cells.

DNA, Viral↗

PubMed1991

Correlation between DNA quantities in agarose gel and chromatographic areas of photographed DNA bands.

To quantitate DNA from photographs, correlations between chromato-scanned (in the fluorescence mode) and virtual areas from white DNA bands on photographs were examined. A linear correlation between the two measurements with coefficients higher than 0.89 was observed. The dynamic range of the fluorescence detection yielded good linearity for a wide range of amounts of DNA (microgram-ng) on the photograph.

Chromatography↗

PubMed1990

Human papillomavirus DNA from warts for typing by endonuclease restriction patterns: purification by alkaline plasmid methods.

The alkaline plasmid DNA extraction method of Birnboim and Doly was applied for the isolation of human papillomavirus (HPV) from warts. Tissue from common and plantar warts was digested with proteinase K, and the extrachromosomal circular covalently-closed form of HPV-DNA was rapidly extracted by alkaline sodium dodecyl sulphate and phenol-chloroform treatment. Recovery of HPV-DNA from the tissue was sufficient for determination of endonuclease restriction patterns by agarose gel electrophoresis.

Adolescent↗