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Biomedical subjects

F Ota

Publications and source records attributed to F Ota.

At least 19 recordsLinked to original sources

Alterations in serum levels of trace elements in tuberculosis and HIV infections.

OBJECTIVE: To evaluate serum concentrations of trace elements in tuberculosis (TB) patients with or with out human immunodeficiency virus (HIV) coinfection before and after anti-TB chemotherapy. SUBJECTS: A total of 155 TB patients, 74 of which were coinfected with HIV, and 31 healthy controls from Gondar, Ethiopia. METHODS: Serum levels of copper, zinc, selenium and iron were determined using an inductively coupled plasma mass spectrometer from all subjects at baseline and from 44 TB patients (22 with HIV coinfection) at the end of an intensive phase of anti-TB chemotherapy. RESULTS: Compared with the control group, the concentrations of iron, zinc and selenium were significantly lower (P<0.05) while that of copper and copper/zinc ratio was significantly higher (P<0.05) in the serum of TB patients. TB patients with HIV coinfection had significantly lower serum zinc and selenium concentrations and significantly higher copper/zinc ratio compared to that in TB patients without HIV coinfection (P<0.05). The serum concentration of zinc had significantly increased at the end of intensive phase of anti-TB chemotherapy in patients without HIV coinfection (P<0.05). An increase in serum selenium level was observed in TB patients with or without HIV coinfection after therapy. On the contrary, serum copper concentration and copper/zinc ratio declined significantly after anti-TB chemotherapy irrespective of HIV serostatus (P<0.05). CONCLUSIONS: The results indicate that TB patients have altered profile of trace elements in their sera. This warrants the need for further investigations so that strategies for trace elements supplementation can be planned in addition to their potential as diagnostic parameters in monitoring responses to anti-TB chemotherapy.

Adult↗

Serological diversity demonstrable by a set of monoclonal antibodies to eight serotypes of the mutans streptococci.

A set of monoclonal antibodies were prepared by the conventional cell fusion of myeloma cells (SP2/0-Ag14) with spleen cells from BALB/c mice immunised with whole cells of a strain of mutans streptococci. Their specificities were examined against 35 reference strains of mutans streptococci, 34 reference strains of other oral streptococci and 8 reference strains of other microorganisms often inhabiting the oral cavity. Specificity was examined by enzyme immunoassay using whole cells. A total of 52 strains, consisting of 19 strains isolated in Japan, 19 strains isolated in Italy and 14 strains isolated in England, were characterised by conventional physiological and biochemical tests and then serotyped by the use of 8 monoclonal antibodies with different specificities. They were also confirmed by guanine-plus-cytosine contents of their nucleic acid and DNA-DNA hybridisation test. The results indicated that all monoclonal antibodies are useful for identification of 8 serotypes of the mutans streptococci responsible for dental caries. They also suggest the existence of more serological varieties among mutans species.

Antibodies, Bacterial↗

Serum IgE levels of tuberculosis patients in a tropical setup with high prevalence of HIV and intestinal parasitoses.

Tuberculosis remains a major health problem worldwide in the era of HIV/AIDS. Co-infection with intestinal parasites has been suggested to worsen the outcome of infection by polarizing the immune response towards Th2. This study investigated serum IgE levels of 241 tuberculosis patients and compared the IgE profiles in the tuberculosis patients either with or without intestinal helminthic infection and/or HIV infection. The serum levels of IgE in tuberculosis patients before initiation of antimycobacterial chemotherapy were found to be 1722 +/- 1290 IU/ml (Mean +/- SD) in HIV seronegatives and 2366 +/- 1849 IU/ml in HIV seropositives. Further, the IgE level was significantly higher in patients coinfected with intestinal helminthes and HIV compared to those infected with helminthes or without coinfection (P < 0.05). Anti-tuberculosis chemotherapy significantly reduced serum IgE levels in HIV seronegative tuberculosis patients (P < 0.05). These findings might indicate an active role of therapy in shifting the immune response towards Th1 which is crucial for prognosis in tuberculosis patients.

Adult↗

Effect of removable orthodontic appliances on oral colonisation by mutans streptococci in children.

Little is known about the effect of removable orthodontic appliances on oral colonisation by mutans streptococci (MS). In the present study, the frequency of isolation and serotype distribution of MS were evaluated in two groups of children, one undergoing therapy with removable appliances and the other not subjected to any kind of orthodontic treatment, respectively. Streptococci isolated from dental plaque samples from both groups of children were identified as mutans streptococci on the basis of their morphological and biochemical properties and were then serotyped in an enzyme immuno-assay using monoclonal antibodies. The number of subjects harbouring MS in their dental plaque was statistically higher in the group of orthodontic children without caries experience (CF) in comparison with CF children of the control group (10/12, 83.3% vs. 15/44, 34%). No clear difference was observed in the distribution of the different MS serotypes between the experimental and control group: S. mutans c,f serotype was the most frequently isolated in both groups of children followed by S. mutans serotype e and S. sobrinus serotype g. Such results suggest that the use of removable appliances may lead to the creation of new retentive areas and surfaces, which favour the local adherence and growth of MS. The data obtained stress the importance of a careful monitoring of patients treated orthodontically for risk of caries development.

Case-Control Studies↗

Comparison of ovarian follicular activity during treatment with a monthly injectable contraceptive and a low-dose oral contraceptive.

Ovarian follicular development occurs during treatment with combined and progestin-only oral contraceptive (OC) pills and progestin-containing subdermal implants, and can be associated with the development of persistent functional cysts that may require surgical removal. Lunelle is a once-a-month injectable contraceptive containing estradiol cypionate 5 mg and medroxyprogesterone acetate 25 mg. A randomized, comparative study was undertaken to compare the effect on ovarian follicular activity associated with use of Lunelle and a low-dose OC. A total of 30 ovulatory subjects were randomly assigned to receive two cycles of treatment with either an OC containing ethinyl estradiol 20 microg and 0.1 mg levonorgestrel or Lunelle. During the second cycle of treatment, pelvic sonography was performed every 4 days, at which time the maximum follicle diameter was measured. Study end points were the presence of follicles >/=10, 20, and 30 mm. In all, 13 of 15 subjects in the OC group and 14 of 15 in the Lunelle group completed the study. Follicles measuring >/=10 mm were present in 11 of 13 (84.6%) in the OC users and in four of 14 (28.6%) in the Lunelle users (p <0.05). In the OC group, six of 13 subjects (46.1%) developed follicles >/=20 mm, and one of 13 (7.7%) developed follicles >/=30 mm. No subjects in the Lunelle group developed a follicle >/=20 mm in diameter. This study indicates that Lunelle is associated with a significantly lower incidence of ovarian follicular development compared to that of an OC containing 20 microg ethinyl estradiol and 0.1 mg levonorgestrel.

Adult↗

Effects of pH variation and NaCl on in vitro digestibility of cow's milk proteins in commercially available infant formulas.

Allergenicity and antigenicity of food proteins are generally dependent on their heat-stability and resistance to digestive enzymes. Using the methods of SDS-PAGE and immunoblots, we assessed peptic-digestibility of major milk proteins (casein, beta-lactoglobulin and alpha-lactalbumin) in commercially available infant formulas in acidic pH range (pH 1.5 to 4.0), and we also investigated the effect of NaCl on peptic-digestibility of cow's milk proteins at pH 2.0. The profile of peptic-digestion of casein was similar at pH range from 1.5 to 3.5 where the intact protein of casein rapidly disappeared, whereas slow digestibility of the protein was found at pH 4.0. Beta-lactoglobulin and alpha-lactalbumin were digested at pH range from 1.5 to 2.5 and were almost entirely resistant to peptic digestion at over pH 3.0. In the presence of NaCl at pH 2.0, casein showed a good digestibility by pepsin similar to that in the absence of NaCl, in contrast to beta-lactoglobulin and alpha-lactalbumin both having their peptic-digestibility decreased in the presence of 0.2 M NaCl. As the state of the stomach in new born infants shows low amounts of secretary pepsin and out of optimum pH of peptic activity, low digestibility of beta-lactoglobulin and alpha-lactalbumin in cow's milk based infant formulas at over pH 3.0 is supposed to be responsible for their allergenicity.

Animals↗

Characterization of serotype II polysaccharide antigen of group E streptococci using a monoclonal antibody.

A Monoclonal antibody (MAb II-T) specific for serotypes II and V Group E streptococci (GES) was prepared by fusing myeloma cells with spleen cells of mice immunized with whole cells of a serotype II strain. MAb II-T reacted in an enzyme immunoassay (EIA) with whole cells of both serotypes and reacted in gel diffusion test with autoclaved-saline extraction of serotypes II and V. The extract was purified by DEAE-Sephadex A-25, followed by treatment with proteinase K, and further by chromatography with a Sephadex G-200 column. The purified polysaccharide (PS) antigen contained 98.6% carbohydrate and 1.4% protein, but no detectable phosphorus. In hapten inhibition tests using various sugars, D-mannosamine markedly inhibited the precipitin reaction. These results indicated that the antigenic determinant might have a structure similar to D-mannosamine.

Antibodies, Bacterial↗

Quantitation by enzyme immunoassay of spirosin from Lactobacillus reuteri and Escherichia coli.

Three EIA methods (Direct, Indirect and Sandwich EIA) were studied to quantify spirosin in Lactobacillus reuteri and Escherichia coli cultured under various conditions in an attempt to get some insight into the function of spirosome. Both Direct and Indirect EIA were suited well for the quantitation of L. reuteri spirosin while Direct EIA was appropriate for spirosin of E. coli. Sandwich EIA could not be applied successfully in either case. By use of these methods, the amounts of spirosin produced by E. coli were determined to be 1.4, 36.2 and 46.5 micrograms per mg protein of the cell lysate under aerobic, standing and anaerobic culture conditions, respectively. Since the production profile of spirosin coincided entirely with that of alcohol dehydrogenase, these findings supported the identity of spirosin to alcohol dehydrogenase in E. coli. In the same way, L. reuteri spirosin was quantified to be 73.5 and 65.4 micrograms/ mg protein of the lysate in standing and anaerobic culture, respectively. The production pattern of spirosin did not parallel that of alcohol dehydrogenase among three strains of L. reuteri, suggesting that spirosin might not be identical to alcohol dehydrogenase.

Aerobiosis↗

Possible role for a polysaccharide antigen shared between Streptococcus pyogenes and S. mutans in the pathogenesis of poststreptococcal glomerulonephritis.

Streptococcus mutans has been shown to share a polysaccharide (PS) antigen with S. pyogenes strains isolated from patients with acute poststreptococcal glomerulonephritis (PSGN), using a monoclonal antibody f-77 reactive with the PS. To investigate the pathogenetic role of the shared PS in PSNG, experimental nephritis was induced in animals. Rats were immunized thrice with heat-killed cells of S. mutans or S. pyogenes, followed by an intravenous injection of live cells of S. pyogenes. Histologic examination showed that both animal groups had comparable degrees of diffuse proliferative nephritis characterized by immune deposits. The shared PS antigen was detected in glomeruli of all nephritic rats by immunofluorescence using monoclonal antibody f-77. Furthermore, all nephritic rats had an elevated antibody titer to the shared PS antigen. These results suggest that prior sensitization (infections such as dental caries) to S. mutans modulates immune responses to subsequent S. pyogenes infections and induces immune-complex disease (PSGN) through the shared PS antigen.

Acute Disease↗

Immunochemical analysis and localisation of a cell wall antigen of Streptococcus rattus with a specific monoclonal antibody.

A monoclonal antibody specific to a cell wall antigen of Streptococcus rattus (S. rattus) was prepared after cell fusion of mouse myeloma cells to the spleen cells of mice immunized with whole cells of strain FA1. The monoclonal antibody reacted with a crude and purified polysaccharide antigen of S. rattus in precipitin reactions as well as in a solid phase enzyme assay (EIA). It also reacted in the EIA with whole cells of S. rattus strains, but did not react with any other species of the mutans group or of other oral and streptococcal species. Moreover, among many haptenic sugars and their derivatives tested, D-galactosamine and D-glucosamine strongly inhibited the reaction in a competitive enzyme immunoassay using the purified antigen and whole cells, indicating that the type-specific antigenic determinant of the organism has a structure similar to the amino group of the sugar molecules. Furthermore, immunoelectron microscopy revealed that the antigen epitopes formed an irregular fibrous structure over the entire surface of individual cells.

Animals↗

Chromatographic study of spirosin by use of monoclonal antibodies to spirosin of Yersinia enterocolitica.

Two monoclonal antibodies (MAbs) reacting with spirosins from Enterobacteriaceae were obtained in a course of screening MAbs to spirosin from Yersinia enterocolitica SYT-11-72 (YE72). The antibodies were designated MAbs-S44 and S50. They were IgG2b and IgG2a, respectively, both with kappa light chains. On Western blotting after limited proteolysis of YE72 spirosin with Staphylococcus aureus V8 protease, they reacted markedly with peptide fragments of 27 and 35 kDa, suggesting the presence of an antigenic determinant on the fragments. When supernatant cell lysate from Escherichia coli K12 was chromatographed on DEAE-cellulose and Sepharose CL-6B columns successively, a 96-kDa protein with alcohol dehydrogenase (ADH) activity was always associated with reactivity to MAb-S50. These findings combined with N-terminal amino acid sequences clearly indicate the identity of spirosin to ADH in E. coli.

Alcohol Dehydrogenase↗

Detection of Streptococcus mutans by PCR amplification of spaP gene.

Synthetic oligonucleotide primers were used in the polymerase chain reaction (PCR) to amplify a sequence of the spaP gene, which encodes the surface protein antigen I/II of Streptococcus mutans. A DNA fragment of c. 192 bp was amplified from lysed S. mutans cells or isolated DNA. With S. mutans cells, the lower limit of detection was 4-40 cfu. With these primers, 13 reference and 50 clinical strains of S. mutans were identified. Amplification of the 192-bp product was not demonstrated when 41 strains of other streptococcal and non-streptococcal species were tested. The spaP gene PCR has potential for the rapid diagnosis of S. mutans infections.

Adult↗

Monoclonal antibodies to spirosin of Yersinia enterocolitica and analysis of the localization of spirosome by use of them.

Two hybridomas producing monoclonal antibodies (MAbs) were prepared by fusing myeloma cells (Sp2/0-Ag14) with mouse spleen cells immunized with purified spirosin from Yersinia enterocolitica SYT-11-72 (YE72). The antibodies produced by them were designated MAbs-S5 and S27. They were IgG2a and IgG1, respectively, both with kappa light chains. MAbs-S5 and S27 reacted specifically with spirosin from YE72. On Western blotting after limited proteolysis with Staphylococcus aureus V8 protease, YE72 spirosin revealed peptide fragments of 35 and 37 kDa reacting markedly with MAb-S5, which suggested the presence of an antigenic determinant on these fragments. By cellular fractionation of YE72 and subsequent EIA and Western blot analysis, spirosome was shown to be present in the cytoplasm of YE72.

Amino Acid Sequence↗

Immunochemical study of polysaccharide antigen in Streptococcus sobrinus and Streptococcus downei with a cross-reactive monoclonal antibody.

A monoclonal antibody (mAb h-448) was prepared after cell fusion of mouse myeloma cells (SP2/0-Ag-14) to the spleen cells of mice immunised with serotype h strain (MF25) of Streptococcus downei. The antibody (IgM class) reacted in enzyme immunoassay only with whole cells as well as purified polysaccharide (PS) antigen of Streptococcus sobrinus (types d and g) and Streptococcus downei (serotype h), but not with cells or purified PS antigen from any other serotypes of the mutans group of streptococci. mAb h-448 also quantitatively precipitated in solution with the purified antigens. Competitive hapten inhibition tests demonstrated that beta-methylgalactopyranoside inhibited the reaction most strongly. Although rhamnose also showed a substantial inhibitory effect, the results of this study indicate that the antigenic determinant of the PS antigen has a structure similar to the beta-methylgalactopyranoside molecule.

Antibodies, Bacterial↗

First characterization in Italy of clinical isolates of mutans streptococci by using specific monoclonal antibodies.

The aim of this investigation was to gain further insight into the prevalence of different serotypes of mutans streptococci in the Italian population by using specific monoclonal antibodies in an enzyme immunoassay. Isolates from dental plaque samples, collected from an adult population living in Pisa (Italy), were identified as mutans streptococci on the basis of their morphological and biochemical properties, and were then serotyped. The results show that 77.5% of the strains isolated belonged to serotype c or f (i.e., S. mutans), 15.9% were serotype e (i.e., S. mutans) and only two strains (1.4%) belonged to serotype g (i.e., S. sobrinus). These data are partially in agreement with other studies in Europe and in the U.S.A. The distribution pattern of the various serotypes turned out to be substantially similar among the different groups of patients, subdivided on the basis of their caries status, indicating that none of the serotypes was particularly associated with dental caries.

Adolescent↗

Epidemiological survey of Streptococcus mutans in a group of adult patients living in Pisa (Italy).

An epidemiological investigation was carried out to evaluate the prevalence of Streptococcus mutans in a group of 134 adult patients. Markedly higher frequency of isolation was observed in caries-active subjects than in caries-inactive or caries-free subjects, indicating a significant association between the prevalence of the microorganism and the caries status. Moreover, the presence of the microorganism appeared to have a significant association with the extent of caries experience evaluated by the DMF score. These findings are in agreement with those reported previously for school children in other areas of Italy. Isolation of S. mutans was compared among patients groups with different caries activity in relation to culture times of dental plaque samples in a transport medium (Colorimetric Broth Medium). S. mutans was most frequently isolated from caries-active subjects when the medium was incubated for 48 h after inoculation with dental plaque samples.

Adolescent↗