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Biomedical subjects

H Isogai

Publications and source records attributed to H Isogai.

At least 55 records · Page 3Linked to original sources

Heat shock protein produced by Helicobacter pylori.

The cells of Helicobacter pylori were suspended in the medium containing 35S-methionine. After a heat shock of the cells at 42 C for 5, 10, and 30 min, the production of proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Out of many proteins produced by the cells, only 66 kDa protein production was dramatically increased by heat treatment. The N-terminal amino acid sequence of 66 kDa protein was quite similar to that of 62 kDa and 54 kDa proteins previously suggested as heat shock protein (HSP) of H. pylori based on the reaction with polyclonal and monoclonal antibodies against HSP 60 family proteins produced by other bacteria. Therefore, it was concluded that H. pylori produces the 66 kDa protein as its major heat shock protein which belongs to HSP 60 family.

Amino Acid Sequence↗

Complement-mediated killing of Borrelia garinii--bactericidal activity of wild deer serum.

The susceptibility of Borrelia garinii to fresh wild deer sera was determined by incubating strain SIKA2 at 10% serum concentration for 1 hr at 37 C in an in vitro bactericidal assay. Each serum showed bactericidal effects at various levels. The effect was dependent on the concentration of antibody to the spirochetes. Complement was essential in the bactericidal assay because the inactivated deer serum showed greatly decreased activity. Our results suggest that B. garinii is sensitive to deer serum, in the presence of antibody and the bactericidal effect is important for preventing Lyme disease in wild sika deer.

Animals↗

Fimbria-specific immune response in various inbred mice inoculated with Porphyromonas gingivalis 381.

We studied the genetic control of Porphyromonas gingivalis fimbriae response. Inbred mice with different H-2 haplotypes and/or different genetic backgrounds were inoculated with viable P. gingivalis 381 cells and tested for fimbria-specific T cell responses in vivo (delayed-type hypersensitivity). H-2d mice showed a strong footpad response, whereas H-2b mice showed a weak response to fimbriae from P. gingivalis. Similar evidence of genetic control was obtained with an enzyme-linked immunosorbent assay for the detection of IgG antibody in inbred mice (BALB/c, C3H/HeN and C57BL/6). Several immunoglobulin G (IgG) subclass responses were associated with H-2 in these B10 congenic mice. However, quantification of IgG antibody to fimbriae was not controlled by H-2 in B10 congenic mice. The results indicate that, in mice, the responsiveness to fimbriae of P. gingivalis can be controlled by several genes, including the H-2 complex. C3H/HeN mice were inoculated with the fimbriae intravenously, and the expression of surface antigens on spleen T cells was measured in a fluorescent antibody cell sorter. Stimulation by fimbriae resulted in a changed expression of surface antigens on T cells. Thus, the fimbriae can induce T cell activation.

Animals↗

Experimental Borrelia garinii infection of Japanese quail.

Japanese quail inoculated subcutaneously with Borrelia garinii responded to infection. B. garinii was reisolated mainly from skin and randomly from several organs between 7 and 56 days postinoculation. Skin lesions were occasionally observed in association with spirochete recovery. All birds were positive for antibody at 53 to 56 days postinoculation. These results suggest that the Japanese quail is a model for experimental infection with B. garinii and that birds are reservoirs for Lyme disease spirochetes.

Animals↗

Both antisense and sense RNAs against the nucleocapsid protein gene inhibit the multiplication of mouse hepatitis virus.

DBT cells and several transfected cell lines which express antisense or sense RNA against the nucleocapsid protein gene of mouse hepatitis virus (MHV) were examined for the yields of MHV. The transfected cells showed 95 and 99% reduction of virus yield at 9 and 12 hr postinfection (p.i.) as compared with untransfected DBT cells. A remarkable decrease in MHV-specific RNA synthesis was observed in both transfected cell lines at 3.5 hr p.i. The result suggested that both antisense and sense RNAs inhibited viral replication at the initial stage of infection.

Animals↗

Subgingival temperature in rats with natural gingivitis.

Elevated temperature, normally a characteristic of inflammation, is a potential indicator of periodontal disease. The present investigation examined the relationship of subgingival temperature and various indicators of gingivitis in rats with spontaneous gingivitis (SUS rat) and in control rats (RES rat). In 24 SUS rats with gingivitis, the subgingival temperature rose, but not in 6 SUS and 20 RES rats with healthy gingiva. Subjects with high mean subgingival temperatures had significantly higher mean percentages of Prevotella intermedia and Fusobacterium nucleatum than age-matched RES rats. The results show that dental plaque, including specific bacteria P. intermedia and F. nucleatum, can play some role in causing the elevated temperatures.

Age Factors↗

Lyme disease spirochetes in a wild fox (Vulpes vulpes schrencki) and in ticks.

Lyme disease spirochetes were demonstrated in a wild female fox (Vulpes vulpes schrencki) and in Ixodes persulcatus ticks collected from the fox on Sapporo, Hokkaido, Japan. Spirochetes were detected in I. persulcatus, as well as skin lesions, brain, heart, kidney, and liver of the fox. Five of seven isolates reacted with a monoclonal antibody against Borrelia afzelii specific Osp B. Deoxyribonucleic acid (DNA) relatedness of a brain isolate was 89% to B. afzelii, and ranged from 50 to 67% to three other species. Immunoglobulin G antibodies to B. afzelii, B. garinii and B. burgdorferi sensulato, when tested in an enzyme-linked immunosorbent assay (ELISA), were negative in the fox. There were no antibodies against seven serovars of Leptospira interrogans.

Animals↗

Neutrophil dysfunction in rats with natural gingivitis.

The functions of polymorphonuclear neutrophils (PMN) from SUS rats with naturally occurring gingivitis were examined by the luminol-dependent chemiluminescence (CL), adherence and bactericidal tests. SUS rats with pre-gingivitis showed lower CL responses of isolated PMNs and whole blood than control rats (RES rats). After plague formation and progression of gingivitis, the CL response gradually increased in SUS rats. RES rats had healthy gingiva and showed no increase in CL responses. Impaired PMN adherence was observed in SUS rats with pre-gingivitis but not in RES rats. PMNs from SUS rats with pre-gingivitis also showed lower bactericidal activity than those from RES rats. Dysfunction of PMNs might induce gingivitis as a result of decreased protection against periodontal pathogens and an elevated level of CL response can be recognized with progression of gingivitis.

Animals↗

Demonstration of antigen-specific immune response against Streptococcus sanguis.

The genetic control of Streptococcus sanguis antigen response was studied. Mice sensitized with inactivated S. sanguis organisms antigen-injected at the base of the tail developed footpad swelling. Those with an I-Ak,q,r region of H-2 showed a strong footpad response, whereas those with an I-Ab,d,s region showed a weak response to S. sanguis cell wall antigen. Footpad response was mediated by CD4+,8- T cells by using in vitro monoclonal antibody treatment. Similar evidence of genetic control was obtained with an in vitro T cell proliferation assay. However, quantitation of antibodies against S. sanguis showed that antibody production was not controlled by H-2. These results indicated that both in vivo footpad swelling and in vitro T cell proliferation responses were functions of helper (CD3+,4+,8-) T cells and controlled by the I-A region of H-2.

Animals↗

Neurologic abnormalities in two dogs suspected Lyme disease.

A 2-year-old mongrel dog developed neurological signs following tick bite. These included astasia, persistent tonic convulsions and hyper-reflexia. Both serum IgG and IgM antibody titers against Borrelia burgdorferi were positive in enzyme-linked immunosorbent assay (ELISA). The neurological signs subsided after high-dose penicillin and streptomycin treatment. A strain of spirochetes (P427a) was isolated from the midgut of Ixodes persulcatus feeding on the dog. Morphological characteristic, immunological property and protein profile revealed that the isolate was B. burgdorferi. Similarly, a 2-year-old Labrador retriever dog developed neurological signs after tick bite and showed a positive IgG antibody titer against B. burgdorferi. Antibiotic treatment was effective also in this case. These findings suggest that neurological symptoms shown in both dogs were caused by infection with B. burgdorferi.

Animals↗

Similarity in nucleotide sequence of the gene encoding nontoxic component of botulinum toxin produced by toxigenic Clostridium butyricum strain BL6340 and Clostridium botulinum type E strain Mashike.

The complete nucleotide and deduced amino acid sequence of the nontoxic component of botulinum type E progenitor toxin is determined in recombinant plasmid pU9BUH containing about 6.0 kb HindIII fragment obtained from chromosomal DNA of Clostridium butyricum strain BL6340. The open reading frame (ORF) of this nontoxic component gene is composed of 3,486 nucleotide bases (1,162 amino acid residues). The molecular weight calculated from deduced amino acid residues is estimated 13,6810.1. The present study revealed that 33 nucleotide bases of 3,486 are different in the nontoxic component gene between C. butyricum strain BL6340 and C. botulinum type E strain Mashike. This corresponds to the difference of 17 amino acid residues in these nontoxic component.

Amino Acid Sequence↗

Serological survey for Borrelia burgdorferi infection in cattle in southern Hokkaido.

By means of the enzyme-linked immunosorbent assay (ELISA), antibodies to Borrelia burgdorferi were examined in 970 cattle from southern Hokkaido, Japan. The seropositivity rate in the cattle less than 2 years old differed significantly from that in those 3 years old or more. A seasonal variation was also observed in the rate of seropositivity to B. burgdorferi. High levels of anti-B. burgdorferi antibody were detected even in sera from a healthy group of cattle, indicating that there are likely to be many inapparent cases of B. burgdorferi infection. This article discusses the syndromes associated with Lyme borreliosis and addresses the issues of diagnosis and treatment in animals.

Animals↗

[Experimental animal models in Mongolian gerbils and mice].

Helicobacter pylori is an emerging candidate agent for the genesis of chronic gastritis and peptic ulcer. There have been many challenges to establish an experimental infection with H. pylori in animals. In recent, several studies reported for animal infection with this organism. One of them was the study using urinary system. The study indicated the ability of H. pylori to induce inflammation without significant low pH condition. Experimental infection in Mongolian gerbils showed that H. pylori could colonize on the surface of the gastric mucosa in animals without human. The study of nude mice by Karita et al. showed several valuable facts concerning the relation between bacterial pathogenicity and potential of bacterial colonization in stomach. The study in which remaining of bacteria was extended showed that colonization of H. pylori was affected by bacterial flora in the stomach.

Animals↗

Three types of binding by Porphyromonas gingivalis and oral bacteria to fibronectin, buccal epithelial cells and erythrocytes.

This study showed that the interaction of oral bacteria with fibronectin differed with the type of organism examined. Significant binding of fibronectin was found with Porphyromonas gingivalis non-fimbriated (F-) strain in comparison with the fimbriated strain (F+). However, the F+ strain adhered to buccal epithelial cells in significantly larger numbers than the F- strain. Fibronectin binding and epithelial cell adherence were not associated with haemagglutinating activity. These assays clearly define at least three distinct types of binding by oral bacteria: to fibronectin, buccal epithelial cells and erythrocytes.

Adhesins, Bacterial↗

Oligo-2',5'-adenylate synthetase activity in cells persistently infected with human T-lymphotropic virus type I (HTLV-I).

Spontaneous production of interferon-gamma (IFN-gamma) was shown in several T-lymphoblastoid cell lines persistently infected with human T-lymphotropic virus (HTLV-1). However, the produced IFN-gamma was not always associated with the induction of the antivirus state. The induction of oligo-2',5'-adenylate synthetase (2-5AS) by IFN was studied in five human T-cell lines persistently infected with HTLV-I (MT-1, MT-2, SMT-1, HUT 102 and OKM-2). Four cell lines are able to produce IFN-gamma spontaneously, while the OKM-2 cell line is not. Poor induction of 2-5AS was recognized in three (MT-1, MT-2 and SMT-1) of the four cell lines producing IFN-gamma, though the poor induction was improved after long-term cultivation of cells with IFN-alpha. On the contrary, in the OKM-2 cell line, significant activity of the enzyme was induced by IFN-alpha. Induction of 2-5AS was not correlated with cell growth inhibition, but with the antivirus state. Furthermore, an inverse relationship between IFN-gamma production and 2-5AS induction was demonstrated in these cell lines with the exception of HUT 102 cells.

2',5'-Oligoadenylate Synthetase↗

Seroepidemiological survey for antibody to Borrelia burgdorferi in cows.

Antibody to Borrelia burgdorferi was examined in 380 healthy and 38 clinical cases of cows from Hokkaido and Shizuoka in Japan. In healthy animals, IgG and IgM antibody to B. burgdorferi HO14 strain were found in 44 cows (14.6%) and 24 cows (8.0%) from Hokkaido. In contrast, antibody-positive case was not observed except for only 1 case which was IgM positive (1/79: 1.3%) in cows from Shizuoka. Mean antibody levels of healthy animals in Hokkaido and Shizuoka were 0.651 and 0.263 (IgG antibody to HO14 strain), 0.642 and 0.169 (IgG to HP3 strain), 0.613 and 0.367 (IgM to HO14 strain) and 0.582 and 0.286 (IgM to HP3 strain). The differences of the antibody levels between cows from Hokkaido and Shizuoka were significant. Seasonal difference was found in seropositive cows from Hokkaido. The rate of seropositive cows was high in summer (23.4% in June and 11.8% in July) but low in winter (0% in January and February). The pattern was discussed to be associated with activation of ticks. One of 4 cows with arthritis showed significantly higher IgG antibody level than that of healthy cows and cows with some disease, although the serum was collected from Shizuoka where antibody-positive animals for B. burgdorferi were rare among healthy cows. This high IgG antibody may suggest that the arthritis of such cows was caused by infection with B. burgdorferi. Two of 7 cows with unclassified abortion showed positive antibody reaction in Hokkaido. These cases, however, may not be related to the B. burgdorferi infection because the positive rate was similar to those of healthy cows in the same season.

Animals↗

Inhibition of mouse hepatitis virus multiplication by an oligonucleotide complementary to the leader RNA.

An oligonucleotide complementary to a leader RNA of positive-stranded mouse hepatitis virus (MHV) was tested for the effect on the viral multiplication in mouse DBT cells. A 14-mer antisense oligonucleotide contained a sequence complementary to the conserved pentanucleotide sequence, UCUAA, of the leader RNA. A treatment of MHV-infected cells with the antisense oligonucleotide at concentrations from 5 to 25 microM had an inhibitory effect on the viral multiplication and reduced the synthesis of viral specific mRNA and proteins. No inhibitory effect was observed when the cells were treated with sense oligonucleotide and oligonucleotide which contained unrelated sequences at concentrations from 1 to 10 microM. These results showed that antisense oligonucleotide against the leader RNA reduced the multiplication of positive-stranded RNA virus, MHV.

Animals↗

Detection of antibodies against Borrelia burgdorferi in patients with uveitis.

We determined the antibody response against Borrelia burgdorferi strains isolated from Japanese Ixodes ovatus and Ixodes persulcatus ticks by enzyme-linked immunosorbent assay and indirect immunofluorescence assay of serum specimens from 127 patients with uveitis. We examined samples of serum from Japanese patients with unclassified uveitis, iridocyclitis caused by herpes zoster virus, Behçet's disease, Vogt-Koyanagi-Harada syndrome, sarcoidosis, or other conditions (sympathetic ophthalmia, Posner-Schlossman syndrome and acute anterior uveitis with ankylosing spondylitis). Serum from healthy individuals and patients with Lyme disease served as negative and positive control samples, respectively. Significantly higher antibody titers were demonstrated in patients with uveitis than in control subjects. Of 29 patients with unclassified uveitis, nine (31) had significantly increased antibody titers against B. burgdorferi strain H014 by ELISA testing. Five patients also showed higher IgG and IgM responses than in three control subjects with Lyme disease. All positive controls showed joint problems characteristic of rheumatoid arthritis. One of three patients had uveitis. The patients were diagnosed as having Lyme disease on the basis of their history and serologic tests. A positive antibody response was recognized in several patients with Behçet's disease, Vogt-Koyanagi-Harada syndrome, sarcoidosis, and other conditions (acute anterior uveitis with ankylosing spondylitis), but not in control subjects.

Antibodies, Bacterial↗