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Biomedical subjects

T Masuzawa

Publications and source records attributed to T Masuzawa.

At least 73 records · Page 4Linked to original sources
PubMed1997

Identification of rickettsiae isolated in Japan as Coxiella burnetii by 16S rRNA sequencing.

The 16S rRNA genes of Japanese Coxiella isolates obtained from various sources and geographical areas were directly sequenced by dideoxynucleotide chain termination methods in which Taq DNA polymerase was used. The levels of sequence similarity among Japanese, European, and American isolates were more than 99%, and the Japanese isolates were identified as Coxiella burnetii, C. burnetii strains isolated worldwide, including Japan, were found to be very similar.

Animals↗
PubMed1997

A case of Lyme disease diagnosed serologically.

A 57-year-old Japanese male developed erythema migrans, arthralgia, and neurological symptoms after mountain climbing in Nagano Prefecture. On ELISA, his serum reacted to Borrelia isolated from various sources. The patient's serum reacted with a 41 kilodalton protein which was reactive with monoclonal antibody H9724 specific to flagellin of Borrelia on Western blotting. He recovered after taking minocycline 200 mg daily for 2 weeks.

Antibodies, Viral↗
PubMed1997

Consensus sequence on the genes encoding the major outer surface proteins (OspA and OspB) of Borrelia garinii isolate.

Japanese Lyme borrelias classified as ribotype IV is predominant among isolates derived from clinical specimens, reservoir rodents and Ixodes persulcatus ticks, and has been characterized as Borrelia garinii. These B. garinii isolates have antigenic and genetic features apparently different from North American, European and other Asian isolates, especially in major outer surface proteins A (OspA) and B (OspB). In this study, we cloned and sequenced the genes encoding OspA and OspB from B. garinii strain FujiP2 (ribotype IV strain) isolated from I. persulcatus in Shizuoka, Japan. A sequence analysis revealed significant differences to the previously published sequences of ospA and ospB of B. burgdorferi sensu lato. The open reading frames of ospA and ospB consist of 822 and 888 nucleotides corresponding to the proteins of 273 and 295 amino acids, with molecular weights of 29,643 and 31,786 daltons, respectively. The most interesting finding is that the two osp genes share a consensus 282 bp sequence in their carboxy-terminal portions and that the ospB gene is flanked by a 282 bp-long direct repeat sequence. The deduced amino-acid (aa) sequences of OspA and OspB of strain FujiP2 showed 60.1% homology, and have overall similarities of 70.5%, 70.3% and 75.6% to OspAB proteins of B. burgdorferi sensu stricto strain B31, Borrelia afzelii strain ACA1 and Borrelia garinii strain Ip90, respectively.

Amino Acid Sequence↗
PubMed1997

Genetic diversity of Borrelia burgdorferi sensu lato isolated in far eastern Russia.

One-hundred and fifty-seven Borrelia isolated from adult ticks, Ixodes persulcatus, and wild rodents, Clethrionomys rufocanus and Apodemus peninsulae, in the far eastern part of Russia were characterized and identified by restriction fragment length polymorphism (RFLP) of the 5S-23S rRNA intergenic spacer. Some isolates showed unique RFLP patterns and were determined as Borrelia garinii on the basis of a sequence analysis of the intergenic spacer amplicon and reactivity with species-specific monoclonal antibodies (MAbs). 86.5 and 12.7% of the tick isolates, and 74.2 and 12.9% of the rodent isolates were determined as Borrelia garinii and Borrelia afzelii, respectively, but not Borrelia burgdorferi sensu stricto was detected. This finding is similar to the results obtained from Borrelia surveys of I. persulcatus and wild rodents in Hokkaido, Japan.

Animals↗
PubMed1997

Deletion in the genes encoding outer surface proteins OspA and OspB of Borrelia garinii isolated from patients in Japan.

We detected the expression of outer surface proteins OspA and OspB, and characterized the genes encoding the two Osps of eight Borrelia garinii isolates from patients in Japan. Six of the eight strains shared a common antigenic epitope in their OspA and/or OspB proteins to monoclonal antibody P3134 against OspB, and were identified to have a conserved carboxyl terminus on their ospA and ospB genes by Southern blot hybridization. One strain, JEM4, did not express OspB protein, which was due to lack of the ospB gene. Gene cloning and sequencing analysis revealed that it had only one osp open reading frame with 819 nucleotides, which was similar to the ospA gene. The deletion of the ospB gene could be explained by a homologous recombination based on the common C-terminal sequences on the ospAB operon.

Antigens, Bacterial↗
PubMed1997

Negative finding in cross-protective activity of Japanese Borrelia isolates against infection with three species of Lyme disease Borrelia in outbred mice.

Outer surface protein A (OspA) is the most promising candidate for a component vaccine against Lyme disease caused by Borrelia burgdorferi sensu lato. Active cross-protection using a whole-cell vaccine prepared from strains belonging various OspA serotypes observed in Japan and worldwide was examined. No cross-protection was obtained by heterologous OspA-serotype vaccines. Since OspA is a highly variable protein expressed by Borrelia, this suggests that immunologically different OspA serotypes need to be combined for the development of an effective vaccine in Japan.

Animals↗
PubMed1997

Long-term evaluation of a nonpulsatile mechanical circulatory support system.

Antithrombogenicity of a centrifugal pump (CP) developed in our institute is provided by a central balancing hole (BH) in the impeller. A current CP, the National Cardiovascular Center (NCVC)-2, was ameliorated to improve antithrombogenicity, whereby the BH diameter was widened to improve self washout flow velocity, and an edge of the thrust bearing was rounded off to minimize flow separation. Effects of these modifications were assessed in a long-term in vivo experiment. The antithrombogenicity, hemolytic property, and mechanical durability of the NCVC-2 were investigated in 3 goats. The NCVC-2 was installed paracorporeally between the left atrium and the aorta and driven as long as possible at rotating speeds of about 2,800 rpm. The NCVC-2 ran for 50, 200, and 367+ days. The mean bypass flow rates were 6.8, 5.0, and 5.3 L/min, respectively. Creatinine, blood urea nitrogen (BUN), glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) did not increase until one week before termination. Plasma free hemoglobin was kept to a level less than 15 mg/dl, except for the last week of the second case. These results indicate that the NCVC-2 has excellent antithrombogenicity, an acceptable hemolytic property and the necessary durability for prolonged use.

Alanine Transaminase↗
PubMed1997

Noninvasive pump flow estimation of a centrifugal blood pump.

A flow rate estimating method was investigated for a centrifugal blood pump developed in our institute. The estimated flow rate was determined by the power consumption, the rotating speed of the motor, and the hematocrit value. The power consumption and the rotating speed of the motor were measured with a wattmeter. The examinations were performed in a closed mock loop filled with goat blood with hematocrit values of 21.5%, 28%, 34%, and 42%. Measured values of blood viscosity were 2.47, 3.09, 3.71, and 5.07 mPa.s at a share rate of 37.5/s, respectively. A linear correlation between the power consumption and the pump flow rate was observed in all hematocrit values. But variations in hematocrit caused a difference in the flow rate up to 1.1 L/min at the same power consumption and rotating speed. Effects of blood viscosity on the flow estimation were corrected by the hematocrit value. The value of the coefficient of determination, R2, between the estimated flow rate and the measured flow rate was 0.988. These results may indicate that the flow estimating method calculated by the power consumption of the motor, the rotating speed, and the hematocrit value is useful in the clinical situation.

Animals↗
PubMed1997

PCR-restriction fragment length polymorphism analysis of the ospC gene for detection of mixed culture and for epidemiological typing of Borrelia burgdorferi sensu stricto.

Restriction fragment length polymorphism (RFLP) analysis of the outer surface protein C (ospC) gene amplicon was used for rapid screening for genetic variability within Borrelia burgdorferi sensu stricto species and for detection of multiple borreliae in culture. Primers for the ospC gene amplified a fragment of about 600 bp from Borrelia cultures. After cleavage of the amplified products by MboI and DraI, eight different RFLP types were found among 13 B. burgdorferi sensu stricto strains from various sources and geographical areas, and three RFLP types were found among 10 representative isolates from skin biopsy specimens taken from patients residing on the eastern end of Long Island, New York (B. W. Berger, R. C. Johnson, C. Kodner, and L. Coleman, J. Clin. Microbiol. 30:359-361, 1992). These results suggested that the DNA organization of B. burgdorferi sensu stricto is heterogeneous not only globally but also within a localized geographical area and that the ospC-based typing approach could differentiate the B. burgdorferi sensu stricto. From the results obtained using mixed cultures of two different RFLP types of B. burgdorferi sensu stricto, contamination of at least 0.5% of different types of Borrelia cells in culture could be detected. This method could detect a multiple-B. burgdorferi sensu stricto infection in the bladders of mice experimentally infected with two different RFLP type strains. The present study showed that RFLP analysis of ospC-PCR products is a reliable method for epidemiological typing of B. burgdorferi sensu stricto and could be used for rapid detection of mixed Borrelia culture and multiple B. burgdorferi sensu stricto infections in animals, ticks, and patients.

Antigens, Bacterial↗
PubMed1997

Glycosphingolipid-binding protein of Borrelia burgdorferi sensu lato.

The binding of Borrelia burgdorferi, the causative agent of Lyme disease, to glycosphingolipids present in various types of cells was examined. B. burgdorferi bound specifically to galactosylceramide (GalCer) and glucosylceramide (GlcCer) but not to other glycosphingolipids, as determined by a thin-layer chromatography (TLC) overlay assay. The binding specificity of B. burgdorferi to various glycosphingolipids suggested that the binding receptor in this species is ceramide monohexoside. The levels of binding of B. burgdorferi virulent strain 297 to GlcCer, sulfatide, lactosylceramide, and galactosylgloboside were 56.2, 1.6, 15.9, and 9.7%, respectively, relative to that to GalCer. Virulent low-passage strains of B. burgdorferi were serially subcultured in BSK II medium, and the resultant high-passage strains were not capable of infecting mice and did not induce footpad swelling. The levels of binding of the low-passage strains to GalCer on TLC plates and to CHO-K1 cells in vitro were threefold higher than those of high-passage strains. Binding was not affected by pretreatment of Borrelia with monospecific anti-outer surface protein C (OspC) antiserum. These results indicated that the binding of Borrelia to glycosphingolipid expressed on the cell surface plays an essential role in infection of mammalian hosts. However, OspC was not associated with binding. The necessity of the sugar and N-acyl moieties in GalCer for the binding of Borrelia was shown by a TLC overlay assay using chemically modified GalCer. Furthermore, three proteins, 67-kDa protein, 62-kDa Hsp60, and 41-kDa flagellin, were involved in binding of B. burgdorferi to GalCer, as shown by blotting assay using biotinylated GalCer as a probe.

Animals↗
PubMed1997

[Present status of Lyme borreliosis and characterization of Lyme disease Borrelia isolated in Japan].

Lyme disease is a multisystemic disorder caused by infection with Borrelia burgdorferi sensu lato which is carried by ticks of the Ixodes ricinus complex. The agent was discovered in 1982 in North America and the disease is recognized as an emerging infectious diseases in North America and Europe. Japanese Borrelia isolates were characterized by genetic and immunologic analysis. Isolates from Ixodes ovatus were found to be unique by DNA/DNA hybridization analysis, restriction fragment length polymorphism analyses of the flagellin gene and the 16S rRNA genes, and were described as new species, Borrelia japonica. Isolates from Ixodes persulcatus were determined as Borrelia garinii and Borrelia afzelii. However, B. garinii found in Japan was different from those from Europe in immunologic and genetic characteristics of outer surface protein A, but B. afzelii isolates from Japan and Europe were identical. An experimental model of arthritis related to Lyme disease using outbred ddY mice was established. Whole cell vaccine prepared from North American and European isolates could not elicit protective immunity against infection of Japanese isolates. This implies that vaccine development using Japanese isolates is necessary. Borrelia bound specifically to galactosylceramide (GalCer), glucosylceramide and lactosylceramide which are present in various types of cells as binding receptor, but not to other glycosphingolipids. Furthermore, the infectivity of Borrelia may be associated with the binding to glycosphingolipids on the cell surface and a 67 kilodalton protein of Lyme disease Borrelia may be involved in binding of Borrelia to GalCer.

Animals↗
PubMed1997

Intracerebral Schwannoma--case report.

A 15-year-old girl presented with a rare intracerebral schwannoma manifesting as epileptic seizure. Computed tomography, magnetic resonance imaging, and cerebral angiography showed a right parietooccipital lobe tumor. The preoperative diagnosis was malignant glioma, but immunohistochemical and ultrastructural examinations showed the tumor was indistinguishable from peripheral schwannoma. Only 29 cases of schwannomas not related to the cranial nerve have been reported, mostly in children and young adults. The origin appears to be ectopic or perivascular elements in the brain. Such intracerebral schwannomas can be classified according to origins into intra-axial, periventricular, dural attachment, and other types.

Adolescent↗
PubMed1996

Intracellular inositol 1,3,4,5-tetrakisphosphate enhances the calcium current in hippocampal CA1 neurones of the gerbil after ischaemia.

1. To examine the role of the phosphoinositide cascade triggered by disturbed Ca2+ homeostasis in ischaemic neurones, inositol 1,3,4,5-tetrakisphosphate (InsP4) was applied to the cytoplasmic face of membrane patches isolated from CA1 pyramidal neurones in the gerbil hippocampus. 2. In outside-out recordings, InsP4 induced an inward current which was increased by raising the extracellular [Ca2+]. In contrast, no clear channel openings could be observed in patches from neurones of sham-operated gerbils. 3. Open probabilities of InsP4-activated channels were significantly decreased upon application of omega-conotoxin but were not affected by omega-agatoxin or nifedipine. 4. In inside-out patches using high concentrations of Ca2+, Ba2+ or Sr2+ in the pipette solution, InsP4 enhanced inward currents. 5. Application of the isomers of InsP4 slightly enhanced the currents, but inositol 1,4,5-trisphosphate (InsP3) had no effect. 6. In the absence of InsP4 there was a single main Ba2+ current peak of 4.0 pA in amplitude, whereas upon its application two main peaks of 3.0 and 7.2 pA were present. 7. The open probabilities of these channels were apparently increased by InsP4. 8. These findings support the view that a disturbed phosphoinositide cascade occurs in the hippocampal pyramidal neurones after ischaemia and the InsP4 thus formed plays an important role in promoting the Ca2+ accumulation which results in neuronal death.

Animals↗
PubMed1996

Elevated expression of DNA polymerase beta gene in glioma cell lines with acquired resistance to 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3- nitrosourea.

We investigated the expression of DNA polymerase beta (beta-pol) and O6-methylguanine-DNA methyltransferase (MGMT) in human glioma cells with acquired resistance to 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosoure a (ACNU) and in the parent cells. ACNU-resistant T430 (T430R) and A172 (A172R) glioma cell lines were established following repeated exposure to ACNU. The level of MGMT mRNA expression was elevated in T430R, but not in A172R. In contrast, the level of beta-pol mRNA expression and the level of beta-pol protein were elevated in A172R, compared with the parent cells. While the mechanism of MGMT repair has been considered to be important in the drug resistance of human brain tumors to ACNU, our present results demonstrate that beta-pol may also play an important role in the acquisition of tumor cell resistance to ACNU in human gliomas.

Antineoplastic Agents↗
PubMed1996

Characterization of Borrelia sp. isolated from Ixodes tanuki, I. turdus, and I. columnae in Japan by restriction fragment length polymorphism of rrf (5S)-rrl (23S) intergenic spacer amplicons.

Borrelia isolated from various sources in Japan, including rare species of ixodid ticks, Ixodes tanuki, I. turdus, and I. columnae, were characterized by restriction fragment length polymorphism analysis and sequencing analysis of the 5S-23S rRNA intergenic spacer amplicon. Borrelia sp. isolated from I. tanuki, I. turdus and I. columnae generated restriction fragment length polymorphism patterns different from those of known B. burgdorferi sensu lato isolates previously reported. Furthermore, some B. afzelii and B. garinii isolated in Japan showed unique RFLP patterns which were not observed among European B. afzelii and B. garinii.

Animals↗
PubMed1996

Involvement of specific macrophage-lineage cells surrounding arterioles in barrier and scavenger function in brain cortex.

The transport of solutes between blood and brain is regulated by a specific barrier. Capillary endothelial cells of brain are known to mediate barrier function and facilitate transport. Here we report that specific cells surrounding arterioles, known as Mato's fluorescent granular perithelial (FGP) cells or perivascular microglial cells, contribute to the barrier function. Immunohistochemical and in situ hybridization studies indicate that, in normal brain cortex, type I and type II macrophage scavenger receptors are expressed only in FGP/perivascular microglial cells, and surface markers of macrophage lineage are also detected on them. These cells mediate the uptake of macromolecules, including modified low density lipoprotein, horseradish peroxidase, and ferritin injected either into the blood or into the cerebral ventricles. Accumulation of scavenged materials with aging or after the administration of a high-fat diet results in the formation of honeycomb-like foam cells and the narrowing of the lumen of arterioles in the brain cortex. These results indicate involvement of FGP/perivascular microglial cells in the barrier and scavenger functions in the central nervous system.

Adult↗
PubMed1996

Extensive RNA editing of U to C in addition to C to U substitution in the rbcL transcripts of hornwort chloroplasts and the origin of RNA editing in green plants.

We cloned and sequenced a portion of chloroplast DNA from the hornwort Anthoceros formosae. A nucleotide sequence of 7556 bp contained structures similar to those of ndhK, ndhC, trnV, trnM, atpE, atpB, rbcL, trnR and accD. The arrangement of these was the same as that of other chloroplast DNA. However, two nonsense codons were located within the putative coding region of rbcL, although they were used as putative termination codons of the genes. RNA was extensively edited in the transcripts of rbcL when cDNA sequences were analyzed. The unusual nonsense codons of TGA and TAA became CGA and CAA respectively. These are examples of U to C type RNA editing, which was never been found before in chloroplast mRNA. In general, 13 Cs of genomic DNA were found as Ts in the cDNA sequence and seven Ts were found as Cs. This is the first finding of RNA editing on the transcripts of rbcL and also in bryophytes. This event had been thought to arise in land plants after the split of bryophytes. The origin of RNA editing is discussed in relation to the landing of green plants.

Amino Acid Sequence↗