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Biomedical subjects

T Mikami

Publications and source records attributed to T Mikami.

At least 271 records · Page 15Linked to original sources
PubMed1997

Changes in cerebral blood flow velocity in healthy young men during overnight sleep and while awake.

Using transcranial Doppler ultrasonography, we measured in 6 healthy young men cerebral blood flow velocity (CBFV) in the middle cerebral artery as well as oxygen saturation by a non-invasive method. Continuous recordings were taken starting from a point before the onset of sleep, throughout the duration of normal nighttime sleep, ending after awakening. During stages 2, 3 and 4, CBFV was approximately 15% lower than during the waking period preceding sleep. CBFV during rapid eye movement sleep did not differ from the presleep waking value, whereas the postsleep waking value was 6.6% lower. In 5 subjects CBFV showed a transient rise after sleep onset. Oxygen saturation was lower during sleep than during waking.

Adult↗
PubMed1997

Galactosamine-induced apoptosis in the primary mouse hepatocyte cultures.

Galactosamine (GalN)-induced apoptosis was investigated in cultured hepatocytes from mice. The percentage of fragmented DNA measured by the ELISA method increased in a concentration-dependent manner from the very early stage, i.e. 0.5 hrs, after GalN-exposure. In addition, a ladder-like fragmentation pattern by agarose gel electrophoresis appeared first at 3 hr-exposure to 20 mM GalN, at 6 hr-exposure to 10 mM GalN and at 12 hr-exposure to 5 mM GalN, respectively. On the other hand, cytotoxicity indicated by leakage of lactate dehydrogenase from cultured hepatocytes to culture medium was first detected at 24 hrs after GalN-treatment. Morphologically, formation of blebs and apoptotic bodies was observed from 12 hr-exposure to 20 mM GalN and from 24 hr-exposure to 10 mM GalN, respectively. Thus GalN could induce apoptosis in primary hepatocyte cultures from mice.

Animals↗
PubMed1997

Identification and DNA sequence analysis of the Marek's disease virus serotype 2 genes homologous to the thymidine kinase and UL24 genes of herpes simplex virus type 1.

The thymidine kinase (TK) gene has been used as a safe and convenient locus for expression of heterologous proteins in some alphaherpesviruses including herpesvirus of turkeys (HVT) antigenically related to Marek's disease virus (MDV) serotypes 1 (MDV1) and 2 (MDV2). In MDV2 strain HPRS 24 genome, genes equivalent to the TK and UL24 homologues of herpes simplex virus type 1 were identified and sequenced. The MDV2 UL24 gene overlaps the 5' end of the TK gene in a head-to-head orientation. The predicted region encoding for the MDV2 TK gene is 1,056 nucleotides, corresponding to a polypeptide of 352 amino acids in length. Putative nucleotide- and thymidine-binding sites were identified within the predicted amino acid sequence. The predicted region encoding for the UL24 gene is 948 nucleotides, corresponding to a polypeptide of 316 amino acids in length. By northern blot analyses using MDV2 TK- and UL24-specific DNA probes, four transcripts of approximately 7.8, 5.0, 3.5, and 1.1 kb for the TK gene, and a transcript of 3.8 kb for the UL24 gene were detected in MDV2-infected cells. Alignment of the amino acid sequence of MDV2 TK homologue with those published for TK homologues of other MDV serotypes showed 73.9% (MDV1 vs. MDV2), 58.2% (MDV1 vs. HVT), and 56.8% (MDV2 vs. HVT) identities. Comparison to other alphaherpesvirus TK homologues revealed amino acid sequence homologies varying from 34.5% to 27.8%. The putative MDV2 UL24 homologous protein had identity with the well conserved five motifs among alphaherpesviruses.

Amino Acid Sequence↗
PubMed1997

Identification and characterization of the feline herpesvirus type 1 glycoprotein C gene.

The feline herpesvirus type 1 (FHV-1) gene encoding glycoprotein C (gC) has been sequenced and identified based on its genomic location and comparative analysis to other alphaherpesvirus gCs, and the expressed gC protein was also identified by using specific monoclonal antibodies. The FHV-1 gC gene was located within a 7.0 kbp EcoRI fragment, and was 1602 bp in length. The amino acid sequence deduced from the nucleotide sequence was predicted to encode a membrane glycoprotein containing a characteristic N-terminal hydrophobic signal sequence, nine potential N-linked glycosylation sites, and C-terminal transmembrane and cytoplasmic domains. The FHV-1 gC was expressed in COS-7 cells. When flowcytometric analysis was carried out, the gC expressed in COS-7 cells reacted with a panel of monoclonal antibodies against gp113: By immunoprecipitation analysis, the gC expressed in COS-7 cells possessed molecular masses of 125-150 kilodalton, and was similar in size to that in FHV-1-infected CRFK cells.

Alphaherpesvirinae↗
PubMed1997

High incidence of positive autoantibodies against thyroid peroxidase and thyroglobulin in patients with sarcoidosis.

OBJECTIVE: Although abnormalities of the humoral immune system, such as increased immunoglobulin production, are known in sarcoidosis, the relationship between sarcoidosis and autoimmune disorders in uncertain. We studied the incidence of thyroid autoantibodies and the prevalence of Hashimoto's thyroiditis in patients with sarcoidosis. PATIENTS AND MEASUREMENTS: Sixty-two patients with pulmonary sarcoidosis, diagnosed by a combination of clinical, radiographic and histological findings were studied. As controls, three groups of subjects aged 40 and over without a known history of thyroid disease (60 patients with pulmonary diseases other than sarcoidosis, 88 hospital employees and 82 company workers), were also analysed. Antibodies against thyroid peroxidase (TPO-Ab) and purified thyroglobulin (Tg-Ab) were measured by radioimmunoassay and antibodies against microsomal antigen (MCHA) and thyroglobulin (TGHA), by haemagglutination. RESULTS: Seventeen of 62 patients (27.4%) had either positive TPO-Ab or Tg-Ab or both. All the patients with positive thyroid autoantibodies were of middle or advanced age, and the incidence of positive TPO-Ab/Tg-Ab in patients with sarcoidosis aged 40 and over was 54.5% in males, 32.4% in females and 37.8% overall. The prevalence was significantly higher in males compared to age-matched control males (0-7.7% in the controls), and in female patients was twice that found in controls (11.8-16.3%). Seven patients had Hashimoto's thyroiditis, indicating that the prevalence was 11-3%, and much higher than that previously reported. CONCLUSIONS: The data show a remarkably high incidence of thyroid autoantibodies in patients of middle of advanced age with sarcoidosis, especially in males, and a higher prevalence of Hashimoto's thyroiditis than in previous reports.

Age Factors↗
PubMed1997

Occurrence of nonenzymatic N-acetylation of sphinganine with acetyl coenzyme A producing C2-H2-ceramide and its inconvertibility to apoptotic C2-ceramide.

Sphinganine, a biosynthetic precursor of ceramide, was non-enzymatically acetylated with acetyl coenzyme A at the C-2-amino residue to produce C2-H2-ceramide (N-acetyl sphinganine) in an organic solvent and in an aqueous solution with a high yield, whereas sphingenine was only acetylated slightly. The structure of the N-acetyl sphinganine was identified with mass spectrum, and with chromatography using an authentic N-acetylated substance. Furthermore, the C2-H2-ceramide was examined for enzymatic desaturation to determine whether C2-ceramide, a cell-permeable ceramide responsible for apoptosis of cells, was produced, revealing an inferior substrate for H2-ceramide desaturase of horse brain microsomes.

Acetyl Coenzyme A↗
PubMed1997

Characterization of several pseudorabies viral strains by virus-neutralization test using monoclonal antibodies.

In the present study, five mouse monoclonal antibodies (MAbs) to the pseudorabies virus (PRV) Yamagata-81 strain were produced. The MAbs were used in cross-neutralization tests and cross-indirect enzyme-linked immunosorbent assay (ELISA) against three PRV viral strains isolated in Argentina and another four obtained from the United States, Japan, France, and Sweden. Four of five MAbs needed the presence of complement to produce or enhance neutralization activity. No differences were observed by ELISA. The MAbs showed different neutralizing activity against PRV strains, suggesting phenotypic heterogeneity among them.

Animals↗
PubMed1997

Gastrointestinal autonomic nerve tumor with giant abscess. A case report and literature review.

We report a gastrointestinal autonomic nerve tumor of the stomach with a giant abscess. The patient had fever and pain and was found to have anemia and an abdominal mass. X-ray and endoscopic examination showed a gastric submucosal tumor with a fistula to the gastric lumen. Partial gastrectomy was performed and no metastasis was found. On gross examination, the excised tumor was seen to be a submucosal solid tumor with a giant abscess. Alpha streptococci and anaerobic gram-negative rods were cultured from the pus of the abscess. The tumor resembled a gastric myogenic tumor composed of spindle cells, partly showing storiform and epithelioids. Tumor cells showed positive staining for vimentin and neuron-specific enolase but were negative for desmin, alpha-smooth muscle actin, and S-100 protein. Ultrastructural examination showed remarkable interdigitation of cytoplasmic processes with neurosecretory granules between the tumor cells. This lesion was similar to previously described gastrointestinal autonomic nerve tumors. Gastrointestinal autonomic nerve tumors are a rare, distinct subtype of gastrointestinal stromal tumors; although several cases of focally necrotic tumors have been reported, there has been only one report of the tumor with an abscess, as in our case.

Abscess↗
PubMed1997

Mapping of antigenic sites involved in neutralization on the capsid protein of feline calicivirus.

In order to locate amino acid residues involved in the formation of feline calicivirus (FCV) neutralizing epitopes, we analysed the capsid protein gene of monoclonal antibody neutralization-resistant variants of FCV. Amino acid substitutions in the variants were identified in the two hypervariable regions of the capsid protein. Four linear and two conformational epitopes were located in the regions from residues 426 to 460 and 490 to 520, respectively. The relative positions of individual epitopes agreed with our previous antigenic analysis. Two antigenic sites composed of the neutralizing epitopes were mapped in the hypervariable regions of the capsid protein, demonstrating that a relationship exists between the genetic variability and antigenic differences in the neutralization of FCV.

Animals↗
PubMed1997

Molecular and phylogenetic analyses of the haemagglutinin (H) proteins of field isolates of canine distemper virus from naturally infected dogs.

We isolated three strains of canine distemper virus (CDV)--the Ueno, Hamamatsu, and Yanaka strains--from dogs in Japan and analysed the molecular properties of their haemagglutinin (H) proteins. Immunoprecipitation of all three strains with a monoclonal antibody revealed H proteins with molecular masses of 84 kDa, which differs from the molecular mass (78 kDa) of the H protein of the Onderstepoort vaccine strain. However, after tunicamycin treatment immunoprecipitation identified H proteins of identical molecular mass (68 kDa) for all three field isolates and the vaccine strain. Sequence analysis showed nine potential sites for asparagine-linked glycosylation in the H proteins of the new isolates, in contrast to four in the H protein of the Onderstepoort strain. Thus, variation in glycosylation of the H proteins of the isolates and the vaccine strain may cause differences in antigenicity of the viruses. Sequences of the H genes showed that the new Japanese isolates have 99% identity with each other, 95% with other European and American isolates (from seals, a German dog, a ferret and large felids) and 90% with the vaccine strain. Phylogenetically, the new Japanese isolates form one cluster which is separate from recent European or American isolates, all of which are distinct from vaccine strains.

Amino Acid Sequence↗
PubMed1997

Replication of feline syncytial virus in feline T-lymphoblastoid cells and induction of apoptosis in the cells.

Feline syncytial virus (FSV) was isolated from feline peripheral blood mononuclear cells of FSV-seropositive cats. When the susceptibility of feline T-lymphocytes to FSV was examined using three strains of FSV, FSV antigens were detected in the FSV-infected T-lymphoblastoid cells. Further, a diversity of biological properties, including replication kinetics and syncytia formation, was noted among the strains, and condensation of chromatin and the fragmentation of cellular DNA were observed in the infected cells. From these data, we conclude that FSV is lymphotropic and can induce apoptosis in the lymphocytes.

Animals↗
PubMed1997

Antitumor activity of TZT-1027, a novel dolastatin 10 derivative.

Dolastatin 10, a pentapeptide isolated from the marine mollusk Dolabella auricularia, has antitumor activity. TZT-1027, a dolastatin 10 derivative, is a newly synthesized antitumor compound. We evaluated its antitumor activity against a variety of transplantable tumors in mice. Intermittent injections of TZT-1027 were more effective than single or repeated injections in mice with P388 leukemia and B16 melanoma. Consequently, TZT-1027 shows schedule dependency. TZT-1027 was effective against P388 leukemia not only when administered i.p., but also when given i.v. However, although TZT-1027 given i.v. was active against murine solid tumors, TZT-1027 administered i.p. was ineffective against all the tumors tested with the exception of colon 26 adenocarcinoma. The i.v. injection of TZT-1027 at a dose of 2.0 mg/kg remarkably inhibited the growth of three murine solid tumors; colon 26 adenocarcinoma, B16 melanoma and M5076 sarcoma, with T/C values of less than 6%. The antitumor activities of TZT-1027 against these tumors were superior or comparable to those of the reference agents; dolastatin 10, cisplatin, vincristine, 5-fluorouracil (5-FU) and E7010. In experiments with drug-resistant P388 leukemia, TZT-1027 showed good activity against cisplatin-resistant P388 and moderate activity against vincristine- and 5-fluorouracil-resistant P388, but no activity against adriamycin-resistant P388. TZT-1027 was also effective against human xenografts, that is, tumor regression was observed in mice bearing MX-1 breast and LX-1 lung carcinomas. TZT-1027 at 10 microM almost completely inhibited the assembly of porcine brain microtubules. Therefore, its mechanism of antitumor action seems to be, at least in part, ascribable to the inhibition of microtubule assembly. Because of its good preclinical activity, TZT-1027 has been entered into phase I clinical trials.

Animals↗
PubMed1997

Possible role of macrophage-derived soluble mediators in the pathogenesis of encephalomyocarditis virus-induced diabetes in mice.

Pancreatic islets from DBA/2 mice infected with the D variant of encephalomyocarditis (EMC-D) virus revealed lymphocytic infiltration with moderate to severe destruction of pancreatic beta cells. Our previous studies showed that the major population of infiltrating cells at the early stages of infection is macrophages. The inactivation of macrophages prior to viral infection resulted in the prevention of diabetes, whereas activation of macrophages prior to viral infection resulted in the enhancement of beta-cell destruction. This investigation was initiated to determine whether macrophage-produced soluble mediators play a role in the destruction of pancreatic beta cells in mice infected with a low dose of EMC-D virus. When we examined the expression of the soluble mediators interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in the pancreatic islets, we found that these mediators were clearly expressed at an early stage of insulitis and that this expression was evident until the development of diabetes. We confirmed the expression of these mediators by in situ hybridization with digoxigenin-labelled RNA probes or immunohistochemistry in the pancreatic islets. Mice treated with antibody against IL-1beta or TNF-alpha or with the iNOS inhibitor aminoguanidine exhibited a significant decrease in the incidence of diabetes. Mice treated with a combination of anti-IL-1beta antibody, anti-TNF-alpha antibody, and aminoguanidine exhibited a greater decrease in the incidence of disease than did mice treated with one of the antibodies or aminoguanidine. On the basis of these observations, we conclude that macrophage-produced soluble mediators play an important role in the destruction of pancreatic beta cells, resulting in the development of diabetes in mice infected with a low dose of EMC-D virus.

Animals↗
PubMed1997

Pulmonary alveolar proteinosis: diagnosis using routinely processed smears of bronchoalveolar lavage fluid.

AIMS: For the diagnosis of pulmonary alveolar proteinosis from bronchoalveolar lavage specimens it is normally necessary to make an ultrastructural examination. However, this is thought to be impractical for bronchoalveolar lavage specimens that have been routinely fixed in ethanol. In the present study, bronchoalveolar lavage cytology smears on slide glasses were examined directly ultrastructurally to make a diagnosis of pulmonary alveolar proteinosis. METHODS: Bronchoalveolar lavage smears from three pulmonary alveolar proteinosis patients were stained with Papanicolaou and periodic acid-Schiff (PAS) for identification of amorphous globular structures. Subsequently, they were refixed with glutaraldehyde and osmium tetroxide, and embedded in epoxy resin. Ultrathin sections were cut and examined ultrastructurally. RESULTS: Papanicolaou stained specimens from pulmonary alveolar proteinosis patients contained scattered amorphous or granular globules, 20-50 microns in diameter, which were PAS positive. Ultrastructural examination of the globules revealed multilamellated structures, characteristic of pulmonary alveolar proteinosis, in all cases. CONCLUSIONS: In general, it is thought that the morphological diagnosis of pulmonary alveolar proteinosis from bronchoalveolar lavage specimens requires both cytological and ultrastructural examination. However, the amorphous globules evident on cytology smears proved to contain multilamellated structures so that they can themselves be used as diagnostic evidence.

Adult↗
PubMed1997

Anti Candida activity of induced transferrin in mice immunized with inactivated Candida albicans.

Mice immunized with formalin-killed Candida albicans were resistant to challenge by a lethal amount of viable C. albicans. The growth-inhibitory activity to C. albicans was detected in sera from the immunized mice, and was inhibited by the addition of anti-transferrin antibody or ferric sulfate. Both the amount of transferrin and the unsaturated iron-binding capacity (UIBC) in the serum were significantly increased, indicating that apo-transferrin increased in the immunized mice. Moreover, the intraperitoneal administration of apo-transferrin enhanced the protection from the Candida infection in vivo.

Animals↗
PubMed1997

Characterization of canine herpesvirus glycoprotein D (hemagglutinin).

Glycoprotein D (gD) of canine herpesvirus (CHV) YP2 strain was expressed in COS-7 and insect (Spodoptera frugiperda; Sf9) cells. The gDs expressed in COS-7 and Sf9 cells reacted with a panel of monoclonal antibodies (MAbs) against CHV gD (hemagglutinin) and an MAb 25C9 against feline herpesvirus type 1 (FHV-1) gD by indirect immunofluorescence assay, and possessed a molecular weight (MW) of approximately 51-55 and 41-46 kilodalton (kDa), respectively, when examined by immunoblot analysis. After treatment with tunicamycin, the MW of the gD expressed in Sf9 cells became approximately 37 kDa. By hemadsorption (HAD) tests using canine or feline red blood cells (RBC), COS-7 cells expressing CHV gD adsorbed only canine RBC, but not feline RBC, whereas control COS-7 cells expressing FHV-1 gD adsorbed feline RBC, but not canine RBC. By hemagglutination (HA) tests, lysates of Sf9 cells expressing CHV gD agglutinated canine RBC, but not feline RBC. These HA and HAD activities were inhibited by HA-inhibition MAbs against CHV gD. Control lysates of Sf9 cells expressing FHV-1 gD agglutinated only feline RBC. Serum from mice inoculated with lysates of Sf9 cells expressing CHV gD possessed a high titer of virus-neutralizing activities against CHV infection. These results indicated that CHV gD is structurally similar to FHV-1 gD, but is functionally different from FHV-1 gD.

Alphaherpesvirinae↗
PubMed1997

Role of one N-linked oligosaccharide chain on canine herpesvirus gD in its biological activity.

The YP11mu strain of a plaque-selected canine herpesvirus (CHV) encoded a smaller molecular weight (MW) of gD than those of other strains including YP2 strain (Xuan et al., 1990). When nucleotide sequence of the mutated gD of YP11mu strain (gD(YP11mu)) was compared with that of gDs of other CHV strains, gD(YP11mu) lacked 12 nucleotides encoding 4 amino acids, NKTI, including one predicted potential N-linked glycosylation site and no other change was found in other regions. When the gD(YP11mu) and gD of YP2 strain (gD(YP2)) expressed in COS-7 and insect (Spodoptera frugiperda; Sf9) cells were compared each other, both gDs reacted with a panel of monoclonal antibodies (MAbs) against CHV gD by indirect immunofluorescence analysis and the gD(YP11mu) possessed an MW of approximately 47-51 and 39-44 kDa in COS-7 and Sf9 cells, respectively, which were smaller than the expressed gD(YP2) (approximately 51-55 and 41-46 kDa, respectively) by immunoblot analysis. After treatment with tunicamycin, the MW of both gDs in Sf9 cells became approximately 37 kDa. When hemagglutination (HA) test using canine red blood cells (RBC) were carried out, lysates of Sf9 cells expressing CHV gDs agglutinated canine RBC. Serum from mice inoculated with lysates of Sf9 cells expressing the gDs possessed a high titer of virus-neutralizing (VN) activities against CHV. These results indicated that the deletion of 4 amino acids possessing approximately 4 kDa of glyco-chain from gD of CHV in mammalian cells does not affect HA activity and VN antibody-inducing activity and that this deletion of gD(YP11mu) might be a good selective marker for development of recombinant viruses as a live vaccine.

Amino Acid Sequence↗