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Biomedical subjects

H Miyazawa

Publications and source records attributed to H Miyazawa.

At least 109 records · Page 6Linked to original sources

Measurement of serum IgE antibodies against Japanese cedar pollen (Cryptomeria japonica) in Japanese monkeys (Macaca fuscata) with pollinosis.

IgE antibodies against allergens of Japanese cedar (Cryptomeria japonica, CJ) pollen in the serum of seven Japanese monkeys (Macaca fuscata) with pollinosis were measured by fluorometric indirect enzyme-linked immunosorbent assay (ELISA). All of the monkeys were found to have specific IgE to the crude pollen antigen. The specific IgE levels were well correlated with those determined by the Pharmacia CAP system. IgE antibodies were then assayed with two kinds of purified allergens (Cry j I and Cry j II) by the ELISA. We found that five monkeys had specific IgE to both allergens, although the other two had IgE only to Cry j I or Cry j II; there is different immune responsiveness to the two major allergens in the monkeys.

Allergens↗

[Detection of mediastinal lymph node metastasis from lung cancer with positron emission tomography (PET) using 11C-methionine].

Twenty-five patients with primary non-small cell lung cancer underwent the positron emission tomography (PET) using 11C-methionine to detect the mediastinal lymph node metastasis. We introduced the positron angiography to recognize precisely the anatomical orientation of the mediastinal lymph nodes. The 11C-uptake of the lymph node was expressed with distribution absorption ratio (DAR). A total 107 lymph nodes were examined. The average DAR in metastatic lymph nodes (n = 28) was 3.89 while that of non-metastatic nodes (n = 79) was 2.38 indicating a significant difference (p < 0.001). The most adequate threshold for detection of metastasis was 3.3 with sensitivity of 100%, and specificity of 87.3% and overall accuracy of 89.7%. Metastasis of squamous cell carcinoma was diagnosed more accurately than that of adenocarcinoma. Thus, PET using 11C-methionine may offer a new method to detect the mediastinal lymph node metastasis from lung cancer.

Adult↗

[The study of pulmonary tuberculosis in the elderly].

A clinical study of 52 patients diagnosed as having pulmonary tuberculosis at National Chiba-Higashi Hospital between 1988 and 1990 was performed. The cases ranged in age from 80-89 years; mean male patient age was 82.5, and mean female patient age was 84.3. Diagnosis of the cases were as follows: 19 discovered when checking into hospital because of chest symptoms; 14 diagnosed during the treatment of other diseases; 14 diagnosed during admission to the hospital for other diseases; and five cases were detected by chest X-rays. A total of 38 cases, had received primary treatment for pulmonary tuberculosis, 11 cases had received secondary treatment, and three cases were receiving treatment for tuberculosis. Upon admission to the hospital after the detection of tuberculosis, 19 cases tested positive to sputum smear examinations, six cases tested positive to culture examinations but negative to smear examinations, and culture examinations were negative in 27 cases. Regarding the chest X-ray findings, using the criterion of roentgenological classification for pulmonary tuberculosis established by the Japanese Society for Tuberculosis, two cases revealed type I, 29 cases revealed type II, and 21 cases revealed type III. Cavitary cases were observed in 60% of the chest X-ray findings. Upon hospital admission, 18 cases were observed to have circulatory diseases, 16 cases had central nervous diseases, 12 cases had digestive diseases and 11 cases had respiratory diseases. Nine cases had malignant neoplasm, five cases had diabetes mellitus and 14 cases had other diseases. A total of 18 cases ended in death; six cases died of pulmonary tuberculosis, and 12 cases died of other diseases.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

The c-myb proto-oncogene product binds to but does not activate the promoter of the DNA polymerase alpha gene.

The myb proto oncogene product (c-Myb) is a transcriptional regulator and its expression and function are tightly linked to the cellular entry into S phase and DNA synthesis. It has been shown [Venturelli, D., Travali, S. & Calabretta, B. (1990). Proc. Natl. Acad. Sci. USA, 87, 5963-5967] that inhibition of T-cell proliferation by a myb antisense oligomer is accompanied by down-regulation of DNA polymerase alpha expression. To examine whether the transcription of the DNA polymerase alpha gene is directly regulated by c-Myb, we have identified and characterized the 5' regulatory region of the human DNA polymerase alpha gene. Two major and several minor transcription start sites were identified by nuclease S1 mapping. DNA sequence analysis showed that the promoter region contains no TATA box, one CCAAT box and putative Ap-1, AP-2 and E2F binding sites. In DNAase I footprinting, the bacterially expressed c-Myb protected six sites in the 5' flanking region of the human DNA polymerase alpha gene. However, c-Myb did not activate the DNA polymerase alpha gene promoter in a co-transfection assay. Our results suggest that an unknown factor(s) is required for the c-Myb-induced activation of the DNA polymerase alpha gene promoter, or c-Myb does not directly activate this promoter.

Base Sequence↗

[Two cases of broncholithiasis removed by bronchofiberscopy].

Two cases of broncholithiasis, removed bronchoscopically, are reported. Case 1 was a 38-year-old female who was admitted with hemoptysis. The chest tomogram showed calcification near the right middle lobe bronchus. Bronchoscopy revealed a broncholith in B4. Component analysis showed that more than 98% of this stone consisted of calcium carbonate. Case 2 was a 75-year-old male who was hospitalized because of continuous cough. The chest radiograph showed calcification and atelectasis in the right upper lobe. Bronchoscopically, right B3 was obstructed by a broncholith. After removal of the stone, the distal part of B3 was noted to be filled with pus. Analysis of the stone's composition revealed calcium phosphate (77%) and calcium carbonate (23%).

Adult↗

Structure and expression during development of Drosophila melanogaster gene for DNA polymerase alpha.

The Drosophila melanogaster gene and cDNA which span the entire open reading frame for DNA polymerase alpha, were cloned, and their nucleotide sequences were determined. The gene consists of 6 exons separated by 5 short introns. The major transcription initiation site was localized 85 bp upstream from the initiation codon. The nucleotide sequence of the open reading frame revealed a polypeptide of 1,505 amino acid residues with a molecular weight of 170,796. The amino acid sequence of the polypeptide was 37% homologous with that of the catalytic subunit of human DNA polymerase alpha. This sequence contains six regions, the orders and amino acid sequences of which are highly conserved among a number of other viral and eukaryotic DNA polymerases. We found 7 amino acid residues in the region between the 639th and 758th positions, identical to those essential for the active site of Escherichia coli DNA polymerase I-associated 3'----5' exonuclease. Thus, the exonuclease activity may be associated with Drosophila DNA polymerase alpha. Levels of the DNA polymerase alpha mRNA were high in unfertilized eggs and early embryos, relatively high in adult female flies and second-instar larva, and low in bodies at other stages of development. This feature of the expression is similar to that of the proliferating cell nuclear antigen (an auxiliary protein of DNA polymerase delta) and seems to coincide with the proportions of proliferating cells in various developmental stages. As the half life of the mRNA for DNA polymerase alpha in cultured Drosophila Kc cells was 15 min, expression of the DNA polymerase alpha gene is probably strictly regulated at the step of transcription.

Amino Acid Sequence↗

Cloning, sequence analysis and transcriptional expression of a ras gene of the edible basidiomycete Lentinus edodes.

In the edible basidiomycete, Lentinus edodes, the presence of a high level of intracellular cyclic AMP (cAMP) is closely related to the onset of fruiting and/or primordium formation. Since a close relationship between intracellular cAMP levels and expression of ras genes was reported for organisms such as Saccharomyces cerevisiae and Dictyostelium discoideum, we have cloned and sequences a ras gene homologue from L. edodes (Le.), and analyzed its expression during development of the fungus. This gene, named Le.ras, has a coding capacity of 217 amino acids (aa) interrupted by six small introns. The deduced Le.Ras protein exhibited the highest homology to the Schizosaccharomyces pombe RAS protein (219 aa): 86% homology in the N-terminal 80-aa sequence and 74% homology in the next 80 aa. The Le.ras gene was transcribed at similar levels during mycelial development in fruiting-body formation, suggesting no direct correlation of Le.ras expression with intracellular cAMP levels in this organism.

Acetylation↗

Blasticidin S-resistance gene (bsr): a novel selectable marker for mammalian cells.

Blasticidin S is a microbial antibiotic that inhibits protein synthesis in both prokaryotes and eukaryotes. The blasticidin S-resistance gene (bsr), isolated from Bacillus cereus K55-S1 strain, was inserted into pSV2 plasmid vector and introduced into cultured mammalian cells by transfection. The bsr gene was integrated into the genome and conferred blasticidin S resistance on HeLa cells. The transfection frequency of the bsr gene was as high as that of the aminoglycoside phosphotransferase gene, the so-called neo gene, which is a representative selectable marker for mammalian cells. Transfectants in which several copies of bsr had been integrated into the genome were highly resistant to blasticidin S. Furthermore, blasticidin S killed the cells more rapidly than G418, which is conventionally used as a selective drug for the neo gene. Thus bsr is concluded to be useful as a drug-resistance marker for mammalian cells.

Anti-Bacterial Agents↗

Existence of exine-free airborne allergen particles of Japanese cedar (Cryptomeria japonica) pollen.

We investigated whether exine-free pollen allergen particles exist together with the intact pollen grains of Japanese cedar (Cryptomeria japonica) in the air during the pollen season in Yamagata City. First, we separated the allergen particles in an Andersen multi-stage air-sampler according to their aerodynamic diameters. The amount of major allergen (Cry j I) on each stage of the sampler was determined by a sensitive fluorometric sandwich ELISA, and the pollen count of the same samples was done by light microscopy after Carberla staining. Cry j I was found in stages 1 to 6, whereas most of intact and ruptured pollen grains were microscopically observed only in stages 1 and 2. Second, we suctioned the air through a tandem membrane filter system (the first filter, Nuclepore filter with 5 microns-pores; and the second, Millipore filter with 0.3 micron-pores). None of the pollen grains was detectable on the 0.3 micron-pore filter with light microscopy. However, Cry j I was detectable in the aqueous extract from the second filter. From these results, we concluded that pollen-free Cry j I existed in the air of Yamagata City during the pollen season.

Air Pollutants↗

[Isolation and identification of Chlamydia spp. from effusion of otitis media].

Three chlamydial strains isolated from patients of otitis media with effusion were studied by comparing reactivity to monoclonal antibody (MAb) and polyclonal antibody (PAb) produced against one clinical isolate (named Mk), which was first isolated by Dr Mukai (Mukai Microbiological Research Laboratory, Yamato-shi, Kanagawa prefecture). Commercially supplied antibody (Microtrak (Syva), Culture-set (Ortho diagnostic system)) was also used. To isolate the Chlamydia spp, the yolk sacs of eggs were immediately inoculated with sample effusions (0.2 to 0.4 ml per sac) as soon as the samples were received. The eggs were observed every day for a period of 12 days thereafter for signs of life or death. One to two blind passages were first done in the eggs and then in HeLa 229 cells. The reactivity was examined by both micro-IF tests, among various strains of Chlamydia (C. trachomatis: L2. C. pneumoniae, C. psittaci: Budgerigar, Izawa, Meningopneumonitis (MP)) and by immunoblot analysis. Chlamydia spp were isolated in two of the twenty-nine sample effusions (6.9%). These isolates were then tested for reactivity to MAb and PAb. It was found that MAb reacted with MP and Mk, but not with Budgerigar, Izawa and C. pneumoniae. The antibody of Culture-set reacted with C. trachomatis C. pneumoniae and C. psittaci. No reactivity was observed in Mk by MicroTrak. Immunoblot analysis revealed that MAb reacted with about 95 KDa protein of Mk, the two clinically isolated Chlamydia spp and MP. By using PAb from rabbits, similar blotting patterns were observed in Mk, the clinical isolates and MP.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

[Occurrence of Escherichia coli in urinary tract infection].

Regarding P-fimbriae, K1 antigen, sero O-type and factors on the host (age, sex and morbidity) with are the pathogenic factors in E. coli, an assessment has been made as to what relations these factors have in urinary tract infection. By age, cases of P-fimbriae is seen in large numbers among pregnant women and infants but less among the people aged 16-30. A similar trend was observed regarding K1 antigen. An assessment made by sex show a trend that E. coli which carries P-fimbriae tends to emerge frequently in males. By morbidity, P-fimbriated strain was seen in many severe cases among pregnant women and infants. The fact has become evident further as a result of making an investigation with K1 antigen added. However, the result of an assessment made in adult cases showed that P-fimbriae was seen frequently in simple cystitis conversely. As the reason for P-fimbriae being observed more in males, participations of immunological stage and anatomical factors on the host side in the pathogenicity was resumed, and the main cause that P-fimbriae is seen more in severe pregnant and infantile cases was presumed to be vesicoureteral reflux. Regarding group sero O-type and morbidity, the types reported so far were detected also at our hospital, but a very interesting finding we have obtained was that the stains P-fimbriae and K1 antigen were observed in types 2, 12, 18 and 75.

ABO Blood-Group System↗

[Intra-pulmonary suture abscess with hemoptysis after partial resection--concerning to the pathogenesis of the suture abscess].

A 53-year-old male with a intra-pulmonary suture abscess was admitted with recurrent hemoptysis. Five year before this admission, he had undergone a partial resection of the left upper lobe for pneumothorax using unabsorbable sutures (braided polyester). Chest CT showed tumor shadow in the left S1(+2) a on admission. Because of recurrent hemoptysis, left upper lobectomy was performed. Hemoptysis was due to the presence of a suture abscess. Pathological and clinical analysis of the intra-pulmonary suture abscess showed that it was caused not only by foreign body reaction but also transbronchial infection.

Hemoptysis↗

Folliculosebaceous cystic hamartoma. A distinctive malformation of the skin.

Five examples of a distinctive sebaceous malformation, designated "folliculosebaceous cystic hamartoma," form the basis of this study. Criteria are proposed for specific diagnosis of this hamartoma and for differentiation of it from sebaceous gland hyperplasia, steatocystoma, dermoid cyst of skin, and sebaceous trichofolliculoma.

Adult↗

[A case of mediastinal mature teratoma with elevated of CA 19-9 in tumor fluid].

A 20-year-old female was diagnosed as mediastinal tumor and admitted to our hospital for the operation. Among the preoperative examinations, the serum level of CA 19-9 was within normal limits. The fluid inside the cystic tumor which was removed during the operation showed high levels of CA 19-9. We discussed the relationship between CA 19-9 and cystic lesions. Histopathological study revealed that CA 19-9 was produced in only epithelial cell of the cystic lesion. And we speculated that examination of serum CA 19-9 helps the diagnosis of cystic lesions.

Adult↗

Isolation of temperature-sensitive cell cycle mutants from mouse FM3A cells. Characterization of mutants with special reference to DNA replication.

A large number of mutants that are temperature sensitive (ts) for growth have been isolated from mouse mammary carcinoma FM3A cells by an improved selection method consisting of cell synchronization and short exposures to restrictive temperature. The improved method increased the efficiency of isolating DNA ts mutants, which showed a rapid decrease in DNA-synthesizing ability after temperature shift-up. Sixteen mutants isolated by this and other methods were selected for this study. Flow microfluorometric analysis of these mutants cultured at a nonpermissive temperature (39 degrees C) for 16 h indicated that five clones were arrested in the G1 to S phase of the cell cycle, six clones were in the S to G2 phase, and two clones were arrested in the G2 phase. The remaining three clones exhibited 8C DNA content after incubation at 39 degrees C for 28 h, indicating defects in mitosis or cytokinesis. These mutants were classified into 11 complementation groups. All the mutants except for those arrested in the G2 phase and those exhibiting defects in mitosis or cytokinesis showed a rapid decrease in DNA synthesis after temperature shift-up without a decrease in RNA and protein synthesis. The polyomavirus DNA cell-free replication system, which consists of polyomavirus large tumor antigen and mouse cell extracts, was used for further characterization of these DNA ts mutants. Among these ts mutants, only the tsFT20 strain, which contains heat-labile DNA polymerase alpha, was unable to support the polyomavirus DNA replication. Analysis by DNA fiber autoradiography revealed that DNA chain elongation rates of these DNA ts mutants were not changed and that the initiation of DNA replication at the origin of replicons was impaired in the mutant cells.

Animals↗

Cerebral circulation and oxygen metabolism associated with subclinical periventricular hyperintensity as shown by magnetic resonance imaging.

A combined magnetic resonance imaging and positron emission tomography study was performed on 21 patients with cerebrovascular risk factors but without neurological abnormalities. Our purpose was to investigate the hypothesis that periventricular hyperintensity (PVH) reflects ischemia. Periventricular hyperintensity was evaluated with a method we devised, and cerebral circulation and oxygen metabolism were evaluated with the oxygen-15 steady-state technique. We concluded that the brain with severe periventricular hyperintensity had abnormal circulation, although oxygen metabolism was not measurably affected. The role of a compensation mechanism under conditions of decreased oxygen supply was considered.

Aged↗

Cutaneous T cell lymphoma differing from classical mycosis fungoides and Sézary syndrome: clinical, histological and immunohistochemical studies of six cases.

Six cases of cutaneous T cell lymphoma (CTCL), differing from classical mycosis fungoides (MF) and Sézary syndrome (SS), were studied. Initial skin eruptions were erythematoplaques, papules and tumors. Some cases showed frequent spontaneous regressions without any specific therapy; however, the prognosis was not always good. Surface phenotypes of neoplastic cells were markedly heterogeneous from the onset of initial skin eruption. Three cases expressed the T helper/inducer (Th/i) phenotype, one case had both the Th/i and T suppressor/cytotoxic phenotypes, and the other two cases lacked both surface markers. These results suggest that CTCL other than MF and SS displays marked heterogeneous clinical behavior and diverse morphological and immunophenotypical characteristics of the postthymic T cells.

Aged↗

[Evaluation of in vitro activity of rokitamycin against Chlamydia trachomatis].

Evaluation of the in vitro activity of rokitamycin (RKM) against Chlamydia trachomatis in cycloheximide treated HeLa 229 cells and McCoy cells by comparing with five drugs including doxycycline (DOXY), minocycline (MINO), ofloxacin (OFLX), ampicillin (ABPC) and erythromycin (EM) with regard to assaying minimal inhibitory concentrations (MICs), minimal lethal concentrations (MLCs), and by yield reduction assays: 1) direct treatment of Chlamydial organisms with various concentrations of antibiotics before inoculation, 2) pre-treatment of host cell (HeLa 229) with the antibiotics before they are infected and 3) treatment of already infected cultures (48 hrs after infection) with antibiotics. The yield of Chlamydia was determined by both assaying the infectivity of Chlamydia and/or Chlamydiazyme value (from Abott Co Ltd USA). It was found that similar MIC was obtained among the drugs tested (except EM) in both HeLa 229 cell and McCoy cell assay system. The MLC of RKM (0.3 micrograms/ml) was the same as that of OFLX and was significantly lower than that of other drugs tested. When Chlamydial organisms and the host cells were treated with various concentrations (25-0.1 micrograms/ml) of the drug, the infectivity and the growth of Chlamydia was noteworthily decreased with RKM treatment. Infectivity of Chlamydia in an already infected cultures also decreased with RKM treatment within 24 hours when comparing the value of the control. In other drugs treatment, 96 hours or more hours were required for obtaining the same infectivity as RKM.(ABSTRACT TRUNCATED AT 250 WORDS)

Cells, Cultured↗