Allelic variations in the TAP2 and LMP2 genes in Behçet's disease.
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ATP Binding Cassette Transporter, Subfamily B, Mem↗
Biomedical subjects
S Ohno
Publications and source records attributed to S Ohno.
A significant association of HLA-DPB1*0501 with Vogt-Koyanagi-Harada's disease results from a linkage disequilibrium with the primarily associated allele, DRB1*0405.
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Adult↗
Genetic polymorphisms in the keratin-like S gene within the human major histocompatibility complex and association analysis on the susceptibility to psoriasis vulgaris.
Psoriasis vulgaris is associated with the HLA-Cw6 and Cw7 antigens. However, it has not yet been clarified if the HLA-Cw6 and Cw7 genes themselves are the susceptible gene related to this disease or if it is some other non-HLA gene in a linkage disequilibrium with these HLA-C alleles. The S gene, recently identified in the HLA class I region 160 kb telomeric of HLA-C, encodes a keratin-like protein and is expressed specifically in the granular layer of the epidermis. Therefore, it is tempting to speculate that the S gene is one of the strong candidate genes responsible for the pathogenesis of psoriasis vulgaris. Direct sequencing of the first and second exon of the S gene after polymerase chain reaction (PCR) amplification has allowed the identification of two diallelic polymorphic sites in exon I and seven diallelic polymorphic sites in exon 2, three among which result in amino acid exchanges, a Ser-Phe substitution at amino acid position 186, a Gly-Val substitution at position 393 and a Ser-Leu substitution at position 394. No significant difference in the dimorphic distributions of the S gene was observed between the patients with psoriasis vulgaris and healthy controls, suggesting that the susceptible gene for psoriasis is not the S gene itself.
Disease Susceptibility↗
Mesial temporal lobe epilepsy in childhood.
To clarify the clinical picture of mesial temporal lobe epilepsy (MTLE) in childhood, we carried out a clinical, electroencephalographic, and neuroradiologic study of 19 patients. MTLE was noted in 19 (0.82%) of 2,319 epileptic patients with childhood onset. Three types of initial seizure were recognized: febrile convulsion, afebrile generalized convulsion, and complex partial seizure (CPS). As presumed causes, various prolonged convulsions (persisting for > 30 min) were found in 12 (63.2%) cases. Regardless of the presence of preceding convulsions (febrile or afebrile), the clinical course was not uniform, with CPS in the early period temporarily controlled in some cases and intractable from the early period in others. Unilateral hippocampal abnormalities were confirmed on magnetic resonance imaging (MRI) before the age of 5 years in two cases, suggesting that mesial temporal sclerosis (MTS) is formed within a relatively short period in some cases. Seizures were controlled for > 6 months in only two (10.5%) cases and persisted in 17. In four (21.1%) cases, surgical treatment was considered to be available.
Age Factors↗
Genome analysis of adenovirus type 3 isolated in Japan.
Adenovirus type 3 (Ad3) isolates, isolated from 45 patients with acute conjunctivitis during the year 1990 in Japan, were studied by DNA restriction enzyme analysis with restriction endonucleases recognizing 6-bp sequences (BamHI, SmaI, HindIII, BglII) and endonucleases recognizing 5- or 4-bp sequences (HinfI and TaqI). All 45 isolates of Ad3 were identified as the genome type Ad3f by six endonucleases. They were further classified into three varieties by HinfI, varieties H1 (87.5%), H2 (8.9%), and H3 (2.2%), and into five varieties by TaqI, varieties T1 (75.6%), T2 (13.3%), T3 (2.2%), T4 (4.5%), and T5 (8.9%). The use of HinfI and TaqI was sufficient to distinguish six subgenome types: types Ad3fH1T1, Ad3fH1T2, Ad3fH1T4, Ad3fH1T5, Ad3fH2T1, and Ad3fH3T3. Among them, Ad3fH1T1 was predominant in areas where the virus is epidemic. The study showed that the same Ad3 genome type, genome type Ad3f, caused acute viral conjunctivitis even in different areas in Japan. The source and the route of infection were suggested to be common in areas where the virus is epidemic.
Adenovirus Infections, Human↗
Rapid diagnosis of adenoviral conjunctivitis by PCR and restriction fragment length polymorphism analysis.
To detect and identify adenovirus (Ad), we used a combination of PCR and restriction fragment length polymorphism (RFLP) analysis. Nested PCR with two primer sets that hybridize to the conserved region for hexon proteins of 14 prototypes of Ad, Ad serotype 1 (Ad1) to Ad8, -11, -14, -19, -37, -40, and -41, amplified a 956-bp DNA fragment. The amplified fragments from the 14 prototypes were completely differentiated with a combination of three restriction endonucleases, EcoT14I, HaeIII, and HintI. We applied this new method for 127 samples of conjunctival scrapings from patients with conjunctivitis and compared the results with those obtained with the combination of culture isolation and a neutralization test (NT). PCR gave a positive result in 69 of 127 cases (54.3%), while only 61 of the 127 samples (48.0%) tested positive by culture isolation. Compared with isolation, the PCR method had a sensitivity of 100% (61 of 61). Positive PCR samples were further classified as Ad37 (59.5%), -3(31.9%), -11 (4.3%), -8 (2.9%), and -4 (1.4%) by PCR-RFLP analysis. Of eight samples that were PCR positive and culture isolation negative, six were Ad37 and two were Ad8 by PCR-RFLP analysis. These differentiations of isolation-positive samples were identical to the results obtained by the NT. It took only 3 days to detect and identify Ad by PCR-RFLP analysis, whereas it took at least 3 weeks by culture isolation and NT. Our newly developed method of detecting and typing human Ad by PCR-RFLP analysis is more sensitive, accurate, and rapid than the conventional method of culture isolation and an NT.
Adenoviruses, Human↗
Proteoglycans in articular cartilage revealed with a quick freezing and deep etching method.
OBJECTIVES: To clarify the three dimensional ultrastructure of proteoglycans, and their relationship with other matrix components in articular cartilage. METHODS: Specimens from rat femoral heads were examined using three techniques: (1) Histochemical staining with cationic polyethyleneimine (PEI), using a pre-embedding or a postembedding method. Some tissues were pretreated with chondroitinase ABC or hyaluronidase. (2) Quick freezing and deep etching (QF-DE). Some specimens were fixed with paraformaldehyde and washed in buffer solution before quick freezing; others were frozen directly. (3) Ultrathin sections were studied after conventional preparation. RESULTS: Proteoglycans were observed as aggregated clumps with PEI staining by the pre-embedding method, but as fine filaments by the postembedding method. They were lost with enzyme digestion; this was also demonstrated by the QF-DE method. The ultrastructure was well preserved by the QF-DE method when fixation and washing procedures were included, but not without these procedures. A fine mesh-like structure was connected to the cell membrane in the pericellular matrix. Filamentous structures suggestive of aggrecans were observed among collagen fibrils. They had side chains, approximately 50 nm in length, which branched from the central filaments at intervals of 10-20 nm, and were occasionally linked to other structures. Many thin filaments were also attached to the collagen fibrils. CONCLUSIONS: The QF-DE method incorporating paraformaldehyde fixation and buffer washing procedures revealed three dimensional, extended structures suggestive of proteoglycans.
Animals↗
Insufficient expression of Fas antigen on helper T cells in Behçet's disease.
AIMS: To investigate the lack of equilibrium in the regulatory mechanism of the immune system in Behçet's disease (BD), the expression of Fas antigen, an apoptosis related antigen, on peripheral blood lymphocytes from BD patients was analysed. METHODS: Twenty one BD patients were the subjects in this study. Ten healthy adults were examined as controls. Cell surface antigens of lymphocytes were analysed with flow cytometry. RESULTS: There was a significant (p < 0.01) difference in the proportion of CD4 positive cells with CD25 between BD patients with active uveoretinitis (27.6% (SD 8.4%)) and the controls (14.7% (2.3%)), but no significant difference in the proportion of CD4 or CD45RO positive cells with Fas. On the other hand, the proportion of CD8 positive cells with Fas was significantly (p < 0.01) higher in BD patients with active uveoretinitis (45.6% (11.6%)) than in those with inactive uveoretinitis (23.8% (8.1%)) or in the controls (24.4% (2.5%)). The proportion of CD19 positive cells with Fas was also significantly (p < 0.01) higher in BD patients with active uveoretinitis (13.0% (5.0%)) than in the controls (5.1% (2.1%)). CONCLUSION: The insufficient expression of Fas on activated CD4 positive T cells and its high expression on CD8 positive T cells seem to play an important role in the chronic inflammation in BD.
Adult↗
Breast lesions: correlation of contrast medium enhancement patterns on MR images with histopathologic findings and tumor angiogenesis.
PURPOSE: To compare qualitative and quantitative magnetic resonance (MR) mammographic features of breast lesions with histopathologic findings, especially tumor angiogenesis. MATERIALS AND METHODS: Seventy-three patients (72 women, one man; aged 30-78 years; mean age, 51.0 years) with suspicious breast lesions underwent MR imaging. Noncontrast medium-enhanced localization imaging and then gadolinium-enhanced dynamic fast spoiled gradient-recalled-echo (SPGR) imaging were performed in all patients. In selected patients, subtraction fast SPGR images were obtained. The Pearson and Spearman correlation tests were used to determine the strength of the relationships between enhancement parameters and microvessel determinations. RESULTS: Time intensity curve type correlated with microvessel density grade (Spearman rank correlation test: r = .90, P < .001). The steepest slope of contrast medium uptake correlated with microvessel counts (Pearson correlation test; r = .83, P < .001). Peripheral enhancement in invasive carcinomas (n = 9) correlated with high peripheral and low central microvessel densities, which were associated with desmoplasia and/or necrosis. Internal septations (n = 2) were seen only in fibroadenomas. CONCLUSION: The density and distribution of microvessels may play major roles in the determination of the initial rate of contrast medium uptake and the heterogeneity of tumor enhancement.
Adult↗
Ultrastructural study of the cytoskeleton of optic nerve axons in guinea pigs as revealed by a quick-freezing, deep-etching method.
The ultrastructure of the cytoskeleton of optic nerve axons in guinea pigs was examined by the quick-freezing and deep-etching (QF-DE) method. The optic nerve tissues were treated with 0.5% saponin before QF. In the replicas, the axoplasm, as observed with conventional ultrathin sections, was seen to be composed of longitudinally oriented microtubules (MT) and neurofilaments (NF). Thin and elaborate cross-linking structures were observed in the interstices between MTs, NFs, and membranous organelles. They consisted of two different types: NF-associated cross-linking structures (about 20-50 nm in length) and MT-associated structures (about 10-20 nm in length), and may play a role in the slow transport of NF and MT and fast transport of organelles, respectively.
Animals↗
Clinical and immunogenetic investigation of a Laotian patient with Vogt-Koyanagi-Harada's disease.
Vogt-Koyanagi-Harada's (VKH) disease has been known to be associated with HLA class II antigen, HLA-DR4 and -DR53. Recent recombinant DNA technical advances have allowed us to investigate the genetic polymorphism of HLA antigens at the molecular level. By use of the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique we have found that VKH disease is closely associated with HLA-DRB1*04 (DRB1*0405/ DRB1*0410) and DQB1*04 (DQB1*0401/DQB1*0402) in the Japanese. Here, we will describe a typical case of a Laotian patient with VKH disease and the results of HLA typing: the patient showed a conservation of one of the major haplotypes observed in Japanese VKH patients, DRB1*0405-DQB1*0402.
Adult↗
A carboxy-terminal deletion mutant of protein kinase C beta II inhibits insulin-stimulated 2-deoxyglucose uptake in L6 rat skeletal muscle cells.
Alternative splicing of pre-mRNA encoding the carboxy-terminal (C-terminal) exons of protein kinase C beta (PKC beta) leads to the expression of two protein isoforms, PKC beta 1 and PKC beta II, with the potential for different functions. PKC beta II expression is regulated by insulin via alternative mRNA splicing. A physiological consequence of its activation was investigated in L6 rat skeletal muscle cells expressing GLUT4 transporters, a cell line in which PKC is involved in glucose transport. We examined the contribution of PKC beta II for insulin-stimulated [3H]2-deoxyglucose uptake by constructing three PKC beta II C-terminal deletion mutants designated M216, M217, and M218. When transiently expressed in COS1 cells, M217, with nine amino acids deleted, demonstrated autophosphorylation activity 10-fold less than full-length PKC beta II. The mutants M218, with 13 amino acids deleted, and M216, with 52 amino acids deleted, demonstrated no autophosphorylation activity and are kinase negative. When transiently expressed in L6 myotubes, M217 inhibited insulin-stimulated 2-deoxyglucose uptake by 45% (with a 45% transfection efficiency) whereas M216 and M218, kinase-negative mutants, had no effect compared with cells transfected with control plasmid. Cotransfection of full-length PKC beta II with M217 was able to rescue the inhibition of insulin-stimulated 2-deoxyglucose uptake as compared with cotransfection of M217 with the control plasmid, suggesting that M217 acts as a dominant-negative. In contrast, cotransfection of full-length PKC beta I, the other alternatively spliced form, did not rescue inhibition of insulin-stimulated 2-deoxyglucose uptake by M217. To further demonstrate the involvement of PKC, specifically PKC beta II, in insulin-stimulated 2-deoxyglucose uptake, we used two inhibitors, CG41251 (a specific PKC inhibitor) and CG53353 (a PKC beta II-specific inhibitor at 1 microM). Both inhibited insulin-stimulated 2-deoxyglucose uptake 50-60% in L6 myotubes. We conclude that M217 may act as a specific PKC beta II dominant-negative and that PKC beta II is more specific for insulin-stimulated 2-deoxyglucose uptake in these cells than PKC beta I.
Amino Acid Sequence↗
[Advances in the treatment of colorectal cancer].
Colorectal cancer is one of the most serious diseases of our time. Recent progress in its treatment is reviewed in this paper. There are many approaches for treating colorectal cancer; extended surgery to function-preserving surgery. The aim of surgical treatment is to excise the tumor completely, so extended surgery has been performed for curative resection of colorectal cancer. However, the quality of life (QOL) of the patients has been extremely impaired. In recent years, surgical treatments for avoiding colostomy, and urinary and sexual difficulties have been widely carried out, and QOL of the patients could be maintained in the postoperative period. Stage-oriented therapy is accepted as the concept of surgical treatment of colorectal cancer. For further improving the clinical results of colorectal cancer, we advocated "type-oriented therapy" which is based on the biological characteristics of colorectal cancer, by determining i.e., p53 abnormalities of cancer cells, serum p53 antibodies, micrometastasis in the bone marrow and/or sensitivity to anticancer drugs.
Antineoplastic Combined Chemotherapy Protocols↗
[Ultrastructural study on rat precorneal tear film by quick freezing-freeze substitution method].
The ultrastructure of rat precorneal tear film was visualized by the quick freezing-freeze substitution (QF-FS) method and the conventional fixative method under the scanning electron microscope (SEM). In the QF-FS method, the eyeballs of rats were quickly frozen with an isopentane-propane mixture cooled by liquid nitrogen applied directly to the eyes. After enucleating the eyes and fracturing the frozen corneas, the corneas were prepared for SEM observation. In contrast to the conventional fixative method, by which the microvilli of the surface epithelial cells could be visualized clearly, a very thin membrane-like structure was observed to cover the corneal surface in the eyes prepared by the QF-Fs method. Between the membrane-like structure and corneal surface, a homogeneous and fine network-like structure was observed. The results suggest that the structure of the tear film might be different from the one we have believed to consist of three layers until now. The QF-FS method is considered to be useful for the morphological study of the precorneal tear film.
Animals↗
[An aspect of the research on protein kinase C].
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Amino Acid Sequence↗
Protein kinase C beta 1 and protein kinase C beta 2 activate p57 mitogen-activated protein kinase and block differentiation in colon carcinoma cells.
When HD3 colon carcinoma cells differentiate to fluid-transporting, enterocytic-like cells, they down-regulate their protein kinase C (PKC) beta levels 5-10-fold and lose two responses to basic fibroblast growth factor (FGF): proliferation and the ability to activate p57 mitogen-activated protein (MAP) kinase. HD3 cells were transfected with expression plasmids for the splice variants PKC-beta 1 and PKC-beta 2 and the empty vector for a control. Each of two PKC-beta 1 and each of two PKC-beta 2 transfectant clones exhibited elevated levels of Ca(2+)-and phosphatidylserine-dependent PKC activity. Both PKC-beta 1 transfectant clones had elevated levels of PKC-beta 1 protein compared with the PKC-beta 2 transfectants or controls, whereas both PKC-beta 2 transfectant clones had elevated levels of PKC-beta 2 protein compared with PKC-beta 1 transfectants. Control transfectants had no detectable PKC-beta 2 protein. Similar levels of PKC-alpha were found in all lines. Each PKC-beta transfectant was less differentiated than the parental line and had regained proliferative response to basic FGF. Increased growth rates in athymic mice were seen for PKC-beta 2 and PKC-beta 1 transfectant cells. Immunocytochemistry of the sectioned tumors showed enhanced protein levels of PKC-beta 2 and PKC-beta 1, correlating increased levels of these isonzymes with increased growth. Increased myelin-basic protein (MBP) kinase activities of M(r) 44,000, 57,000, 63,000, 110,000, and 130,000 by in-gel kinase assay characterized each PKC-beta transfectant. Both Western blotting and immunoprecipitation studies from 35S-prelabeled cells with a pan-erk antibody showed no increase in protein abundance of MAP kinases of M(r) 44,000, 57,000, and 63,000, suggesting that elevated PKC-beta levels led to activation of the smaller three MAP and MBP kinases. Activation of p57 MAP kinase in each PKC-beta transfectant was demonstrated by immunoprecipitation with an antiphosphotyrosine monoclonal antibody and then by assay of the immunoprecipitates by in-gel kinase assay on MBP. p57 MAP kinase was distinguished from the M(r) 54,000 stress-activated protein kinases, which migrated more rapidly on SDS gels and could be detected by in-gel kinase assay on MBP only after cellular stress. Thus, expression of elevated levels of PKC-beta 1 and PKC-beta 2 in differentiated HD3 colon carcinoma cells blocked their differentiation, enabled them to proliferate in response to basic FGF like undifferentiated cells, increased their growth rate in athymic mice, and activated several MBP kinases, among them, p57 MAP kinase.
Alternative Splicing↗
HLA serological and class II genotyping in sarcoidosis patients in Japan.
To investigate the HLA alleles that contribute to the genetic susceptibility to sarcoidosis, HLA serological typing was performed in 75 patients with sarcoidosis and 150 controls using the standard complement-dependent microcytotoxicity method. The genomic DNAs of the 75 patients and 130 of the 150 controls were used to analyze HLA-DRB1, -DQA1, -DQB1 and -DPB1 alleles, utilizing the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Serological typing showed that the frequencies of HLA-DR52, -DR5, -DR6 and -DR8 were significantly increased in the patients compared to the controls. In PCR-RFLP genotyping, the frequencies of the DR52-associated DRB1 alleles (DRB1*11, DRB1*14), DRB1*08, DQA1*0501 and DQB1*0301 were significantly increased in the patients compared to the controls. The frequencies of the DRB1*12 alleles were also increased among the patients, but this increase was not significant. The frequencies of DRB1*0101, DQB1*0501, and DPB*0402 were significantly lower in the patients than in the controls. The significant increases in the frequencies of DQA1*0501 and DQB1*0301 could be due to the linkage disequilibrium between the DR52-associated DRB1 alleles and DQA1*0501 and DQB1*0301 alleles among the Japanese. The significantly increased frequency of the DR8 (DRB1*08) haplotype, which lacks the DRB3 gene encoding DR52 antigen, suggested that the DR5 (DRB1*11), DR6 (DRB1*14) and DR8 (DRB1*08) of the DRB1 alleles may determine the susceptibility to sarcoidosis among the Japanese.
Adolescent↗
An ultrastructural study of goblet cells in rat nasal mucosa as revealed by the quick-freezing method.
In order to clarify the natural ultrastructure of goblet cells in the rat nasal mucosa, they were examined by the quick-freezing and freeze-substitution (QF-FS) or deep-etching (QF-DE) methods for comparison with conventional fixation methods. Some nasal mucosal tissues were unstimulated; others were stimulated with acetylcholine or substance P. The QF-FS method yielded fewer artefacts on transmission electron microscopy than conventional fixation methods. In the stimulated goblet cells, most of the secretory granules appeared to be loose in the matrix and more distorted in shape. By the QF-DE method, they were observed 3-dimensionally to be larger in size and aggregated together. In contrast, the secretory granules in the unstimulated goblet cells were mostly round and small, and separate from each other. It is concluded that the ultrastructure of secretory granules is artefactually modified by conventional fixation methods and that granule structure in goblet cells alters during the secretory process.
Acetylcholine↗