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Biomedical subjects

Y Fukazawa

Publications and source records attributed to Y Fukazawa.

At least 19 recordsLinked to original sources

Apoptosis in thyroid tissue from patients with Hashimoto's thyroiditis.

To clarify whether apoptosis of thyroid follicular epithelial cells occurs at the tissue level in autoimmune thyroiditis, 17 specimens of thyroid tissues with Hashimoto's thyroiditis were stained for fragmented DNA. Almost all nuclei of follicular epithelial cells forming atrophic thyroid follicles surrounded by mononuclear cell infiltration and fibrosis showed positive staining. With increasing distance from lymphoid cell follicles, the percentage of follicular epithelial cells with DNA fragmentation-positive nuclei decreased (30-80%). Electron microscopic study revealed the existence of epithelial cells with shrunk and condensed nuclei. The frequency of those cells in different areas was almost compatible with that of cells with fragmentation-positive nuclei. These findings suggest that apoptosis plays an important role in the thyroid tissue injury in autoimmune thyroiditis.

Adult

A new biomaterial, hen egg shell membrane, to eliminate heavy metal ion from their dilute waste solution.

The egg shell membrane (ESM) is an intricate lattice network of stable and water-insoluble fibers with high surface area. ESM accumulates and eliminates various heavy metal ions from dilute aqueous solution with high affinity and in short contact time, depending on pH and characteristics of the individual ion. Under certain conditions, the level of precious ions, Au, Pt, and Pd accumulation approaches 55, 25, and 22% of dry wt of ESM, respectively. Also uranium uptake 30% of that of ESM. Experiments suggested that ESM is promising to use for the purpose of removal/recovery of metals and water pollution control.

Adsorption

Antigen-specific suppression of antibody responses by T lymphocytes cytotoxic for antigen-presenting cells.

In this study we demonstrated an alternative model of the antigen (Ag)-specific suppression of antibody response in mice. Splenocytes that were taken from BALB/c mice immunized by i.v. injection of soluble human serum albumin (HSA) or ovalbumin exhibited MHC-restricted Ag-specific cytotoxicity for the respective antigen-presenting cells (APC). When HSA-primed splenocytes cultured with Ag and interleukin-2 (IL-2) were treated with anti-CD4 or anti-CD8 monoclonal antibody (mAb) plus complement, CD8+ and CD4+ T cells exhibited nearly the same level of cytotoxicity against APC. Furthermore, HSA-primed CD4+ and CD8+ T cells released the same amount of interferon-gamma (IFN-gamma) when stimulated with Ag and IL-2. Recombinant IFN-gamma was shown to suppress the in vitro plaque-forming cell (PFC) response to sheep red blood cells (SRBC) only when it was added within 24 h after addition of Ag. The supernatants from both HSA-primed CD4+ and CD8+ T cells suppressed the PFC response to SRBC in vitro, and the suppressive activity was abrogated by anti-IFN-gamma mAb, but increased by anti-IL 4 mAb. These results suggest that in our system the effector cells for Ag-specific suppression of the antibody response in mice are both the cytotoxic type 1 clones (IFN-gamma-producing) of CD4+ and CD8+ T cells for APC, and that IFN-gamma is a major extracellular effector molecule for such suppression.

Animals

[Mucinous adenocarcinoma of the prostate. A case report and analysis of the literature].

An 80-year-old man, who had been treated for colon cancer 25 years ago, presented with gross hematuria. Rectal examination revealed a soft nodule in the right lobe. The serum prostatic specific antigen (PSA) was elevated to 5.2 ng/ml, while prostatic acid phosphate (PAP) was normal. Transrectal ultrasound revealed a hypoechoic mass in peripheral zone of the prostate and dilated seminal vesicle. A needle biopsy of the prostate showed mucinous adenocarcinoma. Under the diagnosis of prostatic tumor with seminal vesicle involvement, radical prostatectomy was performed. Histological findings showed organ confined cancer, of which most was composed of extracellular mucin lakes. Immunohistochemical study revealed the tumor cells positive for PSA and PAP. Mucinous adenocarcinoma of the prostate has been known to be clinically different from non-mucinous adenocarcinoma, in that the former is insensitive to hormonal therapy, is rarely associated with elevated PAP and rarely metastasize to the bone. But our analysis of the literatures is Japan showed no significant difference clinically between mucinous and non mucinous prostatic adenocarcinoma. However mucinous adenocarcinoma with signet ring cell rarely responds to hormonal therapy, which should not be classified to true mucinous adenocarcinoma in the current criteria. True mucinous adenocarcinoma could be a variant of prostatic adenocarcinoma, which is peripheral origin and should be treated like non-mucinous adenocarcinoma.

Acid Phosphatase

Scanning electron microscope observation of adherence of Candida albicans to cultured keratinocytes.

The role of antigen 6 in the adherence process of Candida albicans serotype A to cultured keratinocytes was examined with a scanning electron microscope. The number of adhered organisms was significantly lower for the antigen 6-deficient mutant strain than for the antigen 6-positive parent strain (P < 0.001). Fibril- or strand-like structures bridging the organisms and the keratinocytes were found to develop during the later stages of adherence.

Bacterial Adhesion

New method for detection of Candida albicans in human blood by polymerase chain reaction.

We developed a new method for detection of Candida albicans cells in human blood by the polymerase chain reaction that amplifies a 125-bp region within a species-specific, multiple-copy DNA fragment. In preparing template DNA from C. albicans-containing blood, the addition of C. tropicalis cells, together with antibody which reacts with both cells, raised the sensitivity of detection of C. albicans to about 3 cells per 0.1 ml.

Base Sequence

Expression of myeloid cell phenotypes by a novel adult T-cell leukemia/lymphoma cell line.

BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) can infect a number of cells of different lineages in vitro, yet the immunophenotypes of most adult T-cell leukemia/lymphomas (ATLs) are restricted to CD4+. The apparent discrepancy between these findings is still largely unknown. PURPOSE: We report on a unique case of ATL in which the leukemia cells were positive for both T-cell and myeloid cell antigens. To characterize these cells, we isolated cell lines from this patient with ATL. METHODS: The fresh leukemia cells were cultured without the addition of interleukin-2. Cell cloning was carried out by limiting dilution. RESULTS: A cell line (MU) and its clonal sublines were established. MU cells showed the same chromosomal abnormalities and T-cell receptor beta-chain gene rearrangement pattern as those of fresh leukemia cells. MU cells were exclusively positive for a myeloid cell marker (CD13) but not for T-cell markers, despite the presence of T-cell receptor gene rearrangement. CONCLUSION: The established ATL cell line showed both T-cell and myeloid cell characteristics, which seems to be the first evidence for the close association of ATL cells with both lymphoid and myeloid features. The cell line may provide a new insight for the targets of HTLV-1 infection and transformation in vivo.

Adult

Antigenic role of stress-induced catalase of Salmonella typhimurium in cell-mediated immunity.

The ability of the H2O2-induced catalase of Salmonella typhimurium to induce cell-mediated immunity against S. typhimurium infection in mice was examined. When exponentially growing cells of S. typhimurium were treated with 20 microM H2O2, the cells resisted killing by 1 mM H2O2 and showed the induction of a new species of catalase in addition to the constitutively produced one. Two molecules of catalases in S. typhimurium were isolated from mutant strains: H2O2-induced catalase (catalase II, 320 kDa), from a regulatory gene-deficient oxyR1 mutant, and constitutive catalase (catalase I, 350 kDa), from a katG gene-deleted mutant. When mice were inoculated with a sublethal dose of live cells, an intensive protective immunity (100% survival at 3 weeks) after challenge with a virulent strain associated with the delayed-type footpad hypersensitivity (DTH) reactions to both catalase I and catalase II was induced. Conversely, mice immunized with formalin-killed virulent S. typhimurium did not elicit protective immunity or DTH to either catalase. When mice were immunized with catalase I or catalase II, an enhanced protection (to a certain extent: 50% survival at 3 weeks) was induced in mice immunized with catalase II associated with DTH which did not cross-react with catalase I but not in those given catalase I. These results suggest that H2O2-induced stress proteins, including catalase II, are the dominant antigens for cell-mediated immunity in live cells of S. typhimurium and that a burst of such stress proteins in live salmonellae in phagocytes is responsible for the induction of cell-mediated immunity that is largely involved in the protection of susceptible mice against Salmonella infection.

Animals

Role of specific determinants in mannan of Candida albicans serotype A in adherence to human buccal epithelial cells.

Candida albicans serotype A (C. albicans A) possesses a specific antigen, designated antigen 6, which resides in mannans on the cell surface. To determine the role of the mannan moiety of the C. albicans cell wall in adherence to buccal epithelial cells, we used antigen 6-deficient mutants which had been isolated by screening with an agglutinating monoclonal antibody against antigen 6 (MAb-6). 1H nuclear magnetic resonance spectral analysis of the purified mannans from the mutants showed a loss of the signals related to that beta-linkage of the side chains. Moreover, acetolyzed fragments of the mutant mannans showed a decreased amount of mannohexaose and mannopentaose. The mutant yeast cells exhibited significantly reduced ability to adhere both to exfoliated buccal epithelial cells and to a human buccal cell line. A number of strains of C. albicans A, C. tropicalis, and C. glabrata, all of which bear antigen 6, showed significantly higher adherence to the cell line than did those of C. albicans serotype B, which lack antigen 6. The whole mannan from the C. albicans A parent inhibited the adherence of C. albicans A to epithelial cells dose dependently, whereas mannan from a mutant strains did not. Moreover, C. albicans A treated with MAb-6 or polyclonal factor 6 serum showed reduced adherence. A close correlation was found between adhesive ability and agglutinability with MAb-6 in the C. albicans A parent, the antigenic mutants, and their spontaneous revertants. These results suggest that so far as mannan adhesion is concerned, serotype A-specific determinants are largely involved in the mechanisms of adherence of C. albicans A to human buccal epithelial cells.

Adhesiveness

Isolation and characterization of a species-specific DNA fragment for detection of Candida albicans by polymerase chain reaction.

A 2-kbp DNA fragment, EO3, that was present in multiple copies in the Candida albicans genome was isolated for use in developing a detection method for C. albicans by polymerase chain reaction (PCR). Dot blot hybridization revealed that EO3 was specific for the 40 isolates of C. albicans serotypes A and B used. Using a set of primers (20-mer each) derived from the nucleotide sequence of EO3, we performed specific amplification of a 1.8-kbp DNA fragment within EO3 by PCR. All 40 isolates belonging to C. albicans serotypes A and B contained amplifiable 1.8-bkp fragments, although the DNA of the amplified products exhibited small variations in size, yielding three different fragment groups. Southern blot hybridization probed with EO3 showed that these 1.8-kbp fragments were derived from the EO3 region. Conversely, the 1.8-kbp fragment was not amplified from 38 isolates belonging to seven other medically important Candida species or from isolates of Cryptococcus neoformans, Saccharomyces cerevisiae, various bacteria, and a human cell line. The detection limit of the PCR assay for C. albicans with the EO3 fragment was shown to be approximately 2 to 10 cells and 100 cells in saline and human urine, respectively, by ethidium bromide staining and 2 and 10 cells, respectively, by Southern blot analysis. In addition, EO3 was assumed to originate from mitochondrial DNA on the basis of the results of its characterizations. These results indicate that the PCR system using the 1.8-kbp fragment as a target is a reliable method for identifying C. albicans isolates, thereby suggesting its potentials for specific and sensitive detection of C. albicans in samples from patients with candidiasis.

Base Sequence

Bartter's syndrome with gouty arthritis.

A 45-year-old woman with Bartter's syndrome complicated with gouty arthritis is reported. Bartter's syndrome was diagnosed from hypokalemia, metabolic alkalosis, high plasma renin activity, normal blood pressure and attenuated pressor response to exogenous angiotensin II infusion. Serum uric acid was 11.8 mg/dl, the renal clearance of uric acid was low (3.0 ml/min) and fractional excretion of uric acid (FEUA) was 7.0%, being the lower limit of the normal range. Fractional excretion of phosphate was low (5.3%) and fractional free water clearance per fractional distal sodium delivery was 0.51. The mechanism of hyperuricemia in this case is discussed.

Arthritis, Gouty

A case of glomerular lipidosis accompanied by familial combined hyperlipidemia and panhypopituitarism.

This is a report of a case of glomerular lipidosis with familial combined hyperlipidemia and panhypopituitarism. A 60-yr-old woman was admitted for evaluation of hyponatremia. Administration of hydrocortisone normalized the level of serum Na. A pituitary hormone-stimulating test and brain computed tomography revealed panhypopituitarism with an empty sella. Glucocorticoid deficiency due to secondary hypoaldosteronism was thought to have caused the Na loss. She had been treated for thyroid dysfunction and hyperlipidemia with replacement of thyroid hormone and a lipid-lowering drug. Hyperlipidemia changed from type V into IIa in 4 yr. Furthermore, one of her brothers and one of her sons were suspected to have type IV hyperlipidemia. Familial combined hyperlipidemia accompanied by secondary hypothyroidism was thought to have increased the levels of both total cholesterol and triglyceride. Two renal biopsies in 3 yr showed lipid deposits in the mesangial cells and indicated a positive correlation between the levels of serum lipids and lipid deposits in glomeruli, which suggested an important role of abnormal lipid metabolism in the progression of glomerular lipidosis.

Empty Sella Syndrome

Comparison of serological and chemical characteristics of capsular polysaccharides of Cryptococcus neoformans var. neoformans serotype A and Cryptococcus albidus var. albidus.

The antigenic formula and chemical structure of capsular polysaccharide (CPS) of Cryptococcus albidus var. albidus (C. albidus) were studied in relation to those of C. neoformans var. neoformans serotype A (C. neoformans A). The results of slide agglutination tests with factor sera and reciprocal adsorption experiments showed that antigenic formula of C. albidus was the same as that of C. neoformans A. The soluble CPSs from the two species were obtained from culture supernatants by precipitation with ethanol followed by purification by chromatography on DEAE-cellulose column. The structural analyses of such CPSs from the two species showed that the antigenic CPS fractions consisted of a backbone of alpha(1-3)-linked D-mannopyranosyl residues with a single branch of beta(1-2)-xylose or glucuronic acid, and mostly with O-acetyl groups, in which side chains and O-acetyl groups were responsible for antigenic specificity. It was found that there was a minor difference between the CPS of C. neoformans A and that of C. albidus; in the former, unsubstituted mannose residues existed in a low frequency, but in the latter none. Moreover, the 1H-nuclear magnetic resonance spectra of partially hydrolyzed acidic fragments of the two CPSs indicated that two xylose side chains were present between glucuronic acid side chains. Taken together, it was suggested that these two species of C. neoformans A and C. albidus are closely related to each other in their CPSs.

Animals

Mechanism for candidacidal activity in macrophages activated by recombinant gamma interferon.

Candidacidal activity in macrophages activated by recombinant gamma interferon was examined kinetically in relation to acidification of phagolysosomes. In resident peritoneal macrophages (PMPs) of BALB/c mice, enhanced killing activity against Candida albicans was demonstrated after incubation with 100 U of gamma interferon per ml for 24 h but not after incubation for 48 to 72 h. Conversely, increased generation of H2O2 was exhibited in PMPs incubated from 48 to 72 h but not in PMPs incubated for 24 h. In normal PMPs, fusion of lysosomes to candida-containing phagosomes was readily accomplished and phagosome-lysosome fusion was not enhanced further by activation. The candidacidal substance was extracted from granule-rich fractions of either normal or activated PMPs by using citric acid (pH 2.7) in equal amounts; the substance showed a noncationic, heat-stable protein nature. In addition, when phagolysosomal pH was determined by flow cytometry of intraphagolysosomal fluorescein isothiocyanate-labeled C. albicans, phagolysosomes with low pH (less than 4.0) were detected in about 40% of PMPs activated for 24 h but not in those activated for 72 h or in normal PMPs. Moreover, increasing the intralysosomal pH with NH4Cl resulted in a significant reduction of candidacidal activity in activated PMPs. These results indicate that the candidacidal activity of gamma interferon-activated PMPs correlates well with enhanced acidification of their phagolysosomes and suggest that the candidacidal activity of activated PMPs is independent from reactive oxygen molecules and is mediated by proteinaceous substance(s) generated only in a strong acidic milieu of phagolysosomes by activation.

Animals

Evaluation of a new method for identification of Cryptococcus neoformans which uses serologic tests aided by selected biological tests.

A new method for identifying Cryptococcus neoformans isolates and their serotypes by the slide agglutination test using five kinds of factor sera, with the aid of nitrate reduction, phenol oxidase, and growth at 37 degrees C tests was evaluated by using 36 reference strains and 75 clinical isolates of C. neoformans. The results showed that the reference strains were identified exactly as they were labeled, and clinical isolates were identified as C. neoformans serotypes A, D, and AD. C. neoformans could be distinguished from other Cryptococcus species that cross-reacted with factor sera by their ability to grow at 37 degrees C. These results indicate that the slide agglutination test combined the use of factor sera for isolates which grow at 37 degrees C is a useful method for identification of C. neoformans and their serotypes and that the nitrate reduction test (negative in 100% of the isolates) and the phenol oxidase test (positive in approximately 95% of the isolates) can be used to confirm that the species is C. neoformans.

Agglutination Tests