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Biomedical subjects

K Komiyama

Publications and source records attributed to K Komiyama.

At least 127 records · Page 7Linked to original sources

Role of sialic acid in saliva-mediated aggregation of Pseudomonas aeruginosa isolated from cystic fibrosis patients.

The mechanism of saliva-mediated aggregation of Pseudomonas aeruginosa in subjects with and without cystic fibrosis (CF) was investigated. Virtually all saliva from CF patients that we tested strongly agglutinated the Pseudomonas cells and was heat stable to 56 degrees C, whereas saliva from subjects without CF had a decreased aggregating ability and was heat sensitive. When saliva was treated with neuraminidase and proteases, and also when P. aeruginosa cells were treated with mixed gangliosides, there was a decrease in aggregating activities. However, neither the addition of the acid-hydrolyzed ganglioside nor the treatment of the P. aeruginosa cells by sugars had any effect on subsequent aggregating activities. Therefore, the release of sialic acid by enzymatic treatments of saliva, as well as the blockage of the sialic acid-binding sites on the cell wall by mixed gangliosides, resulted in the parallel loss of saliva-mediated aggregating activity of P. aeruginosa. The level of free sialic acid released by endogenous neuraminidase was higher in the saliva from CF patients than in that from the non-CF subjects examined. The increased aggregation of P. aeruginosa mediated by saliva from patients with CF seems to be directly related to the sialic acid content present, suggesting that this acid molecule acts as the salivary receptor for P. aeruginosa.

Bacterial Adhesion↗

Interbacterial adhesion between Pseudomonas aeruginosa and indigenous oral bacteria isolated from patients with cystic fibrosis.

Interbacterial adhesion between strains of Pseudomonas aeruginosa and strains of indigenous oral bacteria, both of which were isolated from the oral cavity of cystic fibrosis patients, was investigated by the phenomenon of the coaggregation reaction. A total of 22 strains of P. aeruginosa were isolated from the oral cavity of 17 patients and examined for their abilities to coaggregate with 5 strains each of Streptococcus sanguis, Streptococcus mitis, Actinomyces viscosus, and Actinomyces naeslundii. Coaggregation reactions were common between these oral bacteria and both the mucoid and nonmucoid variants of P. aeruginosa. All strains of P. aeruginosa were also able to agglutinate neuraminidase-treated or untreated human erythrocytes of blood types A, B, and O. Positive coaggregation reactions were further characterized by determining the effects of several sugars, and of heat and protease treatments of the bacteria. None of the coaggregtion reactions were inhibited by 0.05 M lactose, galactose, glucose, fucose, or mannose. All coaggregation reactions were dependent upon heat- and protease-sensitive components of the Pseudomonas. Thus, the interbacterial adhesions between P. aeruginosa and the oral bacteria studied appears to involve adhesins on the Pseudomonas cell, which bind to complementary receptors, on the cell surfaces of oral bacteria. The apparent prevalence and diversity of interbacterial adhesions between P. aeruginosa strains originating from the oral cavity of cystic fibrosis patients and strains of the indigenous oral bacteria suggest that some of these reactions may affect the extent to which P. aeruginosa colonizes in the oral cavity of cystic fibrosis patients, and thereby, influence susceptibility of the host to infection.

Actinomyces↗

Characterization by pyocine typing and serotyping of oral and sputum strains of Pseudomonas aeruginosa isolated from cystic fibrosis patients.

Oral and sputum isolates of Pseudomonas aeruginosa in patients with cystic fibrosis were investigated. Of the 17 patients studied, 12 patients (71%) yielded both mucoid and nonmucoid variants of Pseudomonas aeruginosa from sputum and (or) various oral ecological sites, such as buccal mucosa, tongue dorsum, dental plaques, and saliva. A total of 51 strains of mucoid and nonmucoid Pseudomonas aeruginosa were isolated from these patients and were phenotypically characterized by both pyocine typing and serotyping. Five patients (42%) were colonized or infected by a single strain of Pseudomonas aeruginosa, whereas 7 patients (58%) were cocolonized or coinfected by two or more phenotypically different strains of Pseudomonas aeruginosa. To understand the mechanisms involved in Pseudomonas aeruginosa colonization, it may be necessary to identify multiple isolates of Pseudomonas aeruginosa not only from the sputum but also from the various oral ecological sites and to further explore the role of the oral cavity in this colonization.

Adolescent↗

Synergism between cellular messengers and agonist combinations in pepsinogen secretion.

The esophagus of Rana catesbeiana yielded 20-40 X 10(6) peptic cells that were greater than 80% pure by immunostaining, greater than 90% viable, and had low basal pepsinogen and lactic dehydrogenase (LDH) release. Cellular responses to agents that bypass receptors and act directly on cell messenger systems have been compared with receptor-mediated activation. Pepsinogen secretion was stimulated dose dependently by A23187, the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), and forskolin. The combination of any two or all three agents at optimum effective concentrations stimulated secretion to more than double the output of the sum of the individual responses. Incubation in Ca2+-free media reduced responses to A23187 and TPA but not forskolin. Ca2+ deprivation eliminated the synergism of messenger combinations. Secretory responses to combinations of the agonists bethanechol or bombesin with either A23187 or TPA were additive and were synergistic with forskolin. Isoproterenol plus bethanechol or bombesin produced synergistic responses that were significantly smaller than combinations of their putative messengers. The synergistic response to isoproterenol and bethanechol was dependent on extracellular Ca2+. These data suggest that although Ca2+ may function in peptic cells to stimulate secretion directly, it may also act as a gain control for synergistic interaction between other messenger pathways (protein kinase a and c).

1-Methyl-3-isobutylxanthine↗

Ultrastructural and immunocytochemical characterization of polymorphonuclear leukocytes from gingival crevice in man.

Polymorphonuclear leukocytes from the gingival crevicular fluid (CF-PMNs) of patients with generalized severe periodontitis were examined using electron microscopy and immunocytochemical techniques. CF-PMNs were found to contain numerous phagocytic vacuoles. This suggests that CF-PMNs actively phagocytized various substances from the environment. Immunocytochemical staining with FITC-conjugated IgG, IgM, and IgA reagents and TRITC-conjugated C3 reagent was applied to CF-PMNs as well as peripheral blood PMNs incubated with cell-free crevicular fluid. The cytoplasm of PMNs exhibited numerous granular foci of immunofluorescence. This finding suggests that these proteins were acquired from the environment by PMNs. The coincidental appearances of immunoglobulins and C3 in a single cell were considered to be immune complexes phagocytized by CF-PMNs in generalized severe periodontitis.

Adult↗

Antitumor effect of kazusamycin B on experimental tumors.

Kazusamycin B, a novel antibiotic (MW 542) isolated from fermentation broth of Streptomyces sp. No. 81-484 showed a broad antitumor spectrum both in vitro and in vivo. IC50 against the growth of tumor cells was around 1 ng/ml at 72 hours-exposure in vitro. Intraperitoneal injection of the antibiotic was effective in inhibiting the growth of murine tumors, S180, P388, EL-4, and B16. It was also active against doxorubicin-resistant P388, hepatic metastases of L5178Y-ML, pulmonary metastases of 3LL, and human mammary cancer MX-1 xenografted to nude mice. However, the activity of kazusamycin B toward L1210 or human lung cancer LX-1 was weaker. According to the results of comparative studies on the effect of kazusamycins B and A, an analog of B, there seemed to be no significant difference in their effectiveness. The effective dose range and toxicity were markedly dependent on tumor lines tested and the regimen used. Maximum tolerated dose in mice with subcutaneous tumors was much higher than that in mice bearing ascitic leukemia as P388. Although intermittent administration could greatly reduce the cumulative toxicity of the drug, therapeutic effect was similar with both successive and intermittent administration schedules.

Animals↗

Antitumor activity of trienomycin A on murine tumors.

The antitumor activity of a novel ansamycin antibiotic, trienomycin A, against various murine tumors was studied with two treatment schedules. The intraperitoneal injection of the antibiotic showed remarkable antitumor activity on sarcoma 180 and P388 leukemia at doses of 160 or 320 mg/kg, showing 151% and 100% increase in life span, respectively. Trienomycin A inhibited the growth of Ehrlich and Meth A cells in vitro at doses of 0.1-0.4 microgram/ml when the cells were exposed to the antibiotic for 72 hours. The incorporation of [3H]thymidine into acid precipitable material in HeLa cells was slightly more marked than that of [3H]uridine and [3H]leucine when the cells were exposed to 0.04 or 0.08 microgram/ml of trienomycin A for 4 hours. It appeared that trienomycin A showed antitumor activity by direct cytotoxic action.

Alanine↗

Kazusamycin B, a novel antitumor antibiotic.

A novel antibiotic, kazusamycin B (C32H46O7, MW 542), was isolated from the fermentation broth of Streptomyces sp. No. 81-484 and the structure was established mainly on the basis of its physico-chemical properties. Unambiguous 13C NMR spectral analysis of kazusamycin B has been also accomplished. Kazusamycin B possesses potent cytocidal activities against L1210 (IC50 0.0018 micrograms/ml) and P388 (IC100 0.0016 micrograms/ml) leukemia cells in vitro.

Animals↗

[Studies on the new antibiotic kazusamycin and related substances].

Kazusamycins A and B and leptomycin B have a structure characteristic of an unsaturated, branched-chain fatty acid with a terminal delta-lactone ring, and show antibacterial activity on some kinds of fungi. Kazusamycin A (KZM-A) showed cytotoxic activity on mammalian cells at very low concentrations (ng/ml) in vitro. The antibiotic inhibited not only the growth of transplantable murine tumors and their metastases to the lung but also human mammary tumors inoculated into nude mice. KZM-A became immediately distributed to the main organs of mice, and a certain quantity of the antibiotic was inactivated by binding to high-molecular-weight substances such as albumin. A large quantity of KZM-A was carried to the liver and excreted into the bile, but was then reabsorbed by the small intestine. The growth of tumor metastases (L5178Y cells) in the liver was suppressed by KZM-A. The antibiotic induced severe diarrhea by causing necrosis and/or lysis of the mucous membrane of the small intestine. In contrast to this, the degree of myelotoxicity was relatively slight. The active site of the fatty acid of KZM-A appeared to consist of conjugated double bonds, carboxylic acid and hydroxyl moieties.

Animals↗

Identification of an early activation antigen (Bac-1) on human B cells.

We have produced a monoclonal antibody, Bac-1, that appears to identify a novel antigen on activated human B cells. The Bac-1 antigen can be detected between 8 to 16 hr, as well as transferrin receptors (T9), after activation of small resting B cells with phorbol myristic acetate, anti-IgM antibody, Staphylococcus aureus Cowan I, or Epstein-Barr virus. The expression of the Bac-1 antigen precedes that of IL 2 receptors (Tac-1). Peak expression of the Bac-1 antigen was observed on day 3 after activation, and decreased thereafter. The Bac-1 antigen was present on a minor subpopulation of relatively large B cells isolated from blood samples, and on "preactivated" B cells of heterogeneous size isolated from spleens and tonsils. It was not detected on bone marrow pre-B cells, blood small B cells, or plasma cells, nor was it expressed by resting or activated T cells or nonlymphoid cells. Certain B cell neoplasms and B lymphoblastoid cell lines were Bac-1+, but neoplastic cells of non-B lineage were Bac-1-. With immunoperoxidase staining, Bac-1+ cells were detected predominantly in the germinal centers of tonsil sections. The Bac-1 antigen on activated B cells was destroyed by protease treatment and was enhanced by neuraminidase treatment, suggesting that the Bac-1 antibody detects a cell surface molecule via an antigenic determinant which is partially obscured by neighboring sialic acid residues. The reactivity pattern of Bac-1 differs from the patterns of cellular reactivity reported for other monoclonal antibodies with specificity for activated human B cells.

Antibodies, Monoclonal↗

Isotype-specific immunoregulation; characterization and function of Fc receptors on T-T hybridomas which produce murine IgA-binding factor.

Several methods have been used in the present study to characterize Fc receptors (FcR) expressed on T-T hybridomas derived from mouse Peyer's patch T helper (Th) cell clones that preferentially support IgA responses. These T hybridomas (designated Th HA cells) produce IgA-binding factor (IBF alpha) which regulates antigen-dependent IgA responses. The ultrastructure of Th HA cells and the distribution of Fc alpha R on these cell lines were determined by colloidal gold (CG) immunoelectron microscopy (IEM). When Th HA cells were incubated with purified mouse IgA followed by CG-labeled anti-IgA, an even pattern of CG was distributed on the cell membrane. To ensure that binding occurred through Fc alpha R, Th HA cells were mixed with MOPC 315 IgA anti-DNP, followed by staining with CG-labeled TNP-human serum albumin. This resulted in an identical pattern of gold particle distribution, confirming expression of Fc alpha R on Th HA cells. No Fc mu R or Fc gamma 1R were detectable on Th HA cells by IEM. Immunocytoadherence with TNP-conjugated erythrocytes confirmed that Th HA cells were Fc alpha R+; however, no IgM or IgG rosettes were seen. When these cell lines were analyzed by flow cytometry (FACS) using IgA, IgM, or IgG1 and FITC-labeled anti-H chain-specific antibodies, 55 to 65% of cultured Th HA cells expressed Fc alpha R, and 11 to 18% expressed Fc mu R; however, no Fc gamma 1R was detectable on Th HA cells. The use of ELISA with Th HA cells as antigen confirmed the expression of Fc alpha R and the presence of less Fc mu R on these two cell lines. Solubilized membrane fractions derived from Th HA cells were tested for the presence of FcR by ELISA and for biologic function for support of IgA responses in Peyer's patch B cell cultures. Both Fc alpha R and Fc mu R were detected in fractions derived from Th HA cells. Furthermore, these fractions supported in vitro IgA anti-sheep erythrocyte responses, comparable to those obtained with Th HA cell culture supernatants containing IBF alpha. These studies show that Th HA cells express Fc alpha R with less Fc mu R, and the solubilized form of Fc alpha R exhibits IBF alpha-like activity. The significance of FcR expression by Th cell clones and cell lines and the relationship of soluble Fc alpha R and IBF alpha for IgA response regulation are discussed.

Animals↗

Inhibition of natural killer cell activity by IgA.

The in vitro effect of IgA on natural killer (NK) activities of human peripheral blood mononuclear cells was investigated. Purified myeloma polymeric IgA2 (pIgA2) and secretory IgA (S-IgA) from human colostrum inhibited NK activity, while myeloma polymeric IgA1 (pIgA1), monomeric IgA1 (mIgA1), IgG, and IgM were ineffective. Inhibition was proportional to the concentration of pIgA2 (0.125-1 mg/ml) and was observed after as little as 1 hr of incubation at various effector to K562 target cell ratios. pIgA2 and S-IgA also inhibited NK activity of NK cell-enriched lymphoid cells and gamma-interferon-treated effector cells, but did not interfere with effector-target cell binding. The inhibitory effect was slightly diminished after 24 hr culture in pIgA2-free medium. Inhibition of cytotoxicity was not due to direct toxicity on lymphoid cells by IgA because PBL treated with pokeweed mitogen in the presence of pIgA2 or S-IgA differentiated into immunoglobulin-producing cells. Viability after 24 hr of preculture with pIgA2 and S-IgA was comparable to that of untreated control cells. Morphological examination of effector cells cultured with pIgA2 or S-IgA showed a decrease in the number of granules, and the formation of cytoplasmic vacuoles. These morphological changes appeared to coincide with the depressed cytotoxicity of NK cells. The results demonstrate that purified pIgA2 and S-IgA have significant immunomodulatory effects on human NK activity.

Binding, Competitive↗

The effect of dietary or intraperitoneally injected seaweed preparations on the growth of sarcoma-180 cells subcutaneously implanted into mice.

Sixteen preparations from 9 edible seaweeds including powdered weed (P), hot-water extract (E), the non-dialyzable fraction (I) of E and the residue (R) of hot-water extraction were incorporated into a basic diet, and they were given to mice implanted with Sarcoma-180 cells s.c., for 5 weeks. Consequently, diets with 6 preparations, E of Laminaria angustata, P and E of Laminaria angustata var. longissima, and P, E and R of Laminaria japonica var. ochotensis, were found to be effective, with inhibition ratios ranging from 70.3% to 83.6%. Intraperitoneal injection of 10 preparations from 6 edible seaweeds, including the above 3 weeds, were also noted to be effective in the same test system, with inhibition ratios ranging from 61.9% to 95.2%.

Animals↗

Glycogen synthetic abilities of Actinomyces viscosus and Actinomyces naeslundii freshly isolated from dental plaque over root surface caries lesions and non-carious sites.

Relative glycogen synthetic abilities of resting cells of fresh clinical isolates of Actinomyces viscosus and Actinomyces naeslundii originating from dental plaque samples over root surface caries lesions and non-carious sites were studied under anaerobic conditions at a constant pH of 7.0, with U-(14C)-glucose used as the carbon source. Although the rates of glucose utilization and total acid formation were essentially the same, A. viscosus strains isolated from root surface caries lesions showed glycogen synthetic abilities approximately two to seven times higher than did A. viscosus strains originating from non-carious sites, and also two to four times higher than did A. naeslundii strains originating from both carious and non-carious sites.

Acids↗

Chemical modification of herbimycin A. Synthesis and in vivo antitumor activities of halogenated and other related derivatives of herbimycin A.

Several halogenated and other related derivatives of herbimycin A have been synthesized and evaluated in vivo for their activities against Ehrlich ascites carcinoma. Some of these derivatives show higher activities than herbimycin A. Among them the derivatives modified at the 4, 5, 6, and 7-positions of the ansa chain showed particularly high activities.

Animals↗