Characterization of recombinant Streptococcus mitis-derived human platelet aggregation factor.

We previously purified Streptococcus mitis-derived human platelet aggregation factor (Sm-hPAF) from the culture supernatant of S. mitis strain Nm-65, isolated from the tooth surface of a patient with Kawasaki disease. Here we produced recombinant Sm-hPAF protein (rSm-hPAF) in Escherichia coli, to determine whether rSm-hPAF conserves its platelet aggregation activity.

[Evaluation of the GonoGen II kit for rapid identification of Neisseria gonorrhoeae using monoclonal antibody directed at gonococcal outer membrane protein 1].

The Gonogen II test for rapid identification of Neisseria gonorrhoeae (Gonococcus, GC) was evaluated. The test is based on a colorimetric reaction with monoclonal antibody to GC outer membrane protein 1. Of the 50 clinical isolates of GC, 49 isolates tested positive and only one strain tested negative.

Studies of recombinant streptococcal pyrogenic exotoxin B/cysteine protease (rSPE B/SCP) in the skin of guinea pigs & the release of histamine from cultured mast cells & basophilic leukocytes.

Background & Objectives: Streptococcal pyrogenic exotoxin B/streptococcal cysteine protease (SPE B/SCP) is considered to be one of the virulence factors of Streptococcus pyogenes (S. pyogenes) which causes serious diseases such as severe invasive infections and streptococcal toxic shock syndrome (STSS). There are no reports on the histamine releasing activity of SPE B/SCP from mast cells, although several biological activities have been studied.

Moderate hypothermia delays proinflammatory cytokine production of human peripheral blood mononuclear cells.

Objective: To clarify the influence of moderate hypothermia on the production of proinflammatory cytokines.

Design: Controlled in vitro study.

Setting: Research laboratory.

Cysteine protease activity and histamine release from the human mast cell line HMC-1 stimulated by recombinant streptococcal pyrogenic exotoxin B/streptococcal cysteine protease.

We constructed the expression vector pSK-SCP containing the streptococcal exotoxin B gene (spe b) which expressed protease activity. We showed that the recombinant streptococcal pyogenic exotoxin B/streptococcal cysteine protease (rSPE B/SCP) was secreted into the culture supernatant of the transformant and retained its SCP activity, which was equivalent to or greater than that of the naturally occurring molecule. The secreted rSPE B/SCP induced histamine release and degranulation of the human mast cell line HMC-1.

Streptococcus peroris sp. nov. and Streptococcus infantis sp. nov., new members of the Streptococcus mitis group, isolated from human clinical specimens.

Taxonomic studies were performed on eight strains of alpha-haemolytic streptococci that showed very low DNA-DNA hybridization similarity values with all established members of the mitis group of the genus Streptococcus. These strains were isolated from the tooth surface and pharynx of humans. 16S rRNA gene sequence analysis showed that these strains belonged to the mitis group, but that they fell into two new branches.

Purification and partial characterization of a novel human platelet aggregation factor in the extracellular products of Streptococcus mitis, strain Nm-65.

A human blood platelet aggregation factor was purified from the extracellular products (ECP) of Streptococcus mitis, strain Nm-65 by sequential chromatography on DEAE-Sepharose CL-6B, hydroxyapatite and Superdex 75 columns. The purified factor (S. mitis-derived human platelet aggregation factor, Sm-hPAF) gave a single band with a molecular weight of 66 kDa on SDS-polyacrylamide gel electrophoresis (SDS-PAGE).

Superantigenic exotoxin production by isolates of Staphylococcus aureus from the Kawasaki syndrome patients and age-matched control children.

Nineteen strains of Staphylococcus aureus were isolated from the throat or the tooth surfaces of 19 cases amongst 127 patients with Kawasaki syndrome (KS) during the acute phases and 11 S. aureus isolates were obtained from five of 17 diseased controls and six healthy controls. The production of exotoxins, particularly superantigenic toxic shock syndrome toxin-1 (TSST-1), coagulase serotype, pigment production, haemolytic activity and tryptophan auxotrophy of these isolates were compared.

The presence of tumor necrosis factor-alpha and its antibody in the sera of cachexic patients with gastrointestinal cancer.

Although cancer cachexia has been shown to involve several cytokines, the tumor necrosis factor-alpha (TNF) has rarely been detected in such patients. In this study, sera from 21 patients with cancer cachexia were examined for the presence of TNF and the anti-TNF antibody using an enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. All of the patients had recurrent cancer and manifested the characteristics of progressive body weight loss.

Cytokine induction by extracellular products of oral viridans group streptococci.

During an etiological study of Kawasaki disease (mucocutaneous lymph node syndrome [MCLS]), we found that dominant viridans streptococcal strains on tooth surfaces and in the throat of both MCLS patients and non-MCLS control children formed erythrogenic and biologically active, extracellular products. In this study, we demonstrated that erythrogenic culture supernatant concentrates of representative strains (two Streptococcus mitis and two Streptococcus oralis), when injected intravenously, induced serum tumor necrosis factor alpha, interleukin-6 (IL-6), and gamma interferon in muramyldipeptide- or Propionibacterium acnes-primed C3H/HeN mice. The concentrates also induced tumor necrosis factor alpha, IL-6, and thymocyte-activating factor (essentially IL-1) in murine peritoneal macrophage, human monocyte, and human whole-blood cultures.

Plasma insulin and glucagon responses to intravenous infusion of propionate and their autonomic control in sheep.

Propionate (0, 1, 2, 4, 8, 16, 32, and 64 mumol.kg BW-1 x min-1 for 30 min) was infused i.v.

Biologically active extracellular products of oral viridans streptococci and the aetiology of Kawasaki disease.

A bacteriological study of isolates from the oral cavity of patients with Kawasaki disease (KD), age-matched non-KD patients and healthy children, showed that over half the KD and control isolates had gram-positive, catalase-negative cocci. About 50% of these organisms were identified as viridans streptococci by means of an API Strep 20 kit. Further identification by fluorometric DNA-DNA hybridisation demonstrated that the predominant species were S.

[Adhesion ultrastructures of mononuclear cells in experimentally-induced silicotic granuloma].

Experimental silicosis was induced by intratracheal infusions of 1 ml saline containing 50 mg standard silica (less than 5 microns diameter) in Sprague-Dawley rats. The lung tissues were observed histologically and ultrastructurally from half an hour up to 4 months. Macrophages, neutrophils, desquamated cells and their debris piled up around the alveolar ducts where the central cores of silicotic granuloma appeared.

The adjuvant activity of pyrene in diesel exhaust on IgE antibody production in mice.

In this communication, it is shown that pyrene has an adjuvant activity on IgE antibody production when mice are immunized by an intraperitoneal injection of ovalbumin (OA) or Japanese cedar pollen allergen (JCPA) with pyrene. The effects of pyrene on IgE antibody production in mice were investigated to clarify the relation between pollen allergy and the adjuvanticity of the chemical compounds contained in diesel-exhaust particulates (DEP). In the first experiment, three groups of mice were immunized intraperitoneally six times at 2-week intervals with 1 microgram of OA alone, 1 microgram of OA plus 1 mg of pyrene, and 1 microgram of OA plus 1 mg of DEP, respectively.

Detection of antibodies against streptococcal peptidoglycan and the peptide subunit (synthetic tetra-D-alanyl-bovine serum albumin complex) in rheumatic-diseases.

Serum antibodies reactive with streptococcal cell wall peptidoglycan (PG) and its peptide subunit (synthetic tetra-D-alanine) were measured by enzyme-linked immunosorbent assay (ELISA) in patients with rheumatoid arthritis (RA), juvenile rheumatoid arthritis (JRA), osteoarthritis and acute rheumatic fever (RF) compared with healthy subjects. Using 'checkerboard' titrations, anti-PG antibody in human serum was detected at a concentration of PG antigen at 10 micrograms per well with serum dilutions of 1:1,000. For measurement of anti-tetra-D-alanine antibody, the antigen, (D-Ala4)31 was used at 0.

Immunochemical studies and complete amino acid sequence of the streptokinase from Streptococcus pyogenes (group A) M type 12 strain A374.

The complete amino acid sequence of the streptokinase (SKase) of Streptococcus pyogenes M type 12 strain A374, isolated from a patient with poststreptococcal glomerulonephritis (PSGN), was determined. The epitope domain for the monoclonal antibody N-59, which cross-reacts with SKases of both the PSGN-associated strain and S. equisimilis H46A (a non-PSGN-associated strain), was predicted to be localized in residues 370 to 374.

Detection of nephritis strain-associated streptokinase by monoclonal antibodies.

Monoclonal antibodies (MAbs) N-59 and RU-1 were produced by immunisation of mice with streptokinase secreted by Streptococcus group A, type 12, strain A374 isolated from a patient with post-streptococcal glomerulonephritis (PSGN) and were characterised by Western blot analysis. MAb N-59 recognised antigenic determinants shared by both nephritis strain-associated streptokinase (NSA-SKase) and streptokinase of Streptococcus group C (C-SKase); MAb RU-1 reacted only with NSA-SKase. All nephritis-associated group A streptococcal strains tested reacted with MAb N-59; 87.

Enzyme-linked immunosorbent assay of antibody to group A Streptococcus-specific C carbohydrate with trypsin-pronase-treated whole cells as antigen.

We describe the measurement by enzyme-linked immunosorbent assay of antibody to group A Streptococcus C carbohydrate in immunized rabbits and human sera, with trypsin-pronase-treated group A streptococcal whole cells used as the antigen. The optimal concentration of the enzyme-treated whole cells used to coat the wells was 2 x 10(7) cells per well. Rabbit antiserum diluted to 1:12,800 and human serum diluted to 1:1,000 were found to be the optimal concentrations for antibody measurement.

Streptococcal antibody: as an indicator of tonsillectomy.

The significance of antibody for streptolysin-O concerning tonsillectomy was studied. The results obtained were as follows. 1.

Coronary arteritis in mice after systemic injection of bacterial cell wall peptidoglycan.

We examined cell wall peptidoglycan (PGL) derived from group A streptococcus and other bacteria for possible induction of coronary arteritis in mouse strains. The histological finding of the main trunk of the coronary arteries of BALB/c, DBA/1J, C57BL/6 and DBA/2 mice, which were given an intravenous injection of sonicated PGL fragments of st. pyogenes at 500 micrograms per mouse 4 times at intervals of 1 week, showed diffuse cellular infiltration in the vascular wall as well as perivascular space.

Absence of nephritis strain-associated protein (NSAP) in the extracellular product of Streptococcus pyogenes strain Su.

This study was undertaken to ascertain whether or not a nephritis strain-associated protein (NSAP) is produced by Streptococcus pyogenes strain Su, which is used in OK-432, an antitumor agent. SDS-PAGE and double immunodiffusion analysis showed that no NSAP occurred in the extracellular product of S. pyogenes strain Su.