Yuko Todome

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. Subjects Peripheral blood mononuclear cells from healthy adult human subjects. Interventions Stimulation with 1 &mgr;g/mL lipopolysaccharide at 33°C and 37°C. Measurements Concentrations of released tumor necrosis factor-&agr;, interleukin-1&bgr;, and interleukin-6 were measured chronologically by enzyme immunoassay. The number of mRNA copies of these cytokines was determined by competitive reverse transcriptase-polymerase chain reaction analysis, and nuclear factor-&kgr;B activations were assessed by electrophoretic mobility shift assay. Main Results Significant reduction of the released-tumor necrosis factor-&agr; concentration was observed 1 and 2 hrs after the stimulation with lipopolysaccharide at 33°C compared with 37°C. The peak release of interleukin-1&bgr; at 33°C was delayed 12 hrs later than that at 37°C. A delayed peak in the release of interleukin-6 also was observed at 33°C. The peaks of cytokines were confirmed at the mRNA expression level by competitive reverse transcriptase-polymerase chain reaction analysis at both temperatures. The peak of the tumor necrosis factor-&agr; mRNA expression level was observed at 1 hr after the stimulation at 37°C and 2 hrs after the stimulation at 33°C. In the interleukin-1&bgr; mRNA expression, at 37°C the first peak appeared 1 hr and the second 6 hrs after the stimulation. In contrast, at 33°C, the first peak appeared 2 hrs and the second 12 hrs after the stimulation. Whereas interleukin-6 mRNA expression at 37°C peaked 6 hrs after the stimulation, no definite peak was observed at 33°C and the expression level was approximately half of that at 37°C. The maximum intensity of nuclear factor-&kgr;B activation at 33°C was delayed by 1.5 hrs compared with that at 37°C. Conclusions Moderate hypothermia delays the induction of proinflammatory cytokines in human peripheral blood mononuclear cells.

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. DNA-DNA hybridization demonstrated two new similarity groups. From the results of the present study, the names Streptococcus peroris sp. nov. and Streptococcus infantis sp. nov. are proposed for these new groups. The type strains are O-66T (= GTC 848T = JCM 10158T) and O-122T (= GTC 849T = JCM 10157T), respectively.

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

ABSTRACT 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. This study may contribute to the understanding of the pathogenic role of SPE B/SCP in streptococcal infection and streptococcal toxic shock syndrome.

Inhibition of poliovirus by effect of a methylthiopyrimidine derivative.

Summary Ethyl-2-methylthio-4-methyl-5-pyrimidinecarboxylate (S-7) inhibited the cytopathic effect and multiplication of poliovirus in primary monkey kidney cultures but not that of ECHO 1, 5, 13, 22, 23, Coxsackie A7, A9, B3, or B5. It did not inhibit cell multiplication under these conditions. Passage of poliovirus type 2 in the presence of S-7 gave rise to S-7-resistant and S-7-dependent strains. There was no cross resistance between S-7 and guanidine or HBB.

Characterization of recombinant Streptococcus mitis‐derived human platelet aggregation factor

Ohkuni H, Nagamune H, Ozaki N, Tabata A, Todome Y, Watanabe Y, Takahashi H, Ohkura K, Kourai H, Ohtsuka H, Fischetti VA, Zabriskie JB. Characterization of recombinant Streptococcus mitis‐derived human platelet aggregation factor. APMIS 2012; 120: 56–71.

The presence of tumor necrosis factor-α 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-α (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. TNF was found in the sera of four patients (20%) at levels ranging from 10.4 to 53.1 pg/ml, while a positive reaction for the anti-TNF antibody was detected in the sera from six patients (30%), two of whom showed both TNF and its antibody. Thus, either TNF or the anti-TNF antibody was present in the sera from 8 of 21 patients (40%). The results of this study indicate that TNF may be present in the circulation of at least 40% of cachexic patients, and suggest that it may be one of the main mediators of cancer-associated cachexia.

Correlation between epidermal growth factor receptor concentration and the growth of human gastric cancer xenografts in nude mice

SummarySeven human gastric cancer xenografts with different concentrations of EGF receptor were established in nude mice. The expression of EGF receptor in the tumors was demonstrated by Western blotting with anti-EGF receptor antibody, binding assay with125I-EGF and immunohistochemistry with anti-EGF receptor antibody. Western blotting revealed EGF receptor doublet bands at molecular masses of 150 KDa and 170 KDa in all of the samples. The concentration of125I-EGF binding activity in the tumors ranged from 36.0 to 11,000 fmol/mg protein, with a mean of 345 fmol/mg protein. EGF receptor was also demonstrated immunohistochemically on the apical border of the glands and the cell membrane of the tumor cells. There seemed to be a close correlation between the concentration of125I-EGF binding activity and the doubling time of these tumors in nude mice (γ= -0.68). However, no definite correlation was observed between EGF ligand binding and histological features of intestinal type or diffuse type. The expression of EGF receptor appears to facilitate the growth of human gastric cancer xenografts in nude mice.

淋菌外膜蛋白1に対するモノクローナル抗体を用いた淋菌迅速同定法 (GonoGen II) の有用性に関する検討

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. Other Neisseria. spp, H. influenzae, H. parainfluenzae, E. corrodens, M. catarrhalis, and A. baumannii showed negative test results. Non-Neisseriae. spp, such as S. aureus. P. aeruginosa, E. faecalis, and E. coli also showed negative test results. No cross-reactivity was found between GC and other Neisseriae. spp or non-Neisseriae. spp. In a mixed suspension of GC and all of non-Neisseriae. spp as mentioned above, the GonoGen II test was positive. The specificity and sensitivity of the test for the identification of GC were 98% and 100%. The minimum limit of detection of GC was > or = 1 x 10(5) cfu/mL. Decision making based on the test result is possible within 10 minutes. These findings also suggest that the test does not require pure GC. The GonoGen II test appears to be a reliable, quick and easy-to-use assay, and also to not require viable GC. Thus GonoGen II is shown to be a very useful test for the identification of GC.

[HA-and HI-tests for myxoviruses with formalinized erythrocytes].

With formalinized erythrocytes (FRC) from chickens, men and guinea-pigs, hemagglutination and hemagglutination-inhibition test for myxoviruses with Salk pattern method were studied and the results obtained are as follows.1) Spontaneous agglutination of FRC was prevented with 0.05 per cent bovine serum albumin in the suspending medium.2) There was no or little if any differences between HA- or HI-titers obtained with FRC and those with native red cells.3) FRC was available for HI test after the lyophylization.4) Hemadsorption test in HVJ-infected HeLa cells was successfully performed with FRC without substitution of bovine serum albumin.5) HVJ-adsorpting capacity of FRC was identical with that of native cells, but the eluting capacity was decreased in FRC.6) The ABO-antigenicity of human red cells was partially destroyed by the formalinization.