Application of bacteriophages in simultaneously controlling Escherichia coli O157:H7 and extended-spectrum beta-lactamase producing Escherichia coli

Applied Microbiology and Biotechnology - Shiga toxin-producing Escherichia coli (STEC) O157:H7 and extended-spectrum beta-lactamase (ESBL) producing E. coli (ESBLEC) are important bacteria of...     

Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology

We previously reported a method to expand human monocytes through lentivirus-mediated introduction of cMYC and BMI1, and we named the monocyte-derived proliferating cells, CD14-ML. CD14-ML differentiated into functional DC (CD14-ML-DC) upon addition of IL-4, resulting in the generation of a large number of DC. One drawback of this method was the extensive donor-dependent variation in proliferation efficiency. In the current study, we found that introduction of BCL2 or LYL1 along with cMYC and BMI1 was beneficial. Using the improved method, we obtained CD14-ML from all samples, regardless of whether the donors were healthy individuals or cancer patients. In vitro stimulation of peripheral blood T cells with CD14-ML-DC that were loaded with cancer antigen-derived peptides led to the establishment of CD4⁺ and CD8⁺ T cell lines that recognized the peptides. Since CD14-ML was propagated for more than 1 month, we could readily conduct genetic modification experiments. To generate CD14-ML-DC that expressed antigenic proteins, we introduced lentiviral antigen-expression vectors and subjected the cells to 2 weeks of culture for drug-selection and expansion. The resulting antigen-expressing CD14-ML-DC successfully induced CD8⁺ T cell lines that were reactive to CMVpp65 or MART1/MelanA, suggesting an application in vaccination therapy. Thus, this improved method enables the generation of a sufficient number of DC for vaccination therapy from a small amount of peripheral blood from cancer patients. Information on T cell epitopes is not necessary in vaccination with cancer antigen-expressing CD14-ML-DC; therefore, all patients, irrespective of HLA type, will benefit from anti-cancer therapy based on this technology.     

Tumor inhibitors having potential for interaction with mercapto enzymes and/or coenzymes: A review

On the basis of extensive information on in vivo metabolism as well as biomimetic reactions using simple SH compounds and some enzymes, numerous chemical functions which react with SH groups are divided into two classes; i.e., one which involves electrophilic addition (EA) to an SH group and the other which features displacement reactions (DR) by the SH group (see Tables 1 and 2). The known tumor inhibitors are accordingly classified into EA and DR types. Biomimetic reactions of tumor inhibitors with model compounds of SH enzymes (or coenzymes) and with some SH enzymes themselves are described. Finally, as enhancement factors for the antitumor activity, the roles of hydrogen-bonding, neighboring group participation, and effect of ester side chains are introduced. These discussions may serve in the development of the new SH alkylating antitumor agents.     

Prolonged pseudoaldosteronism induced by glycyrrhizin.

We describe the natural recovery from the aggravated hypertension, hypokalemia and suppression of the renin-aldosterone axis after the glycyrrhizin discontinuation in two mild hypertensive women aged 71 and 68 years, who had been administered 273 to 546 mg glycyrrhizin daily for 1.5 and 6 months, respectively, for the treatment of liver disease. About one month after the glycyrrhizin discontinuation, acceleration of hypertension, hypokalemia and suppression of the renin-aldosterone system still continued in both patients. At this stage, sodium restriction resulted in the normalization of blood pressure with weight loss and the subsequent sodium repletion produced a rapid increase in blood pressure to hypertensive levels observed before sodium restriction, with weight gain. Plasma renin activity and plasma aldosterone were low and did not respond to sodium restriction. Inappropriately excessive amounts of potassium were also excreted in the presence of hypokalemia. About one and a half months later, the improvements of aggravated hypertension, hypokalemia and suppressed renin-aldosterone system gradually occurred in both patients. Sodium restriction performed about three months later in case 2 no longer produced the changes in blood pressure and body weight. Plasma renin activity and plasma aldosterone responded subnormally to sodium restriction. These results demonstrate that both patients had a prolongation of the syndrome resembling primary aldosteronism except the low plasma aldosterone level about one month after the glycyrrhizin discontinuation. The possible mechanisms by which this prolongation was caused are discussed.     

31P-MRS study of change in intracellular pH during sustained static contractions in human

31P-MRS spectra were obtained from human first dorsal interosseous muscle during and after the voluntary static abduction of the index finger. Endurance tasks were performed at randomly assigned contraction levels of 15, 20, 30 and 40% of maximal voluntary contraction (MVC). Muscle pH was calculated according to Taylor et al. (1983) using chemical shift between inorganic phosphate (Pi) and phosphocreatine (PCr) on the 31P-MRS spectra. Mean values of endurance times of static contractions were 7.25, 5.33 and 3.08 minutes for 20, 30 and 40% MVC, respectively. At 15% MVC, all of the four subjects maintained contraction for 30 minutes, and the contractions were terminated at 30 minutes. Muscle pH at the onset of contractions were 7.12, 6.98, 7.01 and 7.08 for 15, 20, 30 and 40% MVC, respectively. At the end of contractions when the subject could not maintain the force level, muscle pH were 6.07, 5.97 and 5.94 for 20, 30 and 40% MVC, respectively. There was no significant difference in muscle pH at the end of contractions between three conditions by one-way ANOVA. In conclusion, there was a critical muscle pH of about 6.0 where static contractions could not be maintained.