xMAP technology was used for simultaneous identification of six protein toxins (staphylococcal enterotoxins A and B, cholera toxin, ricin, botulinum toxin A, and heat labile toxin of E. coli). Monoclonal antibody-conjugated xMAP microspheres and biotinilated monoclonal antibodies were used to detect the toxins in a sandwich immunoassay format. The detection limits were found to be 0.01 ng/mL for staphylococcal enterotoxin A, cholera toxin, botulinum toxin A, and ricin in model buffer (PBS-BSA) and 0.1 ng/mL for staphylococcal enterotoxin B and LT. In a complex matrix, such as cow milk, the limits of detection for staphylococcal enterotoxins A and B, cholera toxin, botulinum toxin A, and ricin increased 2-to 5-fold, while for LT the detection limit increased 30-fold in comparison with the same analysis in PBS-BSA. In the both PBS-BSA and milk samples, the xMAP test system was 3−200 times (depending on the toxin) more sensitive than ELISA systems with the same pairs of monoclonal antibodies used. The time required for a simultaneous analysis of six toxins using the xMAP system did not exceed the time required for ELISA to analyze one toxin. In the future, the assay may be used in clinical diagnostics and for food and environmental monitoring.
A highly sensitive test system, based on the method of immuno-PCR, was developed for the detection of two staphylococcal toxins: enterotoxin A (SEA) and toxic shock syndrome toxin (TSST). A key element of the developed systems was obtaining supramolecular complexes of bisbiotinylated oligodeoxynucleotides and streptavidin, which were used as DNA tags. Specificity studies showed no cross-reactivity when determining SEA and TSST. The sensitivity of detection of these toxins in the culture supernatants S. aureus was not less than 10 pg/mL.
We studied the effect of combined treatment with cisplatin, glucosaminylmuramyl dipeptide, and TNF-alpha on viability of MCF-7, U-937, B16, and L-929 tumor cells, Ehrlich ascites carcinoma cells, and normal cells (human peripheral blood lymphocytes, peritoneal macrophages, and mouse bone marrow cells). Glucosaminylmuramyl dipeptide was nontoxic for normal and tumor cells, but promoted death of tumor cells after administration in combination with cisplatin and/or TNF-alpha. At the same time, glucosaminylmuramyl dipeptide did not modulate the cytotoxic effect of individual or combined treatment with cisplatin and TNF-alpha on normal cells. Administration of glucosaminylmuramyl dipeptide to cultured MCF-7 cells 20 h before the study increased the potentiating effect of muramyl peptide.
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