Identification of cells with lowered sensitivity to cytostatic agents by the use of fluorescent dyes

With a help of stepwise increase of vincristine concentrations in culture medium several lines of mouse myeloma X63 Ag 8.863 cells resistant to low concentrations of vincristine (6-35-fold) were selected. Rhodamine 123 stained resistant cells and wild-type cells with an equal intensity. However, resistant cells differ significantly from the sensitive ones by the rate of rhodamine efflux. The rate of the efflux was in proportion to the degree of resistance. The efflux of the dye could be blocked by the addition to reserpine, the inhibitor of multidrug resistance. Thus, fluorescent dyes can be used for the detection of cells with low levels of multidrug resistance.     

Results of using an accelerated method of determining microbial flora sensitivity to antibacterial preparations in the surgical clinic]

Express diagnosis using 2,3,5-triphenyltetrasolium chloride as the redox indicator provided in most tests rapid and sufficiently precise determination of the microbial flora sensitivity to antibacterial drugs permitting to start in time the antibiotic therapy of the patients. For rapid response it proved to be useful to incubate beforehand the test material taken from surgical patients within 16 to 18 hours and to increase the indicator concentration up to 2--3 per cent.     

Method of determining the sensitivity of Staphylococcus aureus to methicillin]

The effect of the incubation temperature, i.e. 37 and 30 degrees and the effect of addition of 5 per cent of sodium chloride to the nutrient medium was studied with respect to the results of sensitivity determination of staphylococci to methicillin by 3 methods, such as serial dilutions in liquid and solid media and replica method applied to separate colonies. With the use of all 3 methods incubation of the samples at a temperature of 30 degrees increased the rate of staphylococcal cultures resistant to methicillin. Addition of sodium chloride to the solid nutrient medium increased the level of detecting methicillin resistant cultures in the samples incubated at a temperature of 37 degrees and decreased it in the samples incubated at 30 degrees.     

A method for determining the sensitivity of the causative agents of suppurative infection to iodopiron

Bacterial suspensions of cultures (10(7) PFU) of pathogenic Staphylococcus, vulgar Proteus and pyocyanic bacterium are resistant to 2.9-46.87 mg per disc of iodopyron in 100% of cases. Sensitivity of 32 and 100% of cultures of pyocyanic bacteria (of 25 under study) being in the suspension (10(5) PFU) to 11.71 and 23.43-9.375 mg of idopyron, respectively, in a hole has been established. To determine sensitivity of pyocyanic bacteria to iodopyron it is suggested to use the procedure of adding of 11.71 mg of the preparation (water solution) to the holes in MPA.     

Cytotoxic and cytostatic effect of avermectines on tumor cells in vitro]

Effect of natural avermectin complex (aversectin C) and separate avermectins A1, A2, B1 and B2 in the cell culture of murine myeloma Ns/o, Erlich carcinoma ascites and human larynx carcinoma Hep-2 was investigated. It was shown that aversectin C within the concentrations of 0.1 to 1.0 mcg/ml inhibited proliferation of tumor cells and induced their death. Proliferation inhibition was due to the delay of the cells cycle start (lag-phase prolongation) and blocking of mitotic cycle. Ns/o cells death had apoptosis signs: chromatin condensation and fragmentation, DNA fragmentation. It was demonstrated that only avermectin A1 has cytotoxic activity within the concentrations used, avermectins A2 and B2 had cytostatic activity, avermectin B1 showed no activity under the experimental conditions.     

Elaboration of a rapid method of determining the sensitivity of Brucella to antibiotics]

Optimal conditions were developed for determination of antibiotic sensitivity in Brucella by using enzyme immunoassay directly in the primary cultures of the material tested. The Brucella concentration in the material tested should be not lower than 1.10(6) microbial cells/ml and the time of culture incubation be 24 hours at 37 degrees C. The obligatory condition is to use a liquid medium, i.e. the Albimi broth with 1% glucose. To inhibit the foreign microflora it is recommended to use polymyxin B and amphoglucamine in a concentration of 3 microgram/ml. The use of enzyme immunoassay was shown that it was possible to determine the antibiotic sensitivity of Brucella in practice.     

A cassette micromethod of determining the sensitivity of anaerobic microorganisms to antimicrobial drugs]

A micromethod based on the use of plates with wells and microquantities of microbial suspensions is described. It provides determination of MICs of antibacterial drugs and sensitivity of clinical strains of anaerobes of 2 or 3 species predominating in pathological materials as well as to preliminarily identify some anaerobic bacteria by their antibiotic sensitivity.     

Dynamics of the change in sensitivity of shigellae to gentamycin and cephaloridine in vitro]

The dynamics of the changes in the Shigella sensitivity to gentamicin and cephaloridin was studied in vitro using liquid nutrient media with gradually increasing concentrations of the drugs. 50 passages were performed. It was found that Shigella flexner and sonnei decreased their sensitivity to gentamicin to a little extent and remained middle sensitive. Sensitivity of Shigella flexner to cephaloridin also changed to a little extent, while Shigella sonnei became moderately resistant.     

Antibiotic sensitivity of the causative agents of suppurative surgical infection]

Sensitivity of 1492 strains of the causative agents of the surgical purulent infections, i. e. pathogenic Staphylococcus, Proteus, Ps. aeruginosa and E. coli to benzylpenicillin, streptomycin, levomycetin, tetracycline, erythromycin, oleandomycin, monomycin, kanamycin, novobiocin, ampicillin, oxacillin, ceporin, gentamicin and rifampicin was studied. Gentamicin was most active against all the bacterial species tested. The staphylococci were in addition sensitive in a high percentage of the cases to rifampicin, novobiocin, ceporin, monomycin and kanamycin. The isolates of E. coli were in addition sensitive to ceporin, monomycin and kanamycin. Sensitivity of the strains of Ps. aeruginosa and Proteus was low to all of the antibiotics except gentamycin. Most of the strains of the causative agents of the surgical purulent infections were multiresistant to 4 antibiotics. The number of the staphylococcal strains sensitive to benzylpenicillin, streptomycin and levomycetin increased in 1976 as compared to 1975 on the background of a limited use of these antibiotics in clinics.     

Membrane mechanisms of the effect of alkylating cytostatic agents

The data bearing witness to realization of the cytostatic activity of alkylating agents by means of interaction with cell membrane receptors was obtained. Alkylating agents block receptors of the polyphosphoinositol system. It is shown that chloralkylamines inhibiting sites coincide with M-++cholinomimetics and alpha-adrenergic receptors, inhibitors of H-histamine receptors, cyclo- and lypoxigenase inhibitors. Such likeness is determined by identical structures of the molecule active part, and quantum-chemical calculations.     

Optimal assay design for determining the in vitro sensitivity of ring stage Plasmodium falciparum to artemisinins

Recent reports demonstrate that failure of artemisinin-based antimalarial therapies is associated with an altered response of early blood stage Plasmodium falciparum. This has led to increased interest in the use of pulse assays that mimic clinical drug exposure for analysing artemisinin sensitivity of highly synchronised ring stage parasites. We report a methodology for the reliable execution of drug pulse assays and detail a synchronisation strategy that produces well-defined tightly synchronised ring stage cultures in a convenient time-frame.     

GRP78 regulates sensitivity of human colorectal cancer cells to DNA targeting agents

This study was carried out to investigate the activation status of unfolded protein response (UPR) in colorectal cancer (CRC) and its contribution to CRC resistance to chemotherapy-induced apoptosis. Chemotherapy-induced apoptosis was assessed by the propidium iodide method. Activation of UPR was evaluated in CRC cell lines using immunoblotting technique and in CRC tissues using immunohistochemistry. Findings of the present study revealed that the UPR is constitutively activated in CRC cell lines and CRC tissues isolated from patients, as evidenced by relatively high levels of the 78-kDa glucose-regulated protein (GRP78) and spliced X-box-binding protein 1 mRNA in tissue samples. In addition, CRC cell lines differentially responded to clinically relevant DNA-targeting agents including cisplatin, and 5-flourouracil. Moreover, the levels of GRP78 were inversely associated with sensitivity of CRC cells to chemotherapy-induced apoptosis. Inhibition of GRP78 by siRNA resulted in increased sensitivity of CRC cells to chemotherapeutic agents. Collectively, current results appear to provide novel insights into the role of UPR in determining sensitivity of CRC cells to chemotherapeutic agents and might have important implications for personalized CRC treatment.