Epidemiological problems of ozena and the means for controlling this infection

Clinical, bacteriological, serological and epidemiological studies of ozena morbidity among the population of Minsk were carried out in 1970-1980. On January 1, 1981, the ozena morbidity rate among the inhabitants of Minsk was 26.72%. Ozena was found to affect mainly children and women. A wide spread of the family foci of this disease (31.68%) was revealed. The results of this study indicate that the source of K. ozaenae is a sick person who begins to excrete the bacteria in the prodromal period of the disease and may continue to excrete them for many years. The transfer of K. ozaenae occurs probably by droplet or contact infection. The droplet infection is less active in the absence of symptoms (coughing, sneezing) facilitating excretion of the infective agent into the air and in cases of the low susceptibility of persons to ozena. The main measures for controlling ozena are the timely detection and sanitation of the sources of ozena, as well as the current disinfection of the infection foci in apartments.     

Viability and proliferative activity of a Chinese hamster cell culture exposed to nitrosoureas in exponential and stationary growth phases

The lethal effect of antitumor nitrosourea chloroethyl derivatives on proliferating (exponential phase of growth) and non-proliferating (stationary phase of growth) cells is observed at a concentration 5-fold less than that of methyl derivatives revealed by the colony-formation technique. 1,3-bis(2-chlororoethyl)-1-nitrosourea is equally effective towards proliferating and non-proliferating cells, but chlorozotocin exerts a primary cytotoxic effect on proliferating cells. 1-methyl-1-nitrosourea at low concentration causes death more readily of proliferating cells than non-proliferating ones. However, studies on proliferative activity during the first hours after treatment with 1-methyl-1-nitrosourea revealed drug sensitivity in cells being at the early stationary phase of growth.     

Relation between the changes in the structure and synthesis of DNA in the cells of mouse leukemia L1210 induced by 1-methyl-1-nitrosourea and 1,3-bis(2-chloroethyl)-1-nitrosourea

The damage of DNA structure and synthesis in murine leukemia L1210 cells upon single administration in therapeutic doses of antitumour agents of N-nitrosourea type, such as 1-methyl-1-nitrosourea (MNU) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) was studied. MNU and BCNU were characterized by stronger inhibitory effects on de novo DNA synthesis compared to additional pathway of DNA synthesis in leukemia L1210 cells in vivo. Centrifugation in alkaline sucrose density gradients of L1210 cell lysates has revealed persistent single-strand breaks and alkaline-labile sites in newly replicated DNA. Parental DNA structure was more stable to damaging drug effects than that of newly replicated DNA. The results are consistent with our previous data on the differences in the mechanisms of MNU and BCNU action and the absence of complete cross resistance between the drugs.     

Excitotoxic Insult Results in a Long-Lasting Activation of CaMKIIα and Mitochondrial Damage in Living Hippocampal Neurons

Over-activation of excitatory NMDA receptors and the resulting Ca²⁺ overload is the main cause of neuronal toxicity during stroke. CaMKII becomes misregulated during such events. Biochemical studies show either a dramatic loss of CaMKII activity or its persistent autonomous activation after stroke, with both of these processes being implicated in cell toxicity. To complement the biochemical data, we monitored CaMKII activation in living hippocampal neurons in slice cultures using high spatial/temporal resolution two-photon imaging of the CaMKIIα FRET sensor, Camui. CaMKII activation state was estimated by measuring Camui fluorescence lifetime. Short NMDA insult resulted in Camui activation followed by a redistribution of its protein localization: an increase in spines, a decrease in dendritic shafts, and concentration into numerous clusters in the cell soma. Camui activation was either persistent (> 1–3 hours) or transient (~20 min) and, in general, correlated with its protein redistribution. After longer NMDA insult, however, Camui redistribution persisted longer than its activation, suggesting distinct regulation/phases of these processes. Mutational and pharmacological analysis suggested that persistent Camui activation was due to prolonged Ca²⁺ elevation, with little impact of autonomous states produced by T286 autophosphorylation and/or by C280/M281 oxidation. Cell injury was monitored using expressible mitochondrial marker mito-dsRed. Shortly after Camui activation and clustering, NMDA treatment resulted in mitochondrial swelling, with persistence of the swelling temporarily linked to the persistence of Camui activation. The results suggest that in living neurons excitotoxic insult produces long-lasting Ca²⁺-dependent active state of CaMKII temporarily linked to cell injury. CaMKII function, however, is to be restricted due to strong clustering. The study provides the first characterization of CaMKII activation dynamics in living neurons during excitotoxic insults.     

Biochemical mechanisms of resistance to a new antineoplastic drug CRC 680578 from the nitrosourea class]

The biochemical mechanisms of resistance to CRC 680578, a new antitumour chloroethylnitrosourea alpha-amino acid derivative, were studied. Alterations in DNA, RNA and protein syntheses, SH-group content, drug efflux, activities of replicative and repair enzymes, such as ribonucleotide reductase, thymidine kinase, O6-alkylguanine-DNA-alkyltransferase and DNA polymerases alpha and beta and damages of the DNA secondary structure were investigated in sensitive and resistant to CRC 680578 leukemia L1210 cells. It was found that the total SH-group number in drug-resistant cells was increased (about 1.3-fold in comparison with sensitive cells) which seems to be due to the mechanisms of drug resistance. CHC 680578 induced less pronounced inhibition and more rapid restoration of DNA and RNA synthesis in resistant cells. No differences between the ribonucleotide reductase and thymidine kinase activities were found either in intact cells of the both strains or after drug administration. The efficiency of repair of DNA chloroethyl adducts by O6-alkylguanine-DNA-alkyltransferase in leukemia cells of various sensitivity was found to be identical. The differences in enzyme activities in intact cells of the both strains were insignificant. It was supposed that factors other than changes in the level of O6-alkylguanine-DNA-alkyltransferase in leukemia cells may be responsible for the resistance to CRC 680578. The increase in the levels of DNA polymerase alpha and, especially, of DNA polymerase beta, in sensitive (but not resistant) mouse leukemia cells 48 hours after drug administration is though to define the mechanism of resistance to the new antitumour agent CHC 680578.     

Dynamics of a vortex with the U-shaped filament (ends “anchored”) in the heart of the ground squirrel

The dynamics of an electrical scroll wave with the U-shaped filament with both ends of the filament being “anchored” on the endocardial surface and the dependence of the structure of pseudoECG on the dynamics of the vortex during the development of polymorphic tachysystolia have been studied by applying premature stimuli to the “target phase” with subsequent registration of the spatial and temporal distribution of electrical potential throughout the surface (endocardial and epicardial) of a thin (≈1 mm) preparation. It was found that (1) the pseudoECG of the polymorphic form during the tachysystolia attack can be observed in the case that the position of the filament ends on the surfaces of the preparation does not practically change from turn to turn (filament ends are “anchored”); (2) the thread of a scroll wave during this attack can twist and untwin (twisted filament), just as it was the case for scroll waves with a straight filament; (3) in the case of pseudoECG of polymorphic form, the twisting and untwining of the filament were stronger (the angle of maximal twisting was 120 degrees and more), and the angle of twisting changed by a substantially greater value from turn to turn as compared with the pseudoECG of monomorphic form; (4) in the case of pseudoECG of polymorphic form, the time interval between the appearance of waves on the surfaces of the preparation (T _epi-endo) was substantially greater and changed to a greater extent from turn to turn of the vortex; and (5) simultaneously with the appearance of pseudoECG of polymorphic form and the onset of changes in the twisting of the scroll and the T _epi-endo interval indicated in (2–4), significant changes in the patterns of coverage of the surface by excitation occurred. Based on the results obtained, an explanation of the reasons for the appearance of excitation breakdown patterns on the surface of the myocardium was proposed, which differs from the traditional viewpoint. These patterns may be the result of reflection on myocardial surfaces of the activity of not different simultaneously occurring sources of initiation of excitation but of a single three-dimensional vortex whose filament twists when passing through the thickness of the myocardium and can closely approach one or the other surface.     

Peptides analogous to tethered ligands liberated by activated protein C exert neuroprotective effects in glutamate-induced excitotoxicity

Haemostatic proteinases may appear in brain tissue after injury and under inflammation as a result of the blood-brain barrier disruption. Serine proteinases regulate cell functions through G-protein-coupled transmembrane protease-activated receptors (PARs). Proteinases cleave only one peptide bond of receptor exodomain, which results in the formation of a new N-terminus (“tethered ligand”) that can specifically interact with the second extracellular loop of the receptor and activate it. Two types of receptors (EPCR and PAR1) are necessary for the cytoprotective effect of activated protein C (APC) on endothelial cells and neurons. APC activates PAR-1 and controls gene expression of proinflammatory and proapoptotic factors. APC exerts a protective effect in stressed neurons and hypoxic brain endothelium, modulates the activity of endothelial cell genes involved in apoptosis, and stabilizes the endothelial barrier. We suppose that the peptides analogous to the PAR1 tethered ligand released by APC may have a neuroprotective effect similar to that of APC. We have simulated ischemic brain damage using a model of glutamate excitotoxicity on the primary culture of neonatal rat hippocampal neurons. It was shown that NPNDKYEPF-amide (peptide 9) and NPNDKYEPFWE (peptide 11) more effectively reduced the level of apoptosis during neuronal excitotoxicity in comparison with APC, while the influence of these peptides on the number of living and necrotic cells was analogous to that of APC. The findings suggest that the protective effect of the peptides analogous to the PAR1 tethered ligand is comparable to the protective effect of APC under glutamate excitotoxicity. Investigation of the mechanisms of PAR1 agonist peptides action and development of their shortened versions with high neuroprotective activity may be a relevant approach to the search of novel neuroprotective drugs for treating neurodegenerative diseases and strokes.