Epidemiological aspects of morbidity in the class of digestive organs in Kazakhstan.

AIM: To study epidemiological aspects of disease, class of diseases of the digestive system in the region of the relative risk in Kazakhstan. MATERIALS AND METHODS: Studied epidemiological parameters are first identified in the incidence and prevalence of class XI ICD in regions of Kazakhstan with the identification of risk relative to the average national level. In a dysfunctional region Kyzylorda region analyzed the performance of its regions and compared with data from medical examination of the population and the survey. RESULTS: A significant excess of the average national incidence rate between 1990 and 2015, was noted for 3 of the 14 regions of Kazakhstan (West-Kazakhstan, Kyzylorda, Mangystau). According to the dynamics of the unfavorable situation on the high level of incidence currently is in the Kyzylorda region (COA). From the areas of the COA maximum, the incidence of diseases of the digestive system registered in Kazalinsk district which exceeded the level at RK 4.8 times. The data on examination and survey of the complaints of the population exceeded the epidemiological indicators. In the area identified as the leading neoplasms of the digestive system in the structure of cancer incidence and exceeded the national average on the related class of blood diseases. Unfavorable situation on the incidence of the digestive system in Kyzylorda region of Kazakhstan is associated with the cancer and blood diseases, this is partly due to the presence of pollutants in the environment.

[Qualitative index of population health losses in the aral sea region].

The article is devoted to the approbation of the modification of the method of the presentation of indices of medical and social losses of the population on an example of environmentally unfavorable Aral Sea region in Kazakhstan. The first step in improvement of the system ofpublic health care is the identification of the most important problems related with the assessment of the disease burden and the adjustment of data of such type in accordance national aims on the decline of medical and social losses in this problem area. So far according to the criteria for the determination of the boundary of ecological catastrophe area one of the indices is the steady gain in the mortality rate. Using the index "Years of Life Lost" (YLL) in medical statistics as a relatively new approach that continues to be improved and its development supposes the improvement in the quality of health statistics. We modified this method consisting in the determination of the coefficient of loss of years. In the given work after comprehensive analysis of the loss of years due to morbidity and mortality on the base of life tables the potential of lost years for the population was determined with bearing in mind of module tables and compared with the loss in years due to the health loss. So far there was declined the dimension of the comprehensive index, there was elevated its comparability with other indices, and the data interpretation has been optimized. Suggested modification can be used for a comprehensive assessment of health state in the execution of epidemiological studies and monitoring of medical and social situation in ecologically unfavorable regions.

[Health state of Aral region dwellers].

Studies of birth rate, natural increment and death rate dynamics of Aral region population helped to set major tendencies in demographic processes for dwellers in ecologic disaster area. Morbidity parameters were evaluated via ICD-10 classes with distribution in age and sex, that helped to identify leading classes of diseases in the population of Kyzylorda region. DALY index parameters were estimated through complex evaluation of health losses due to disablement and mortality among adult population of Kyzylorda region. The studies covered 5 districts of Kyzylorda region (Aral, Kazalin, Zhalagash, Karmaksh, Shiely). The parameters were analysed retrospectively over 10 years--from 2004 to 2013.

[Investigation of the kinetics of insulin amyloid fibrils formation].

Investigation of the structure of ordered protein aggregates--amyloid fibrils, the influence of the native structure of the protein and the environment on the process of fibrillation is currently the subject of intensive research. The present work is devoted to the study of the kinetics of insulin amyloid fibrils formation at low pH values (which are produced at many stages of the isolation and purification of the protein) using a fluorescent probe thioflavin T (ThT). It has been shown that the increase of fluorescence intensity of ThT during the formation of amyloid fibrils is described by a sigmoidal curve, in which 3 areas can be distinguished: the lag phase, the growth and the plateau, which characterize the various stages of fibril formation. Despite the variation in the length of the lag phase at the same experimental conditions (pH and temperature), we have found its reduction with stirring the solution and seeding. Data obtained using electron microscopy showed that the formed fibrils are long, linear filament having a diameter of -20 nm. With increasing incubation time fibril diameter did not change while their length increases to 2-3 µm, which was accompanied by a significant increase in the number of aggregates of fibrils. All the experimental data shows that, regardless of the kinetics of the formation of amyloid fibrils, their properties after the fibrillation process are identical. The results of this work together with the previously studies of insulin amyloid fibrils might be important for clarification the mechanism of their formation, as well as for the treatment of amyloidosis associated with the aggregation of insulin.

[Verification of a decrease in the rigidity of the phage lambda DNA polymeric chain in low ionic strength aqueous solutions by testing the polymer-polymer interlink interactions].

Changes in the rigidity of the polymetric chain of phage lambda double-strand DNA have been studied by laser correlation spectroscopy. It was shown that, as the ionic strength increases, the effect of the screening of the hydrodynamic interaction of the links of the polymeric chain specific for polymeric coils arises in a DNA solution. It is assumed that the screening occurs when the threshold of the overlapping of DNA coils is achieved. The overlapping of coils is the result of a previously observed significant rise of DNA coil size from abnormally small DNA coils in low ionic strength buffers (about 10(-2) M Na+ or less) to maximum possible large coils in the 5SSC and 5SSC-like buffers. Further analysis of the far interlink interactions in linear lambda phage DNA coils in similar buffers at pH 7 and 4 confirms the earlier proposal about the role of H+ ions in the appearance of abnormally small DNA coils. The abnormal decrease in the DNA coil size in low ionic strength buffers is not a specific feature of lambda phage DNA only.

Detection of inflammatory processes during various diseases by the method of flow cytofluorometry.

Oxidative (respiratory) burst is an important manifestation of inflammation. Precise quantitative assessment of this reaction by flow cytometry made it possible to record and evaluate the severity of the inflammatory processes in a wide spectrum of diseases including diphtheria, hepatitis, pneumonia, bronchial asthma, arthritis, vasculitis, postoperative complications, tuberculosis, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, and myocardial infarction. This approach can be employed as a highly sensitive method of detection of inflammatory reactions and monitoring of their course in various pathological processes.

[A cross drug resistance of mammalian cells with a high efficiency of "DNA clearing"]. / Perekrestnaia lekarstvennaia ustoichivost' kletok s povyshennoi éffectivnost'iu protsessa "ochistki DNK".

The process of active dissociation of noncovalently bound agents from DNA or "DNA clearing" in the living cells was described elsewhere. The vital fluorescent bisbenzimidazole dye Hoechst 33342 (4342), which binds tightly but not covalently to DNA in the minor groove, was used for studying interactions of agents noncovalently binding with DNA. The "DNA clearing" is an energy-dependent process, which is suppressed by topoisomerase-II inhibitors and DNA breaks. It has been shown that the rodent fibroblast cell line AA8HoeR-7 is selected for resistance to H342, and characterized by an enhanced dissociation of the bisbenzimidazole dye-DNA complex. This cell line obtained cross-resistance to other DNA damaging drugs: mitomycin C, etoposide and ethidium bromide. That proves that AA8HoeR-7 is cell line with a new mechanism of multidrug resistance.

[Evaluation criterion for nuclear chromatin state of human spermatozoa using flow cytometry--a fertility test]. / Kriterii otsenki sostoianiia khromatina iader spermatozoidov cheloveka s pomoshch'iu tekhniki protochnoi tsitometrii--test na fertil'nost'.

On the basis of the previously described method for estimating the state of the human spermatozoan nuclear chromatin by flow cytometry, we developed a modified system of calculation and presentation of our data. We propose a criterion that allows to classify samples of seminal material according to their fertility potential that could be used for male sterility diagnostics and as a prognostic test for optimization of reproduction programs. The most important theoretical and methodical aspects of the suggested approach are discussed, and comparative analysis of suggested approach and routine flow cytometry SCSA method is made.

Relationship between abnormal sperm chromatin packing and IVF results.

This study was initiated to determine the relationship between the fertilizing potential of spermatozoa and abnormalities in the compact packing of their chromatin which occurs in the final stage of male germ cell differentiation. Chromatin packing involves disulphide bridge covalent cross-linking. The degree of packing was determined from the accessibility of DNA to a fluorescent dye, ethidium bromide, following detergent treatment of the spermatozoa. The amount of dye bound was determined by flow cytometry in the presence or absence of heparin, a polyanion which removes only non-disulphide bridge-linked proteins. The results of a number of different sperm samples were compared with their results following in-vitro fertilization, and a relationship between disordered sperm chromatin packing and rates of embryo cleavage was observed. This study suggests that abnormal chromatin packing in spermatozoa may contribute to male fertility.

Active dissociation of the fluorescent dye Hoechst 33342 from DNA in a living cell: who could do it?

It is assumed that DNA in mammalian cells is a dynamic conformationally unstable system. This instability provides the cell with a mechanism for dissociating a large number of substances that bind tightly but not covalently to DNA. Among these is the fluorescent dye Hoechst 33342, which binds to DNA in the minor groove. We have selected cell lines with a high capability for active dissociation of Hoechst 33342. Comparative protein analysis of these lines by means of two-dimensional (2-D) electrophoresis was performed. Cell and nuclear proteins were analyzed from these and normal strains. A few proteins with significantly changed quantities have been found. The preliminary search of the 2-D database allowed us to identity some known and unknown cellular proteins that could participate in active dissociation of the dye from DNA.

[Process of active dissociation of vital stain Hoechst 33342 from DNa in cells of cultured rodent cell lines]. / Protsess aktivnoi dissotsiatsii prizhiznennogo krasitelia Khëkhsta 33342 iz DNK kletok perevivaemykh kletochnykh linii gryzunov.

The process of active dissociation or "DNA clearing" of not covalently binding agents from DNA in living HeLa cells was shown by the flow cytometry technique. The vital fluorescent bisbenzimidazole dye Hoechst 33342, which binds tightly but not covalently to DNA in a minor groove, was used as a basic model to study the interaction of not covalently binding agents with DNA. In this paper, we continue to analyse the "DNA clearing" process in the living fibroblasts of different rodent species (mouse, rat, Chinese hamster). The obtained data suggest that the processes of active dissociation or "DNA clearing" of Hoechst 33342 have some common features in all investigated mammalian cells. Nevertheless, some differences in the process were found in these lines. The role of nucleotide excision repair genes in the process of DNA clearing was not established.

[The production of clones of "man x Chinese hamster" hybrid cells containing different parts of the human genome]. / Poluchenie klonov gibridnykh kletok "chelovek x kitaiskii khomiachok", soderzhashchikh raznye chasti genoma cheloveka.

Some approach has been described to create hybrid cell lines (human x Chinese hamster) which contain different parts of human genome, and then efficiently to reveal and isolate the human DNA from these. This method involves the introduction of a selective marker in different sites of the human cell genome, by transfecting them with plasmid SV2neo, and the use of flow cytometry and DNA polymerase chain reaction with primers specific only for human DNA.

[Cell-by-cell flow cytometric analysis of chromosomes]. / Pokletochnyi protochnotsitometricheskii analiz khromosom.

The authors discuss various aspects of a recently developed method permitting a detailed flow cytometric analysis of the individual cell karyotypes such as instrumentation, histochemistry, data proceeding algorithms. Possible drawbacks of the method and the ways of their overcoming are considered. Results of analysis of the Chinese hamster cells are presented that illustrate the possibilities of the method, including the metaphase chromosome distribution according to their fluorescence intensity, the analysed cell distribution according to their chromosomes number, the table in which the individual cell karyotypes are distributed according to their fluorescence. The results obtained show that the developed method may be successfully used for investigating chromosomal iNstability and heterogeneity of the mammalian cells.