Vol 17, No 2 (2017)

Techniques of mass spectrometric analyses are briefly considered, its fields of application in medical research and clinical diagnostics are reviewed. The determination of biomarkers is discussed. The fields of diagnostics in which mass spectrometry plays an important role, namely, human breath analysis, newborn screening, identification of microorganisms, endocrinology, drug therapy, peptide and protein markers, and mass spectrometric visualization, are characterized.

The aims of the research were to synthesize gene, coding previously designed artificial protein antorex - the antagonist of orexins receptor OxR1, to explore it's expression in Escherichia coli cells and to develop the process of this protein purification from Escherichia coli cell lysates. Methods of chemical and enzymatic DNA synthesis, genetic engineering, selection and chromatography were used. The gene coding antorex protein was synthesized, the plasmid DNA рЕТ-antorex, containing this gene and Escherichia coli strain BL21(pЕT-аntorex) - the stable producer of recombinant protein antorex were obtained. Methods of Escherichia coli producer strain cultivation and antorex protein purification with the use of reverse-phase and ion-exchange chromatography was developed. Large-scale cultivation of producer strain was performed in 10L fermenter and the pilot lot of 480 mg of antorex protein with the purity of 97,5% was produced. The developed methods lead to production and purification of antorex artificial protein for conducting pre-clinical and clinical trials.

The aim of the study was to clarify clinical and morphological criteria for prediction of treatment outcomes gastroenteropancreatic neuroendocrine tumors. Тreatment results were analyzed in 190 patients with neuroendocrine tumors of the pancreas diagnosed in 84 (44,2%), various gastrointestinal tract - 89 (46,3%), liver metastases revealed no localization of primary tumor in 17 (9.5%) of patients. It was determined that the main adverse prognostic factors of the disease are: patient age older than 60 years, tumor localization in the colon and pancreas, the index of proliferative activity (Ki-67) tumor >10%, III-IV stage oncologic process. It is shown that significant prognostic factor in patients with generalized forms of the disease is the possibility of complex treatment allowed to increase the median survival of patients with neuroendocrine tumors 2 times and neuroendocrine carcinomas 2.7 times. The findings suggest that the presence of established clinical and morphological criteria of adverse prognosis of neuroendocrine tumors is advisable a more aggressive treatment option to improve long-term results of treatment and quality of life of patients.

One of typical features of neurodegeneration is intranuclear accumulation of proteins containing polyglutamine sequence in their polypeptide chain. The aim of the present study was to identify such proteins, and to determine their distribution in the neurons of the red nucleus in humans not suffered from neurodegenerative diseases. Fragments of the human midbrain were used in the study (n=7). Polyglutamine sequence (poly-Q) in proteins was revealed using immunocytochemistry to monoclonal mouse antibody (clone 1C2). Analysis of the preparations revealed the presence of a weak diffuse reaction in the cell nucleus structures in most cells (both nerve and glial) of the red nucleus. In large neurons in two of the seven cases studied, the reaction of the intranuclear structures was diffuse, and in the other five cases, slightly contoured 1C2-immunopositive accumulations, always located outside of the nucleolus, were determined in the nucleus of large neurons. The obtained data suggest that the polyglutamine proteins typical for pathological structures can be accumulated in nucleus of nerve and glial cells of the red nucleus in humans without neurodegenerative pathology.

The aim was to clarify the phenomenology of electrophysiological pattern of blink reflex. Blink reflex was recorded by electromyography "Neuro-MEP" firm "Neurosoft": the standard procedure was stimulated by the first branch of the trigeminal nerve electrical pulse duration of 100 microseconds amperage of 30-80 mA. The functional test with opening and closing of eyes was used during registration. The slow drift of contours during the registration blink reflex represents one side of the bioactivity of the circular muscles of the eye. The variability of the amplitude of the long-wavelength component is a functional viability suprasegmental modulation systems, extrapyramidal first. It is advisable to study the variability of the long-wavelength component’s characteristics in cases of the functional activity’s disorganization of the afferent, vegetative and extrapyramidal cerebral systems, and the muscular receptor apparatus.

Research objective: The aim of this work was a comparative study of the contents of the Chlamydia pneumoniae and TNF-α in the unstable and stable atherosclerotic lesions in humans and their role in the destabilization of atherosclerotic plaques. Material and methods: Investigated 50 aortic tissue samples received from 61 ± 7 years men who died from coronary heart disease. Verification of unstable progressive atherosclerotic plaques was conducted using histological staining Oil Red O and hematoxylin Mayer. Chlamydia pneumoniae identified in two ways: by using monoclonal antibodies and by modified Romanowski-Giemsa. Determination of TNF-α was made highly sensitive two-stage biotin-streptavidin method. Analysis of the results was carried out using a light microscope Leica DM 2500 and computer program «Leica Application Suite Version 3.4.0». Results: In unstable atherosclerotic lesions in the same areas and cell types that make up the inflammatory infiltrates, as well as in cells smooth muscle cells of these plaques are detected simultaneously Chlamydia pneumoniae and powerful inflammation cytokine TNF-α. In stable lesions intra- and extracellular expression of this cytokine is not detected. Conclusion: On the formation of unstable plaques a large impact could have the combined effect of Chlamydia pneumoniae and TNF-α, contributing to the development of the immunoinflamatory process in the vascular wall. Potential mechanism for strengthening action on the severity of the disease may be associated with direct or indirect influence of infection, as damage to the vascular wall, and increased production of proinflammatory cytokines cells, which are involved in the destabilization of plaque.

To study clinical and epidemiological aspects of prevalence risk factors of cardiovascular diseases among the population of the Novgorod Region. It is 1% one-time sample survey which allows to obtain reliable estimates of health numerical parameters (eg, true incidence, the specific characteristics of identified diseases) among the population of a region, which acts as the total population, while studyiing only a part (sample). The results of the sample study were extrapolated to the general population to obtain the minimum and maximum values. Odds Ratio method was used as evidence-based healthcare method. During mathematical analysis the following methods were used: descriptive statistics; univariate and multivariate dispersion analysis; correlation and regression analysis based on the matrix method and multivariate analysis with a selection of regression equation model (in this case) and evaluation of the results based on a Gaussian distribution; multivariate statistical methods, including cluster and factor analysis. Statistical data processing was conducted using STATISTICA 10, Stat Soft Programme, License number AGAR207F394525FA-6. Among all risk factors, statistically significant differences were found by a dieting factor when t=7.27 (р<0.0001) and t=-2.82 (р<0.0156) in cases of non-dieting assessment. According to nonparametric statistics, interquartile amplitude value (ICD) when following the diet was Me=28.8 (at 26.3-30.8) and Me=70.7 (at 60.7-71.3) in cases of not following the diet. It was found that dieting is more prevalent in women at around 30% range. Within 60-70% of the respondents regardless of gender do not follow the diet. This fact is further reflected in the frequency of dyslipidemia cases in the population. Parametric analysis revealed significant differences in the frequency of smoking, taking into account gender differences. So, 62.4% of respondents never smoked at t=4.91 (р<0.0004). Non-parametric analysis showed that 59% of respondents have never smoked with ICD value as=52.6-69.8. At the same time the maximum values for this indicator were more than 80%. Attention is drawn to the high Gaussian density, equal to 17.1. When evaluating vodka consumption at 50 -100 ml amount, the differences significance was at t=-3.56 (р<0.004). Non-parametric analysis showed that 50 -100 ml of vodka is consumed by 12% of the population with the largest ICD=7,4-12,9, and the Gaussian density value was only 5.5. The highest significance of gender differences is observed in the evaluation of work motivation. In this case, 13.3% of females state the lack of work motivation or moderate work motivation 34.5% (M=13.3, with SD=4.3±3.7 and M=34.5 with SD=15.0±12.1 respectively). Among men, this figure is at M=12.4 SD=2,9±1,9 and M=31.5 with SD=15.2±13.2, respectively. Non-parametric data on lack of motivation on a median value were 11, 1 with ICD=9.3-16.9 among females; Me=12.7 with ICD=11.5-15.0 among males and largest Gaussian density of 3.5, respectively. In the second parameter, according to nonparametric data estimation, there is a fair motivation on the median: 31.7 with ICD=18,6-51,4 among females (Gaussian density was 32.8) , and Me=33.7 with ICD=14.6-47.4 (Gaussian density 32,7) among males, respectively. Thus, it was found that the lack of work motivation in males was more pronounced than in females. When assessing moderate dissatisfaction with family relations, a gender difference in t=2.64 (р<0.0217) was revealed. Among females, M=24.6% with SD=4.5±3.3; among males M=18.4% with SD=4.1±3.1. Non-parametric analysis showed that dissatisfaction with family relations among females was Me=25.0% at ICD=20.9-26.6 (Gaussian density=5.7) and Me=19.5% at ICD=14.4-20.3 (Gaussian density=6.3) among males. Thus, higher levels of moderate dissatisfaction with family relationship are identified among women. For other indicators, there are no statistically significant gender differences are absent. The analysis, based on non-fitting criterion, proves possible impact of such risk factors as obesity, smoking, alcohol abuse (these risk factors are seen more often among younger population), marital status (especially widowers and widows), job dissatisfaction and stress, on the development of cardiovascular diseases in the population. However, high level of educational qualifications in the population allows having special courses for hypertensive individuals where technological intervention acts as a basic method of primary prevention of hypertension and its complications.