Vol 71, No 2 (2016)

BIOCHEMISTRY: CURRENT ISSUES

Influence fatty acids on oxygen consumption by isolated rat cardiomyocytes in ischemic or diabetic heart disease

Afanasiev S.A., Egorova M.V., Kutsykova T.V., Popov S.V.

Abstract

Background: one of the reasons of violation of the functional viability of the myocardium is considered to be the shortage oxygen and energy of the myocardium. Its reason is the inhibitory effect of fatty acids on glucose oxidation. Recently, however, new data have been published, proving the need for and importance of fatty acids in the maintenance and regulation of the functional activity of the myocardium in chronic pathology.

Objective: to investigate the influence of free polyunsaturated and saturated fatty acids (FA) on the oxygen uptake of isolated cardiomyocytes in intact rats and animals with ischemic or diabetic heart disease.

Methods: executed non-randomized controlled study. Considered 3 groups of male rats of Wistar line (weight 250-300 g) with 10 animals in each group. Myocardial infarction ("heart attack") caused by ligation of the left coronary artery, and diabetes ("diabetes") – intraperitoneal injection of streptozotocin. The group "control" was presented intact animals. Myocardial infarction caused by ligation of the left coronary artery, and diabetes by intraperitoneal injection of streptozotocin. Isolated cardiac myocytes were obtained by enzymatic method. Oxygen consumption was assessed polarographically at different saturation incubation medium with oxygen ([O2] £8 mg/l and ([O2] ≥16 mg/l).

Results: As the fatty acid used arachidonic and palmitic acid. It is established that the introduction to the incubation medium 20 μm arachidonic or palmitic fatty acid significantly increased the oxygen consumption of intact cardiomyocytes of rats. Both at the ischemic and at diabetic defeat of heart was obtained the opposite result. The most pronounced decrease of oxygen consumption are shown in diabetes mellitus.

Conclusion: Conclude that the inhibitory effect of LCD on the rate of oxygen consumption during the at the ischemic and at diabetic defeat of heart may be due to their influence on the barrier function of mitochondrial membranes of cardiomyocytes, the activity of membrane-associated enzymes and their associated processes.

Annals of the Russian academy of medical sciences. 2016;71(2):97-101
pages 97-101 views

INTERNAL DISEASES: CURRENT ISSUES

Hepatobiliary system diseases as the predictors of psoriasis progression

Smirnova S.V., Barilo A.A., Smol'nikova M.V.

Abstract

Purpose of the study. To assess the state of the hepatobiliary system in psoriasis and psoriatic arthritis in order to establish a causal relationship and to identify clinical and functional predictors of psoriatic disease progression.

Methods. The study included patients with extensive psoriasis vulgaris (n = 175) from the age of 18 to 66. Divided into 3 groups: 1 – psoriasis patients with isolated skin lesions (n = 77), 2 – patients with psoriatic arthritis (n = 98), 3 – apparently healthy blood donors (n = 30), matched by sex and age. The evaluation of functional state of the hepatobiliary system was performed by analysis of the clinical and anamnestic data and by the laboratory- instrumental methods.

Results. Identified predictors psoriasis: triggers (stress and nutritional factor),  increased total bilirubin, aspartate aminotransferase, alkaline phosphatase, gamma glutamyl transferase, eosinophilia, giardiasis, carriers of hepatitis C virus, and changes focal ductal formation in the liver , thickening of the walls of the gallbladder by ultrasound. Predictors of psoriatic arthritis: age over 50 years, dyspeptic complaints, the presence of hepatobiliary system diseases, the positive right hypochondrium syndrome, the clinical symptoms of chronic cholecystitis, excess body weight, high  levels of bilirubin, cholesterol and low density lipoprotein, hepatomegaly, non-alcoholic fatty liver disease.

Conclusion. High activity of hepatocytes cytolysis, cholestasis, inflammation, metabolic disorders can be considered psoriatic arthritis as a severe clinical stage psoriatic disease where the hepatobiliary system, in turn, is one of the main target organs in systemic psoriatic process. Non-alcoholic fatty liver disease and chronic cholecystitis are predictors of psoriatic disease progression.
Annals of the Russian academy of medical sciences. 2016;71(2):102-108
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MICROBIOLOGY: CURRENT ISSUES

Evaluation of the recombinant protein Tp0965 of Treponema pallidum as candidate antigen for serological diagnosis of syphilis

Runina A.V., Starovoitova A.S., Deryabin D.G., Kubanov A.A.

Abstract

Backgraund: Treponemal tests based on detection of antibodies against the T. pallidum antigens are the most specific methods for serological diagnosis of syphilis. Due to inability to cultivate this bacterium in vitro, the most promising sources of its antigens for diagnostic reactions are recombinant proteins of T. pallidum. Cloning and evaluation of the analytical value of certain T. pallidum proteins is the approach to improve the sensitivity, specificity, and reproducibility of serological tests for syphilis, including possibilities of differential diagnosis of various forms of the disease.

Objective: The aim of present study was to evaluate the analytical values (sensitivity and specificity) of Tp0965 recombinant protein of T. pallidum as a candidate antigen for serological diagnosis of syphilis.

Methods: Tp0965 gene was amplified by polymerase chain reaction from T. pallidum genomic DNA (Nichol's strain) and the nucleotide sequence was cloned into the expression vector pET28a. E. coli BL-21 (DE3) cells were transformed with this plasmid, and the recombinant protein production was induced by isopropyl-β-D-1-thiogalactopyranoside. Isolation and purification of Tp0965 recombinant protein from bacterial lysate was performed by metal‐chelate affinity chromatography using Ni-NTA Sepharose. The collected protein was seeded on high binding 96-well plates, which were used for ELISA with sera of patients with various forms of syphilis in comparison with healthy donors.

Results: High frequency of positive ELISA results was shown with serum of patients with syphilis, compared with a group of healthy donors. The sensitivity of serological reactions using recombinant protein Tr0965 was 98.8%, specificity - 87.5%. The highest sensitivity (100%) was detected in the groups of patients with primary, secondary and early latent syphilis, while in the group of patients with late latent syphilis it decreased to 95.2%.

Conclusions: We concluded that due to their specificity the recombinant protein Tp0965 T.pallidum can be used as a candidate antigen for development of syphilis serological diagnostic assays. 

Annals of the Russian academy of medical sciences. 2016;71(2):109-113
pages 109-113 views

ONCOLOGY: CURRENT ISSUES

The effects of cysplatin on human adipose tissue derived mesenchymal stromal cells under different oxygen levels

Rylova Y.V., Buravkova L.B., Zhivotovky B.D.

Abstract

Objective: To evaluate the damaging effects of cisplatin on MMSCs from adipose tissue in a phase of active proliferation and the state of the monolayer, which was exposed at standard (20%) and reduced to 1% and 5% level of oxygen.

Methods: The incubation MMSC with cisplatin was performed on cultures of 2 passage in a state in monolayer and cultures in the active growth phase. Profile surface markers of MMSC determined by flow cytometry. MMSCs viability after incubation with cisplatin was detected by the number of apoptotic and necrotic cells using ANNEXIN V-FITC - PI Kit (Immunotech, France). Standard culture conditions (~ 20% O2) created in a CO2 incubator (Sanyo, Japan), 5% O2 created using multigas incubator (Sanyo, Japan), 1% O2 - using an airtight chamber (Stemcell Technologies, USA).

Results: Incubation of monolayer MMSC with cisplatin at a concentration of 10 ug/ml for 72 hours leads to death of half of the cells in culture under 20% O2, 5% O2 and 1% O2. Cisplatin increased the fracture of PI+-cell, and PI+/Ann+-cells under all culture conditions. The short-term exposure with cisplatin (24 and 48 hours) did not cause the damaging effect. Effects of cisplatin on the MMSC in the growth phase for 48 hours led to accumulation of Ann+-cells and PI+/Ann +-cells under all culture conditions. However least damaging effect of cisplatin was observed in culture under hypoxic conditions (1% O2).

Conclusion: These data suggest that monolayer MMSCs are dying primarily through necrosis, whereas MMSC in the growth phase in response to cisplatin treatment are dying by apoptosis, regardless the oxygen tension.

Annals of the Russian academy of medical sciences. 2016;71(2):114-120
pages 114-120 views

Melanoma and human papillomaviruses: is there an outlook for study? Review of the literature

Volgareva G.M., Mikhaylova I.N., Golovina D.A.

Abstract

Melanoma is one of the most aggressive human malignant tumors. Its incidence and mortality are growing steadily. Ultraviolet irradiation is the main risk factor for melanoma, however it seems not to be the sole environmental factor involved in melanomagenesis.

Probability of viral etiology of melanoma has been discussed for a long time. Human papillomaviruses have been mentioned among candidates for its etiologic agents. Much interest in these viruses is due to the fact that powerful carcinogens causing cervical cancer and some other cancer types are members of this group. The aim of the review is analysis of literature data on association of melanoma with human papillomaviruses.

Several groups found papillomaviruses in skin melanomas as well as in mucous ones; viruses if high oncogenic risk were detected in some cases. In majority of the studies in which viral DNA-positive melanomas were found testing for viral genome expression was not done. Objective difficulty of such studies is the problem of negative control as papillomaviruses are being routinely found in normal skin and mucous membranes thus creating justifiable threat of tumor specimen contamination with viral DNA in vivo. Complex approach might become helpful in overcoming this obstacle: detecting of viral DNA, viral oncogenes’ mRNAs and corresponding proteins.

There are limited data on aggravation of the disease prognosis in papillomavirus-positive melanomas. However systematic observation of some sizeable patients’ contingents differing in the given tumor character has not been performed yet.

The problem of melanoma association with human papillomaviruses remains unsolved
Annals of the Russian academy of medical sciences. 2016;71(2):121-127
pages 121-127 views

PEDIATRICS: CURRENT ISSUES

Incidence, pathomorphism and outcomes of the bronchopulmonary dysplasia associated with microaspiration of gastric contents

Bryksina E.Y., Bryksin V.S., Pochivalov A.V.

Abstract

Today the influence of the digestive tract functional violations followed by microaspiration of gastric contents (MAGC) on the incidence, features and outcomes of bronchopulmonary dysplasia BPD) remains little studied. Focusing on this aspect makes the research actual.

Objective: determination of the nature of influence of MAGC on the progress and course of BPD.

Methods: 373 newborns exposed to artificial pulmonary ventilation (APV) in the neonatal period were examined. In a tracheobronchial aspirate (TBA) the marker of MAGC – pepsin –was determined. Its activity was measured by extinction value with subsequent analysis of the incidence and nature of the course of bronchopulmonary dysplasia (BPD) in patients against MAGC and without it. During the three years follow-up period outcomes of BPD and features of combined pathology were established.

Results: it was revealed that in children suffered from MAGC the incidence of BPD was higher and grew in proportion to the increase of pepsin activity in TBA and the reduction of gestational age. The extinction increase in TBA values was followed by the increase in duration of APV and the subsequent oxygen therapy, and also the severity of BPD clinical course. By the end of 3 years clinical recovery was detected in 55,2% of children suffering from BPD without MAGC whereas in patients with BPD against MAGC this outcome occured only in 0,9% of cases.

Conclusion: MAGC is clinically significant for the etiology, pathogenesis and pathomorphism of BPD. The proposed method of early detection of MAGC and algorithm of complex therapy can reduce its severity and improve the forecast accuracy of neonatal adaptation. 

Annals of the Russian academy of medical sciences. 2016;71(2):128-140
pages 128-140 views

RHEUMATOLOGY: CURRENT ISSUES

The analysis of efficacy of the use of inhibitor interleukin-1 in the complex therapy of secondary osteoarthritis taking into account the dynamics of clinical and functional indicators

Starodubtseva I.A., Vasilieva L.V., Nikitin A.V.

Abstract

Backgraund: Secondary osteoarthritis (OA) adversely affects an underlying disease and can enhance clinical manifestations of articular syndrome. Early detection of comorbid pathology and early treatment with the therapy of underlying condition is of primary importance to preserve adequate functional activity in patients.

The Aims to analyze the efficacy of the use of inhibitor of interleukin -1 in the complex treatment of secondary OA in rheumatoid arthritis (RA).

Materials and methods: 248 patients with secondary OA in RA were divided in 4 groups: patients of group I (n=62) took inhibitor of interleukin-1 (50mg x 2 times per day) in combination with laser therapy on the basis of methotrexate (10-20 mg per week), group II (n=60) received inhibitor of interleukin-1 in complex with methotrexate, in group III (n=62)-laser therapy with methotrexate and in group 4 – methotrexate. The analysis of treatment efficacy we estimated in 3 and 6 months according to the dynamics of pain on VAS, indexes HAQ and KOOS.

Results: group 1 showed statistical significant reduction of pain on VAS on movement after 3 and 6 months on 28,53 mm (43,6%) and 31,3 мм (48%) accordingly, to values 36,87±1,56* и 33,11±1,11* (p<0,05). It was noted the statistical significant advantages of HAQ dynamics in group I in comparison with groups II, III, IV (p=0,03). The most prominent statistical significant (p=0,02) significant advantages according to the dynamics of index KOOS it was noted in groups of patients who took diacerein (I and II) after 6 months complex treatment.

Conclusions: the involvement of inhibitor of interleukin -1 in the complex treatment of secondary OA in patients with RA contributes to clinical improvement according to VAS (p=0,03), index KOOS, that shows functional condition of the knee and increase the quality of life by index HAQ (p=0,03) after 6 months therapy.

Annals of the Russian academy of medical sciences. 2016;71(2):141-147
pages 141-147 views

Subpopulations of regulatory T-lymphocytes in the peripheral blood of patients with rheumatoid arthritis

Kravchenko P.N., Zhulai G.A., Churov A.V., Oleinik E.K., Oleinik V.M., Barysheva O.Y., Vezikova N.N., Marusenko I.M.

Abstract

Background: Rheumatoid arthritis (RA) is an inflammatory rheumatic disease, associated with a dysfunction of the T cell-mediated tolerance and leading to the disability of working population.  The regulatory CD4+ T cells are play important role in the regulation of autoimmunity and can suppress immune responses. With that, there is no consensus on the content of  these lymphocytes  and their role in the pathogenesis of RA.

Objective: The aim of the study was to assess the content of peripheral blood regulatory T cells (Treg) according to the expression of membrane markers CD4, CD25, CD127 and intracellular FOXP3 marker, as well as the expression of two functional molecules (CTLA-4 and CCR4) in Treg cells of patients with RA.

Methods: Peripheral blood samples of RA patients (mean age 61,1±10,5) and healthy controls (mean age 52,2±14,0) were analyzed. Cell count and the expression level of molecules were assessed by flow cytometry.

Results: Peripheral blood samples of 36 RA patients and 20 healthy donors were analyzed. The number of the cells with Treg-associated phenotypes CD4+CD25hi and CD4+CD25hiCD127low/– was higher in RA patients in comparison with healthy donors. Increased levels of RA CD4+ T cells expressing FOXP3 were also observed. This may be due to increasing in the number of CD4+FOXP3+CD25- lymphocytes, whereas the content of RA CD4+FOXP3+CD25+ Treg cells was at the level of the control. The expression of the functional molecule CTLA-4 in Treg cells of patients with RA was not different from the control, while the expression level of the chemokine receptor CCR4, which provides migration of lymphocytes at sites of inflammation and barrier tissues, was significantly increased in RA patients. 

Conclusion: Increase in the levels of certain Treg-associated lymphocyte populations were detected in peripheral blood of RA patients. During the natural course of RA, alterations in the level of the chemokine receptor CCR4 might indicate the enhanced lymphocyte migration.

Annals of the Russian academy of medical sciences. 2016;71(2):148-153
pages 148-153 views

ENDOCRINOLOGY: CURRENT ISSUES

Obesity: Current Global and Russian Trends

Razina A.O., Runenko S.D., Achkasov E.E.

Abstract

The review of literature presents the results of recent epidemiological studies in obese people in Russia and abroad taking into account gender, age, ethnic, social, and geographical factors. The increase of obesity prevalence among different population groups including children and adolescents was registered. The risks of health problems associated with overweight and obesity probably leading to disability and mortality were analyzed. It was shown that the energy imbalance played a key role in the etiopathogenesis of obesity among many other factors. This occurs as a consequence of discrepancy between energy consumption and energy discharge especially under the conditions of hypokinesia in all spheres of modern life. Particular attention was paid to the analysis of environmental factors, increasing urbanization, and socio-economic conditions of modern life. The fundamental importance of a multidisciplinary approach in the development of prevention and treatment and rehabilitation programs was accentuated. The attention was paid to the role of economic factors in the development of carbohydrate metabolism disorders. The important role of the goverment was shown in the development of health improvement programs including improvement of the environmental situation, change in anthropogenic environment due to physical activity. The preferred direction of comprehensive programs of prevention and treatment of obesity were defined including optimization of the motor regime, diet correction, increasing the motivation to healthy lifestyle, physical education and sports, as well as increasing the individual’s personal responsibility for their health.

Annals of the Russian academy of medical sciences. 2016;71(2):154-159
pages 154-159 views

STATE OF MEDICAL SCIENCES

Technologies for Complex Intelligent Clinical Data Analysis

Baranov A.A., Namazova-Baranova L.S., Smirnov I.V., Devyatkin D.A., Shelmanov A.O., Vishneva E.A., Antonova E.V., Smirnov V.I.

Abstract

The paper presents the system for intelligent analysis of clinical information. Authors describe methods implemented in the system for clinical information retrieval, intelligent diagnostics of chronic diseases, patient’s features importance and for detection of hidden dependencies between features. Results of the experimental evaluation of these methods are also presented.

Background: Healthcare facilities generate a large flow of both structured and unstructured data which contain important information about patients. Test results are usually retained as structured data but some data is retained in the form of natural language texts (medical history, the results of physical examination, and the results of other examinations, such as ultrasound, ECG or X-ray studies). Many tasks arising in clinical practice can be automated applying methods for intelligent analysis of accumulated structured array and unstructured data that leads to improvement of the healthcare quality.

Aims: the creation of the complex system for intelligent data analysis in the multi-disciplinary pediatric center.

Materials and methods: Authors propose methods for information extraction from clinical texts in Russian. The methods are carried out on the basis of deep linguistic analysis. They retrieve terms of diseases, symptoms, areas of the body and drugs. The methods can recognize additional attributes such as «negation» (indicates that the disease is absent), «no patient» (indicates that the disease refers to the patient’s family member, but not to the patient), «severity of illness», «disease course», «body region to which the disease refers». Authors use a set of hand-drawn templates and various techniques based on machine learning to retrieve information using a medical thesaurus. The extracted information is used to solve the problem of automatic diagnosis of chronic diseases. A machine learning method for classification of patients with similar nosology and the method for determining the most informative patients’ features are also proposed.

Results: Authors have processed anonymized health records from the pediatric center to estimate the proposed methods. The results show the applicability of the information extracted from the texts for solving practical problems. The records of patients with allergic, glomerular and rheumatic diseases were used for experimental assessment of the method of automatic diagnostic. Authors have also determined the most appropriate machine learning methods for classification of patients for each group of diseases, as well as the most informative disease signs. It has been found that using additional information extracted from clinical texts, together with structured data helps to improve the quality of diagnosis of chronic diseases. Authors have also obtained pattern combinations of signs of diseases.

Conclusions: The proposed methods have been implemented in the intelligent data processing system for a multidisciplinary pediatric center. The experimental results show the availability of the system to improve the quality of pediatric healthcare. 

Annals of the Russian academy of medical sciences. 2016;71(2):160-171
pages 160-171 views

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