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group studied patients with EAA, and distinguishes it from the two comparison groups with acute and subacute stage of the disease.
Taking into account described in the literature familial cases of the susceptibility to the disease, we studied the immunological
changes in 19 patients with a family history on EAA. As can be seen from Table 2 when analyzing immunological parameters in patients of the study group there is an increase of IL-8 to 35,2±4,12 pg/ml by comparison with the group of healthy (18,1±1,81 pg/ml, p<0.02).
Table 2. - The cytokine status in patients with a family burdened EAA
№ Groups of surveyed IL-12 (pg/ml) IL-8 (pg/ml)
1 Practically healthy (n=15) 64,8±8,7 18,1±1,81
2 General group (n=45) 97,9±4,31* 38,4±2,5*
3 Patients with a family burdened EAA (n=19) 60,9±6,8 35,2±4,12*
4 Patients without a family EAA burdeness (n=26) 62,2±5,8 28,1±4,9*
Note: * - p <0.05 - significant differences in comparison with indicators of the group of healthy persons
Levels ofIL-8 in patients EAA with family history constituting production IL-8 and IL-12 have shown that EAA during acute
35.2 ± 4.12 pg/ml exceeded (28,1 ± 4,9 pg/ml) and IL-12 level was course accompanied most pronounced changes in the proinflam-
60.9 ± 6.8 pg/ml had lower values compared with those without matory cytokine IL-8 that provides an active migration of various
family history (62,2 ± 5,8 pg/ml). When comparing patients with types of immune cells to the inflammatory focus, and the period of
EAA and the general group of patients without family history level subacute and chronic allergic inflammation accompanied by marked
of the studied parameters did not reach statistical significance. changes in cytokine production by regulating the cellular immune
Conclusion. Therefore, the results of the study the levels of response in particular IL-12 in patients with EAA.
References:
1. Butler N., Monick M., Yarovinsky T., et al. Altered IL -4 mRNA stability correlates with Th1 and Th2 bias and susceptibility to hypersensitivity pneumonitis in two inbred strains of mice. J Immunol - 2002. - 169: 3700-09.
2. Zacharisen M. C., Schlueter D. P., Kurup V. P., Fink J. N. The longterm outcome in acute, subacute, and chronic forms of pigeon breeder's disease hypersensitivity pneumonitis. Ann Allergy Asthma Immunol - 2002. - 88: 175-82.
3. Fiseselmann J. F., Richerson H. B. Respiratory diseases//Basic and Clinical Immunology. - 8th ed./Ed. by D. P. Stites, Al. Terr, T. G. Parslow. - Norwalk, CT: Appleton & Lange, - 1994. - P. 528-540.
4. Interstitial Lung Diseases/Ed. Olivieri D., du Bois R. M. ERS Monograph. - V. 5. Monograph 14. L., - 2000.
5. Tong Z., Chen B., Dai H., et al. Extrinsic allergic alveolitis: inhibitory effects of pentoxifylline on cytokine production by alveolar macrophages. Ann Allergy Asthma Immunol - 2004. - 92: 234-9.
DOI: http://dx.doi.org/10.20534/ESR-17-1.2-110-113
Ruzibakieva Malika Ruslanovna, Institute of Immunology UzAS, Senior researcher, MD, PhD E-mail: [email protected], Aripova Tamara, Institute of Immunology UzAS, Professor, Director of the Institute of Immunology UzAS,
Hegay Tatyana, Institute of Immunology UzAS Senior researcher, MD, PhD, Kasimov Abdumadjid, Institute of Immunology UzAS, Yuldashev Ulugbek, The Republican Specialized Center of Surgery n. a. Acad. V. Vakhidov
Gender differences in the distribution of HLA II markers in chronic glomerulonephritis and chronic kidney disease in Uzbek population
Abstract: Chronic kidney disease (CKD) leads in the majority of instances to end stage renal disease (ESRD) requiring renal replacement therapy. Our interest was to find the possible associations of HLA class II alleles with ESRD according to the gender specificity in Uzbek population.
Keywords: Chronic kidney disease, End Stage Renal Disease, HLA.
Gender differences in the distribution of HLA II markers in chronic glomerulonephritis and chronic kidney disease in Uzbek population
Chronic kidney disease (CKD) represents an increasing global worldwide health problem particularly in elderly people [1; 2] and/or affected by diabetes, hypertension, and obesity [3]. Therefore, understanding the biological machinery associated with CKD represents an important obj ective in nephrology and internal medicine. According to global WHO data at the moment kidneys diseases are diagnosed for 300-600 million people — about 5-10% of the population of the planet [http://who.int/mediacentre]. According to the Health Institute statistic data in Uzbekistan was observed an increase of patients with End Stage Renal Disease (ERSD) in 2014. The number of reported cases of ESRD in the country was 18000 in 2014, and at the moment there are 1888 patients requiring hemodialysis program.
It is well known that glomerulonephritis progresses to CKD and can develop in ESRD despite receiving specific therapy; the progression level varies and depends on the Host factor. Investigation of the specific genetic-markers in ESRD patients can help to find new strategies of prognosis and prevent ESRD at the early stage. As known, many genome-wide association studies have detected a strong association between the human leukocyte antigen (HLA) and glomerulonephritis over the world populations [4-7], and most of them show the association between HLA and ESRD. The genetic factors play a role not only in the identification of genetic markers of the disease, but also in the emergence of new concepts of the mechanisms of the disease and in addition of information about protective and predisposing factors in the development of diseases, as well as the formation of hypotheses about the totality of exogenous and endogenous factors that lead to disease.
As it is known, for multifactorial diseases has geographic and population conditionally associated with the peculiarities of linkage disequilibrium on different continents, as well for some diseases characterized by a specific distribution by gender, so for example, primary glomerulonephritis occurs twice often in men than in women. Based on the aforesaid, in this study we tried to investigate the association between HLA class II polymorphism and CG, for finding out the mechanisms underlying the initiation and progression of this pathology in Uzbek population according to gender distribution.
The aim of this study is to examine gender differences in the distribution of HLAII gene in patients with chronic glomerulonephritis and chronic kidney disease in the Uzbek population.
Materials and methods
The study was performed at the Institute of Immunology of the Academy of Sciences of Uzbekistan in the lab of human genomics named after Professor R. M. Ruzybakiev and SDC "Immunogen Test" in the period from 2010 to 2016y. y. The selection of patients in outpatient and inpatient treatment was carried out on the basis of RSCS named acad. V. V. Vahidov and SDC "Immunogen Test" at the Institute of Immunology of the AS of Uzbekistan. We have examined 542 people of Uzbek nationality. During the process of selecting individuals for this study we took into account their national identity in three generations, according to the recommendations VII Workshop on HLA (1977). The 225 people of these have been diagnosed with chronic glomerulonephritis, chronic renal failure complicated with end-stage, to whom the chronodialize was recommended. The control group consisted of 317 healthy individuals who are relatives of the first line of kinship. The patients were 51 female and 174 male, mean age was 34,35±11,41. Among the donors were 163 male and 154 female, mean age was 40,84±11,12.
The DNA Isolation was carried out using the method of alcohol-salt treatment by S. Miller et al (1988). HLA-typing of alleles of genes DRB1, DQA1 and DQB1 was performed using kits "of HLA-DNA-TECH" (« DNA Technology SPA», Moscow) using PCR mSSP method (polymerase chain reaction with sequence-specific primers) in modification of Institute of Immunology MOH RF (D.Yu. Trofimov, 1996) and with Q-PCR. As a result of the behavior of the reactions were determined following DRB1 specificity: DRB1 * 01, * 04, * 05, * 07, * 08, * 09, * 10, * 11, * 12, * 13, * 14, * 15, * 16, * 17 (Splits option * 03) * 18 (* Splits version 03). During the process of typing of the DQA1 locus the following specificities were found: * 0101, * 0102, * 0103, * 0201, * 0301, * 0401, * 0501, * 0601. On locus DQB1: * 0201, * 0301, * 0302, * 0303, * 0304, * 0305, * 0401/02 * 0501 * 0502/04 * 0503, * 0601, * 0602-08.
For evaluating of the results obtained, we held the statistical processing of the data, with the help of Arlequin 3.5.2.2 software package, Excel 2007, SISA and a number of formulas to calculate the OR (Odds Ratio) is the index of relative risk, EF (Etiologic fraction) — etiologic fraction, PF (Preventive fraction) — preventive fraction, x2-confidence index Pearson, 95% CI-confidence interval.
Table 1. - The significant alleles of HLA II in men with CRF
HLA Frequencies OR EF PF x2 Pc
CG Male, n=174 Control Male, n =153
DRB1 GF GF
*04 0,0545 0,1732 0,29 2,151 23,079 0,00001
*08 0,0431 0,0098 4,85 0,034 6,815 0,009
*13 0,1925 0,1274 1,75 0,082 5,238 0,02
*14 0,0373 0,0065 4,18 0,028 6,966 0,008
DQA1
*01:03 0,1982 0,1045 2,123 0,104 11,483 0,0007
*03:01 0,1091 0,2254 0,432 1,147 15,368 0,0001
DQ.B1
*02:01 0,1752 0,2352 0,675 0,442 4,209 0,04
*05:03 0,0316 0,0065 0,961 0,04 5,774 0,01
*06:01 0,0775 0,0359 13,20 0,071 5,134 0,023
Note: GF - gene frequency; X - a reliability indicator according to Pearson; OR - Odds ratio; EF - etiologic fraction; PF - preventive fraction, P - a significance indicator by exact Fischer criterion, Pc- a significance indicator across Bonferroni.
Results
As a result of our study have been identified differences in the distribution of alleles' frequencies of genes under study not only in patients and healthy individuals, but also at the separation of the study sample by gender. As can be seen from Table 1, in this sample from the male subjects met all 5 significant markers of predisposition and 4 protecting variants.
The highest relative risks for men have been identified for specific HLA DRB1 * 08 (OR = 4,85), both alleles (OR = 1,75) DRB1 * 14 (6) Splits option DR6 DRB1 * 13 (6) (OR = 4, 18), DQB1 * 06: 01 (OR = 13,2).
In addition to markers of predisposition also met the protecting alleles, the most significance had allelic variants of HLA DRB1 * 04 and DQA1 * 0301.
Then the group offemale patients with CG and CRF was analyzed (Table 2). There were also identified marker alleles, but they differed significantly from the relevant alleles detected in males. It is worth noting that women identified only three predisposing marker 6 and tread options. For most indicators ofthe relative risk for women were identified for both alleles of Splits version DR2 DRB1 * 15 (2) (OR = 2,64) DRB1 * 16 (2) (OR = 9,91), DQA1 * 01: 02 (OR = 2.86).
Also we met and protecting alleles had the greatest significance of allelic variants ofHLA DRB1 * 04, HLA DRB1 * 11 and DQA1 * 03: 01.
Table 2. - Significant alleles of HLA II in women with chronic renal failure
HLA Frequencies OR EF PF x2 Pc
CG Female n=51 Control Female, n=122
DRB1 GF GF
*04 0,0588 0,1352 0,3996 1,380 4,201 0,04
*11 0,049 0,127 0,3541 1,674 4,698 0,03
*15 0,2156 0,0942 2,6423 0,134 9,373 0,002
*16 0,0392 0,004 9,9183 0,035 6,229 0,01
DQA1
*01:02 0,2549 0,1065 2,8684 0,166 12,395 0,0004
*03:01 0,0686 0,1926 0,3088 1,939 8,397 0,003
*05:01 0,1862 0,2786 0,5924 0,609 3,264 0,07
DQB1
*03:01 0,1568 0,2581 0,5345 0,7661 4,192 0,04
*06:02-08 0,2549 0,1434 2,0428 0,130 6,154 0,01
Note: GF - gene frequency; X - a reliability indicator according to Pearson; OR - Odds ratio; EF - etiologic fraction; PF - preventive fraction, P - a significance indicator by exact Fischer criterion, Pc- a significance indicator across Bonferroni.
Thus, as can be seen from the results obtained (Table 3), there parative analysis with the results published earlier, where total samare very interesting gender differences in the distribution of genes ple involving analyzed of both genders, it becomes apparent the HLA II class in patients with CG and CRF, and if you make a com- need to analyze with the obligatory account of gender.
Table 3. - The comparative analysis of significant markers in chronic glomerulonephritis and chronic renal failure in the total group and in the distribution by gender
Total group, (n = 208) [Ruzibakieva M. R. et al. 2016] Male, (n=174) Female, (n=51)
Markers of predisposition DRB1*08, DQA1*01:01, DQA1*01:02, DQA1*01:03, DQB1*04:01, DQB1*05:03, DQB1*06:01, DQB1*06:02-08 DRB1*08, DRB1*13, DRB1*14, DQA1*01:03, DQB1*06:01 DRB1*15, DRB1*16, DQA1*01:02, DQB1*-
Markers of resistance DRB1*04, DQA1*03:01 DQB1*02:01, DQB1*03:02 DRB1*04 DQA1*03:01 DQB1*02:01, DQB1*05:03 DRB1*04, DRB1*11, DQA1*03:01, DQA1*05:01, DQB1*03:01, DQB1*06:02-08
The association of HLA alleles could be used as a risk classification marker and thereby progression to much more severe disease could be protected or diagnosed earlier. This would require further investigation with larger number of patients and healthy in-
dividuals. Also this investigation shows the important role of gender differentiation in prognosis of possible genetically determinate disease appearance or protection against it.
References:
1. Prevalence of chronic kidney disease and decreased kidney function in the adult US population: Third National Health and Nutrition Examination Survey./Coresh J., Astor B. C., Greene T., Eknoyan G., Levey ASAm J/Kidney Dis. - 2003 Jan; - 41 (1):1-12.
2. ESRD patients in - 2004: global overview of patient numbers, treatment modalities and associated trends./Grassmann A., Gioberge S., Moeller S., Brown G/Nephrol Dial Transplant. - 2005. Dec; - 20 (12):2587-93.
3. United States Renal Data System. USRDS Annual Data Report: Epidemiology of Kidney Disease in the United States. Bethesda, Md, USA: National Institute of Diabetes and Digestive and Kidney Diseases; - 2015.
4. Risk HLA-DQA1 and PLA (2)R1 alleles in idiopathic membranous nephropathy./Stanescu H.C., Arcos-Burgos M., Medlar A., Bock-enhauer D., Kottgen A., et al. N. Engl J. Med 364: 616-626; - 2011.
5. A genome-wide association study in Han Chinese identifies multiple susceptibility loci for IgA nephropathy./Yu XQ, Li M., Zhang H., Low HQ. Wei X., et al. Nat Genet - 44: 178-182; - 2011.
Influence of groups of risk on processes of free radical oxidation in blood and urine in the workers of oil and gas refining industry
6. Genome-wide association study identifies susceptibility loci for IgA nephropathy./Gharavi A. G., Kiryluk K., Choi M., Li Y., Hou P., et al. Nat Genet - 43: 321-327; - 2011.
7. HLA has strongest association with IgA nephropathy in genome-wide analysis./Feehally J., Farrall M., Boland A., Gale D. P., Gut I., et al. J Am Soc Nephrol - 21: 1791-1797; - 2010.
DOI: http://dx.doi.org/10.20534/ESR-17-1.2-113-115
Rustamov Muhammadsultan Sadullaevich, Doctor of Philosophy of Medicine, Tashkent Medical Institute of Postgraduate Education, Scientific Research Institute of Sanitary, Hygiene and Occupational diseases Applicant working for degree a doctor E-mail: [email protected] Khamrakulova Mukkadas Askarovna, Doctor of Medical Sciences, Scientific Research Institute of Sanitary, Hygiene and Occupational diseases Chief of the Therapeutic Department Rustamov Khurshid Sadullaevich, Tashkent Pediatric Medical Institute Physician-Ophthalmologist
Influence of groups of risk on processes of free radical oxidation in blood and urine in the workers of oil and gas refining industry
Abstract: The study of the systems of free radical and microsomal oxidation, energetic metabolism appeared to be rational in the prognostic view for determination of the quantitative measure for activity of the oxidative processes in the body and identification of the group of increased risk at the occupational selection for work connected with effect of toxic substances as well as at choice of individual tactics of prevention and therapeutic correction of the oxidative processes. Keywords: free radicals, oxidation, workers, oil and gas refining industry.
Introduction
Early diagnostics of diseases and estimation ofharmful influence of the factors of the environment are the most important problem both in clinic, and at the decision ofhygienic questions [1; 2; 6; 7]. The successful decision of the given problem will allow to develop new effective methods ofearly diagnostics, will help to carry out directed purposeful, the scientifically proved search of optimum ways for improvement of ways for prevention and increase the body resistance [3; 4; 5; 8]. For decision ofthis tasks there is required development of correct prognostic criteria characterized by high sensitivity, accuracy and informativity.
With the account high informativity of parameters of the process of free radical oxidation with use of registration of chemilumi-nescense of blood and urine we investigated biopower and system of microsomal monooxigenases under influence chemical pollutants.
For this purpose in the erythrocytes of 47 workers at the Shur-tan gas and oil-refining complex, having contact with aromatic carbohydrates, there were studied contents of ATPF, ADP, AMP, activity of Na, K-, Ca, Mg-dependent ATPhases, G-6-PDH (glu-cose-6-phosphatdehydrogenase), hexokynase (HK), Phosphofruk-tokinase (PFK), pyruvatkinase (PK), catalase, peroxydase, superox-idismutase (SOD), glutationperoxidase (GP), glutationreductase GR), glutation-S-transferase (GT), glyceroaldehydphosphatdehy-drogenase (GAPDG). Alongside with it there was studied concentration of Na, K, Mg, Ca and Pn, intensity of chemiluminescence of blood and urine, induced by ions Fe 2+' and the state of cytochrom P-450 dependent monooxigenase system.
The contents of adenyl nucleotides in the erythrocytes was measured with use of standard sets "Test combination АТР « and « Test combination FDP/AMP « of firm « Roche Diagnostics « (Switzerland). Concentration of K and Na was measured with method of flame photometry, and the quantity of Ca, Mg and P11 — with use of kits of firm "BRNO" (Cechy). The state of cytochrom-450 dependent monooxygenase system was evaluated by velocity of demetyliration of administered of the drug antipirine into the body, acetylation — by velocity of asoniazide withdrawal and its acety-lated derivatives.
Materials and methods:
All studied participants were divided into 3 groups in view of intensity of influence chemical pollutants: group 1-11 persons not having contact with chemical pollutants; group 2-14 participants, having contact with chemical pollutants 2-3 times a week — 11 men who are not having of contact with chemical загрязнителями; 2-я — 14 men having contact with chemical загрязнителями by periodicity 2-3 times per week (group of risk); group 3-22 persons, constantly contacting with chemical pollutants in the working process during 3-5 years; group of healthy participants included 20 persons working in the administrative unit in the complex, which in the professional activity did not contact to chemical manufacture.
Results and discussion
The carried out analysis of the individual data has shown, that the signs of influence chemical pollutants are found out in the first years of work in the shop on manufacture of pyromellitic
