Authors: Sokolenko A.A., Sydorchuk L.P., Sokolenko M.O.
Introduction. Abdominal obesity (AO) is one of the major independent risk factors of arterial hypertension (AH). AH and AO are multifactorial polygenic diseases. The peroxisome proliferator activated by gamma 2 receptor (PPAR-g2) plays a pivotal role in the lipid metabolism. PPAR receptors modulate the activity of the renin-angiotensin-aldosterone system (RAAS) through transcriptional control of the synthesis of renin, angiotensin, angiotensin-converting enzyme (ACE) and angiotensin II receptor type 1, linking biological effects of RAAS and PPARs. Therefore, the study of genes polymorphism PPAR-γ2 receptor and ACE allows us to explore in detail their role in the pathogenesis of AH and AO.
Our aim was to analyze the frequency of alleles and genotypes of Pro12Ala polymorphism of PPAR--g2 gene (rs1801282) and I/D polymorphism of ACE gene (rs4646994) in patients with essential AH (EAH) combined with overweight or AO.
Material and Methods. 110 screened patients with EAH I–III stages participated in a prospective study: 56.4 % women, 43.6 % men; mean age – 53.3 ± 6.05 years. Among them we revealed EAH I in 22.7 % cases, EAH II – in 45.45 %, EAG III – 31.8 %; overweight persons – 38.2 %, with AO – 53.6 %. The control group included 50 healthy individuals. Alleles of polymorphic locus were studied by polymerase chain reaction based method. Statistical processing was performed usingthe Statistica® 7.0 software. The differences were considered significant at p < 0.05.
Results. The DD-genotype of ACE gene in EAH patients is observed in 39.1 % cases, which is 2.2 times more than in the control group. In patients with EAH dominated "unfavorable" D allele. DD-genotype is associated with a greater frequency of concomitant coronary artery disease, cerebrovascular disease, left ventricle hypertrophy (LVH) in women, higher levels of systolic blood pressure (SBP), and obliterative arterial disease of lower extremities (OADVE).
ProPro-genotype of PPAR-g2 gene was found in 63.6 % cases, which generally does not differ from the control group (60.0 %). Among patients with EAH dominates Pro-allele by 3.9 times (79.5 % vs. 20.4 % Ala-allele carriers), likewise in the control group by 3.5 times (78.0 % vs. 22.0 %), respectively. ProPro-genotype in patients with EAH associated with significantly more frequent cases of diabetes mellitus type 2, AO, LVH in men, OADVE, inherited AO, higher SBP, diastolic BP, and waist circumferences in men. In patients with EAH I dominates I-allele of ACE gene by 3.8 and 3.23 times and ProPro-genotype of PPAR-g2 gene over AlaAla-genotype by 4.67 and 14 times. Exclusively D-allele and Pro-allele carriers were available among patients with EAH III stage. In patients with EAH with normal body weight and AO 1st degree II genotype prevails over DD-option by 2 and 4 times, respectively. In those with overweight, AO 2nd degree the ProPro-genotype dominated the AlaAla-genotype by 12–17 times; among those with overweight available only D-allele carriers; in patients with EAH AO 3rd degree – there are only ProPro-carriers.
Conclusion. DD-genotype or D-allele of ACE gene is a risk factor for EAH II and III stages (OR = 2.33–4.45, p ≤ 0.014–0.013), excess body weight (OR = 4.10–4.45, p < 0.001), AO (OR = 4.45, p < 0.001) with the lowest probability of EAH I stage (OR = 0.22, p = 0.007).
Key words: arterial hypertension, abdominal obesity, АСЕ gene (I/D), PPAR-g2 (Pro12Ala).
Authors: Ancheva I.A.
Introduction. One ofthe mosturgent problemsof modern obstetricsis the occurrenceof iron deficiencyin pregnant women. The experts assess its prevalence in 20–30 % ofcasesin economicallydeveloped countriesand50–80 % – in developing countries.InUkrainein the recent years,the incidenceof iron deficiency anemia during pregnancy increased from15.0 to 22.6 %. The interest ofresearchersto the problemof IDAdue to the factthat inthispathological conditionincidenceof preeclampsia,preterm labor, polyhydramnios, prematurerupture of membranes,weakness of laborforces and otherpregnancy complicationssignificantly increased.According toexperts, theincrease in the amountof blood lossduring laboris in 10 % ofpregnant women withIDA.This populationof pregnant womenhasalsopostpartumseptic complicationsandhypogalactia. Iron deficiency significantly worsensoverall condition of mother and fetus.
Our aim was to evaluate the gene expression of eNOS in placental tissue of the pregnant women with iron deficiency anemia.
Material and Methods. Research performed at the maternity hospital № 2 (Odessa), Genetic Laboratories "Nadiaya" and pathomorphological department of State Institution "IPOG NAMSU" (Head, Professor, MD Zadorozhna TD, Kyiv). The study involved 100 women in labor; we took placenta samples from them. Thus, the following clinical groups were designed: group 1 – the placenta samples of women with physiological pregnancy and childbirth (n = 20), group 2 – the placenta samples of pregnant women with a history of anemia (n = 40), group 3 – the placenta of women with placental dysfunction and a history of anemia (n = 40). In addition to the general clinical research methods, we used current clinical protocol approved by the order № 782 dated 29.12.2005 “On approval of clinical protocols for obstetric and gynecological care”, further research was conducted analysis of immunohistochemical expression of endothelial nitric synthase (eNOS) in placental tissue. Age of pregnant women ranged from 20 to 35 years (28.5 ± 2.8 years).
Discussion. As a result the level of eNOS gene expression increased by 1.4 folds in patients with IDA, while the combination of IDA and placental dysfunction eNOS gene expression level reduced by 10 folds. We discuss the adaptive nature of changes in eNOS gene expression in iron deficiency amongst pregnant women. There were compared genetic and phenotypic characteristics of the enzyme activity of endothelial nitric oxide synthase in decidual tissue. When comparing detected changes of eNOS gene expression with the phenotypic manifestations we found that the expression of endothelial nitric oxide synthase in patients with dysfunction of the placenta in the cytoplasm synticia villi and fetal capillary endothelium and vascular decidua reduced, and the presence of a combination of placental dysfunction and iron deficiency anemia occured with the paradoxical increase expression of the enzyme. Thus, inhibition of eNOS gene expression is a reflection of the exhaustion of the adaptive capacity of the organism. This circumstance suggests that routine administration of nitric oxide precursor (L-arginine medication) in women with gestational endoteliopatiyi manifestations may not give the desired effect due to the presence of a genetically determined deficiency of endothelial nitric oxide synthase and its products. Besides, when isolated iron deficiency anemia occurs than changes in eNOS gene expression is less evident.
Key words: iron deficiency anemia, pregnancy, nitric oxide synthase, genetics.
Authors: Garbuzova V.Yu.
Introduction. BMP-2, like other bone morphogenetic proteins, plays an important role in the development of bone and cartilage, in osteoblast differentiation. As the calcification of the atherosclerotic plaque is an untoward prognostic factor of the acute coronary syndrome, the polymorphism of gene BMP-2 can be associated with the disease progression.
The aim isto establish the association of allelic variants of BMP-2 gene Ser37Ala polymorphism on some characteristics of the acute coronary syndrome (clinical variants, ECG manifestations, development of complications).
Material and Methods. We used venous blood of 118 patients with ACS (22 % women and 78 % men) aged 40 to 73 years (mean age 55.9 ± 0.89 years) hospitalized in the cardiology department of Sumy City Clinical Hospital № 1. The control group consisted of 234 patients. I performed definition of Ser37Ala polymorphism (rs2273073) of BMP-2 gene using PCR with the following restriction fragment length analysis of the allocation of them by electrophoresis in agarose gel. Restriction endonuclease Hpy99I was used for restriction analysis. Statistical analysis was performed by using the software package SPSS-17. Thus, the significance of differences was determined by the χ2-criterion. The value of P < 0.05 was considered as significant.
Results. There isn’t significant difference in the ratio of genotypes (Ser/Ser, Ser/Ala, Ala/Ala) between patients with anginal form of acute coronary syndrome (ACS) and patients with other forms of ACS (аsthmatic, abdominal, arhythmic and painless). It accounted for 51.5; 30.1 and 18.4 % in the group with anginal form of ACS; and 40.0, 46.7 and 13.3 % – in the group with other forms of ACS. Using the χ2-Pearson criterion, it did not reveal association between the Ser37Ala polymorphism of ВМР-2 gene and the clinical variants of ACS (χ2 = 1.652, Р = 0.438). Analysis of the data about correlation Ser37Ala polymorphism of BMP-2 gene with different clinical variants of ACS showed no association of investigated SNP with different clinical manifestations of this disease (unstable angina, non-Q-myocardial infarction, Q-and QS-myocardial infarction). There is no veracity in the distribution of genotypes considering such risk factors as gender, BMI, hypertension, lipid composition and blood hypercoagulation, diabetes, and obesity. The difference of allelic variants frequency was statistically significant (P = 0.033) in groups with different clinical forms of ACS only in patients, who smoke. Smokers with Ser/Ser genotype were significantly more likely to develop anginal form of ACS. Significant difference in the distribution of patients with unstable angina, non-Q-myocardial infarction, Q-and QS-myocardial infarction was found only in patients with obesity (P = 0.018). I did not find significant difference in the distribution of genotypes between patients with complications and those who have had the disease without complications. However, the association with the development of complications of ACS was for male patients, smokers, patients without DAC and without obesity.
Ambiguity and deficiency of data about relationship Ser37Ala and other polymorphisms of BMP-2 gene with the development of ACS requires further research in this area.
Conclusion: Smoking patients with Ser/Ser genotype BMP-2 gene Ser37Ala polymorphism are significantly more likely to develop anginal form of ACS.
There is significant difference in the distribution of genotypes among individuals with unstable angina, non-Q-myocardial infarction, Q-and QS-myocardial infarction in the group without obesity.
There is a link with the development of complications of ACS for male patients, smokers, patients without atherogenic dislipoproteinemia and without obesity.
Key words: allelic polymorphism, bone morphogenetic protein-2, acute coronary syndrome.
Authors: Piddubna A.I., Chemych M.D.
Our aim was to study distribution character of the allelic variants of cytokines genes in HIV-infected Ukrainians.
Data for the study were DNA samples, received from 200 inhabitants of Ukraine: 78 HIV-infected, 22 HIV-negative individuals from the group of high risk of contamination, 100 healthy blood donors. IL-4 (-590C/T), IL-10 (-592C/A) andTNF-α (-308G/A) genes polymorphisms detection was made with the PCR-RLFP method.
By analysis of frequency of IL-4 gene allelic variants, we discovered that homozygotes by the main allel were the dominant variant. Among people with HIV T/T minor gene carriers were 4.5 more often met in comparison with the control group (p < 0.05) that can prove the tendency to association of the mentioned genotype with infection. Distribution of allelic variants of IL-10 gene promoter region in position -592 is characterized by homozygote dominance by the main gene. Among the individuals with HIV A/A minor allel carriers were 3.4 more often than in the control group (p < 0.05). Individuals with A/A genotype were not identified in group of high risk of virus infection. The abovementioned proves the tendency to association of minor allel carrier state with HIV infection. Theoccurrenceofthehomozygouscombinationoftheallelicvariant G/GofthepromoterofTNF-αis provedtoprevailalmosttwofoldovertheoccurrenceofthevariantG/Aamong all groups. High frequency of heterozygote by the main allel was recorded among the individuals with HIV. Thus, G/A genotype frequency in group of HIV-infected people by 2 and 1.5 times exceeded the appropriate indices of group of high risk of infection and comparison group correspondingly (p < 0.05) that points on the tendency to association of the mentioned variant with infection. We concluded that cytokines genes variationsmaycontributetotheacquisitionofHIVinfectionin the Ukrainians andencourages carrying out further populations studies in this sphere of HIV-infection immunogenetics.
Key words: HIV infection, cytokines, allelic polymorphism.
Authors: Rozymenko I.A.
Introduction. Ectonucleotide pyrophosphatase/phosphodiesterase family member 1 is an enzyme, which is encoded by the ENPP1 gene in humans. This gene is a member of the ecto-nucleotide pyrophosphatase/phosphodiesterase (ENPP) family. The encoded protein is a type II transmembrane glycoprotein comprising two identical disulfide-bonded subunits. This protein has broad specificity and cleaves a variety of substrates, including phosphodiester bonds of nucleotides and nucleotide sugars and pyrophosphate bonds of nucleotides and nucleotide sugars. This protein may function to hydrolyze nucleoside 5' triphosphates to their corresponding monophosphates and may also hydrolyze diadenosine polyphosphates. Mutations in this gene have been associated with іdiopathic infantile arterial calcification, ossification of the posterior longitudinal ligament of the spine, insulin resistance. As the calcification of the atherosclerotic plaque is an untoward prognostic factor of the acute coronary syndrome, the polymorphism of gene ENPP1 can be associated with the disease progression.
The aim was to establish the frequency of allelic variants of the ENPPI gene for K121Q polymorphism in patients of different sexes with acute coronary syndrome (ACS).
Material and Methods. We used venous blood of 118 patients with ACS (22 % women and 78 % men) aged 40 to 73 years (mean age 55.9 ± 0.89 years) who were hospitalized in the cardiology department of Sumy City Clinical Hospital № 1. The control group consisted of 110 patients. Definition of K121Q polymorphism (rs1044498) of ENPPI gene was performed using PCR with the following restriction fragment length analysis of the allocation of them by electrophoresis in agarose gel. Restriction endonuclease Eco47I (AvaII) was used for restriction analysis. Statistical analysis was performed using the software package SPSS-17. Thus, the significance of differences was determined by the χ2-criterion. The value of P < 0.05 was considered as significant.
Results. Using the χ2-Pearson criterion, I did not reveal association between the K121Q polymorphism of ENPPI gene and the development of ACS. Distribution of different types of genotype between patients with ACS and healthy patients did not differ statistically significantly. I pointed out that in patients with ACS value homozygotes for the major allele (K/K) and minor allele carriers (K/Q + Q/Q) were 66.9 and 33.1 %, while in the control group –75.5 and 24.5 %, respectively. Factor P, defined by χ2-Pearson criterion, was equal to 0.157 and indicated a lack of significant difference in the distribution of allelic variants of the gene ENPP1 K121Q polymorphism in patients with ACS and the controls.
The value of the given options polymorphism in females was unreliable in patients with ACS and controls (P = 0.280). The distribution of allelic potions K121Q polymorphism in males also did not differ in comparison to patients with ACS and controls (P = 0.320).
Conclusion. There is no link between the polymorphism K121Q of gene ENPP1 and the acute coronary syndrome in males and females.
Key words: ectonucleotide pyrophosphatase/phosphodiesterase, acute coronary syndrome, allelic polymorphism.
Authors: Kmyta V.V.
The purpose of our investigation was study the connection between Bcl1 polymorphism of glucocorticoids receptor gene (GR) with severity of course of disease and obesity patients with bronchial asthma (BA).
188 patients with BA and 95 almost healthy patients were examined. Body mass index, the pulmonary function test (PFT)were investigated. Determination of Bcl1 polymorphism of 2d glucocorticoids receptor exongene (GR) performed with FleuryI. etal. method with modifications. Statistically results processing were done with using of the SPSS-17programs.
The results of analyses of the genotype distribution by Bcl1 polymorphism GR gene dependingon severity of BA course showed that C/C genotype associated with easy course of disease, meanwhile, G/G with severe course. The analyzes of distributing allelic variations GR depending onforced expiratory volume in 1 second (FEV1) showed more expressed obstructive changes in minor allele carries (C/G+G/G) comparing with homozygotes by main allele. We pointed out that patients with BA with C/C genotype index FEV1 showed 68.6 (95% CI 64.9–7.3), and with G/G genotype – 59.9(95% CI 57.02–62.7).
The analysis of genotype distribution by Bcl1 polymorphism GR gene depending on the level FEV1 with consideration BMI showed that genotype C/C mostly occured in patients with normal body mass and FEV1 in normal limits and in range of 60–80% from appropriate values, and genotype G/G –in patients with obesity and severe disorders FEV1.
Obtained results proved the availability of connection between genotypes by Bcl1 polymorphism GR gene, BMI and degree of disorder PFT in patients with BA.
Key words: bronchial asthma, Bcl1 polymorphism, body mass index, the degree of severity.
Authors: Potapov O.O., Kmyta O.P.
Secondary changes in the brain that occur during the early posttraumatic period remain a major cause of death in patients with severe traumatic brain injury. The purpose of our study was to analyze brain changes over time in patients with severe traumatic brain injury studying the association between the results of computed tomography examinations and the -675 4G/5G polymorphism in the PAI-1 gene.
We examined 119 patients with severe traumatic brain injury. Computed tomography changes of brain tissues in patients with severe traumatic brain injury were investigated. Determination of the -675 4G/5G polymorphism in the PAI-1 genewas performed. Statistical processing of the results was done with programsSPSS-17.
We found an association between posttraumatic computed tomography changes of brain tissues in patients with severe traumatic brain injury and the genotypes for the -675 4G/5G polymorphism in the PAI-1 gene: namely, more evident and accelerated involution of lesions in patients with 5G/5G genotype; more rapid recovery in patients with 4G/4G genotype; confirmed predisposition to developing secondary complications, pathological lesions evolution in the brain, and secondary ischemic complications in patients with 4G/5G genotype for the investigated polymorphism.
The obtained results proved the availability of connection between genotypes -675 4G/5G polymorphism in the PAI-1 gene, secondary complications, pathological lesions evolution in the brain, and secondary ischemic complications in patients with severe traumatic brain injury.
Key words: traumatic brain injury, computed tomography examination of the brain, the -675 4G/5G polymorphism in the PAI-1 gene.
Authors: Prystupa L.N., Psarova O.V.
Our aim was to investigate the level of blood lipids and inflammatory markers in patients with coronary artery disease (CAD) depending on carrier ɛ4-allele ɛ2/ɛ3/ɛ4-apolipoprotein E gene polymorphism.
Material and Methods. The study involved 150 patients with CAD. Gene polymorphism of apolipoprotein E studied by Hixson et. al. (1990). DNA was extracted from whole blood using the sets DIAtom DNA Prep 100 («Isogene», Russia). rs7412 and rs429358 polymorphisms exon 4 was determined by polymerase chain reaction followed by restriction fragment length analysis. We performed enzymatic colorimetric method using a set of reagents "Olveks diagnosticum" (Russia) in accordance with the manufacturer's methods to study total cholesterol, HDL and TG. We identified CRP, IL-6 and TNF-α to assess the presence of systemic inflammatory response of the body. Their quantitative assessment was performed using the kits JSC "VECTOR-BEST" (Russia) and by enzyme immunoassay technique according to the manufacturer. Processing of the results was performed using the statistical analysis package SPSS 17.0
Results. Analysis of the genetic polymorphism of apoE showed that among 150 patients with CAD carriers ɛ4-allele were 37 (24.7%) patients, whereas no carriers – 113 (75.3%). We established that carriers ɛ4-allele were mostly men 28 (75.7%), while women – 9 (24.3%); statistically significant differences in Pearson χ2 criterion were found (p = 0.566). We found that in patients who are not carriers of allele frequency ɛ4-hypertension, SBP and DBP levels were significantly higher compared with native ɛ4-allele (p = 0.007, 0.058, 0.03, respectively). The analysis of HDL levels in patients with CAD depending on carrier ɛ4-allele showed that patients who are carriers of ɛ4-allele HDL was significantly lower compared with non-carriers ɛ4-allele (p = 0.013). Atherogenic index (AI) was significantly higher in patients who are carriers ɛ4-allele compared with non-carriers ɛ4-allele (p = 0.006). Patients who are carriers ɛ4-allele had higher levels of CRP and IL-6, but statistically significant differences in the Pearson χ2 criterion were found (p = 0.295, 0.492, respectively).
Conclusion. ɛ4-allele carriers have higher levels of blood lipids and inflammatory markers, indicating the progression of atherosclerosis.
Key words: coronary heart disease, apoE gene polymorphism, lipid profile, inflammatory markers.