[Determination of 2,4-dimethylhydroxybenzene by chromatographic methods in forensic toxicological research of biological material]. / Opredelenie 2,4-dimetilgidroksibenzola khromatograficheskimi metodami pri khimiko-toksikologicheskom issledovanii biologicheskogo materiala.

OBJECTIVE: Is to develop a method for determining 2.4-dimethylhydroxybenzene (2.4-DMHOB) in biological material. The analytical methods used in the experiments were extraction, column chromatography of normal pressure, TLC, GC-MS and HPLC. To extract the analyte from the bioactive matrix, maceration with the binary insulating agent acetone-ethyl acetate (3:7) was used, observing the 2:1 mass ratio of the «insulating agent-matrix¼. Optimal conditions of semi-preparative analyte chromatography were achieved in column (150×10 mm) of «Silasbor¼ S-18 sorbent in elution with a mixture of acetonitrile-water (7:3), which was used in the proposed cleaning scheme, combining extraction and reversed-phase column chromatography. The application of the mobile phase of tetrachloromethane-dioxane (9.5:0.5) has been substantiated for the selective determination of 2.4-DMHOB by TLC («Sorbfil¼ plates). The expediency of confirming identification of the analyte in the form of 2.4-dimethyltrymethylsilylphenol using GC-MS (DB-5MS EVIDEX column (25.000×0.2 mm), stationary phase (5%-phenyl)-methylpolysiloxane, carrier gas - helium) has been shown. The group of characteristic particles in the mass spectrum of trimethylsilyl analyte derivative was represented by 45; 59; 73; 82; 91; 105; 119; 135; 149; 163; 179; 194 m/z ions. HPLC (Discovery C18 250×4.6 mm column, eluting liquid - acetate buffer solution with pH 5.5 - acetonitrile, 50:50) was used to confirm the identification and quantification of 2.4-DMHOB. A method for determining 2.4-DMHOB by the HPLC method in biological material (liver tissue) is proposed, which corresponds to the criteria of linearity, selectivity, accuracy, precision and stability. The minimum detectable quantity of 2.4-DMHOB in the bioactive matrix is 0.5 µg/g, the minimum determined quantity is 1.2 ug/g.

A novel impedimetric sensor based on N-acetyl-L-cysteine for the determination of hydroxyl radicals in cell cultures in vitro.

We report a novel impedimetric sensor based on a graphite electrode impregnated with polyethylene and paraffin under vacuum (IGE) modified with electrochemically deposited gold and a self-assembled monolayer of N-acetyl-L-cysteine (NAC/Au/IGE) for selective and sensitive determination of extracellular hydroxyl radicals (OH•) generated by living cells. The application of a sulphur-containing molecule oxidized by OH• predicts the high selectivity of the sensor, and the utilization of the non-faradaic impedance spectroscopy for recording an analytical response makes it possible to achieve superior sensitivity with a detection limit of 0.01 nM and a linear dynamic range of 0.08-8 nM. Meanwhile, NAC/Au/IGE demonstrated a strong potential of detecting OH• generated by biological objects via successful determination of extracellular hydroxyl radicals generated by normal fibroblast cells and prostate carcinoma cells.

[Study of 2,6-di(propan-2-yl)phenol stability in biomaterial]. / Izuchenie ustoichivosti 2,6-di(propan-2-il)fenola v biologicheskom materiale.

The aim of this study is to research the stability of 2.6-di(propan-2-yl)phenol in biomaterial. GC-MS (column DB-5MS EVIDEX (25 m×0.2 mm); stationary liquid phase of 5%-phenyl-95% dimethylpolysiloxane), TLC (Sorbfil plates, mobile phase of hexane-diethyl ether (9:1) and spectrophotometry (solvent medium - 95% ethanol) were used as methods of analysis. 2.6-di(propan-2-yl)phenol was isolated from the biomatrix (liver tissue) by infusion with a mixture of ethyl acetate-acetone (7:3). The analyte was purified by combining extraction (water-ethyl acetate system) and semi-preparative chromatography on a column of silica gel L 40/100 µm, eluent - hexane-acetone (7:3). It was found that at -22 °C, 0 °C, 12 °C, 20 °C and 30 °C 2.6-di(propan-2-yl)phenol can be present in the liver tissue for 119, 98, 70, 56 and 42 days, respectively. The possibility of mathematical description of analyte decomposition dynamics in biomaterial (liver tissue) at the considered temperatures on the basis of hyperbola equation has been studied. The experimentally calculated coefficients in the hyperbola equation (km) for temperatures -22 °C, 0 °C, 12 °C, 20 °C and 30 °C are equal to 1823, 1130, 697, 510, and 255, respectively. The dependence km on the conserving temperature (tо) was educed. The equation for the description of dependence is offered: km=30.61∙(50-to)-402.39. It is shown that this equation can be the basis for prediction of 2.6-di(propan-2-yl)phenol stability in biomaterial (liver tissue) in the temperature range from -22 °C to 30 °C.

[Propofol: use, toxicology and assay features]. / Propofol: primenenie, toksikologicheskaya kharakteristika i osobennosti opredeleniya.

The study objective is to review the literature on the use, pharmacological properties, toxicology, and assay methods for intravenous anesthetic propofol. The scope and forms of propofol use, its pharmacokinetics, biotransformation features, which occurs more than 90% in the liver, and side effects associated with propofol use for anesthesia, are addressed. Propofol infusion syndrome (also known as PrIS) and deaths from propofol overdose due to medical errors, abuse, suicide attempts, and homicide are reported. Propofol identification and assay methods based on high-performance liquid chromatography (HPLC), gas chromatography with mass spectrometry (GC-MS), and liquid chromatography (LC) are described. The features of the methods performance are outlined; biological materials (the study objects) are listed: mainly blood and plasma, as well as urine, bile, hair, etc. The relevance of a comprehensive forensic chemical study of propofol is indicated, though there are few forensic studies of propofol.

[Study of the stability of 2-methoxy-4-(2-propenyl) hydroxybenzene in biological material]. / Izuchenie ustoichivosti 2-metoksi-4-(2-propenil) gidroksibenzola v biologicheskom materiale.

The purpose of this work is to study the stability of 2-methoxy-4-(2-propenyl)hydroxybenzene [2-MO-4-(2-P)HOB] in biological material. For analysis, gas chromatography-mass spectrometry (GC-MS) was used: column DB-5MS EVIDEX (25 m × 0.2 mm) with a stationary phase of 5%-phenyl-95%-dimethylpolysiloxane; thin layer chromatography (TLC): Sorbfil plates, hexane-dioxane-propanol-2 mobile phase (40:5:1) and UV spectrophotometry (solvent - 95% ethanol). 2-MO-4-(2-P)HOB was isolated from the biomatrix (liver tissue) by infusion with a mixture of ethyl acetate-acetone (7:3). Purification of the analyte was carried out by combining extraction (water-ethyl acetate system) and semi-preparative column chromatography [sorbent - silica gel L 40/100 µm, eluent - hexane-dioxane (8.5:1.5)]. It was established that at -22 °C, 4 °C, 12 °C, 20 °C and 30 °C 2-MO-4-(2-P)HOB is stored in the liver tissue for 385, 357, 301, 245 and 217 days, respectively. We studied the possibility of mathematical description of the dynamics of analyte decomposition in a biomaterial (liver tissue) at the indicated temperatures using the hyperbolic equation. The coefficients in the hyperbola equation (kav), calculated according to the results of the experiment, for temperatures of -22 °C, 4 °C, 12 °C, 20 °C and 30 °C amounted to 6223, 3036, 2387, 1903 and 932, respectively., which is described by the equation: kav=101.19∙(50-to)-1272.78. It was established that on the basis of this equation it is possible to predict the nature of the stability of 2-MO-4-(2-P)HOB in the biomaterial (liver tissue) at temperatures in the range from -22 °C to 30 °C.

[Features of assay and decomposition dynamics of 2-methoxy-4-(1-propenyl)hydroxybenzene in biological material]. / Osobennosti opredeleniya i dinamiki razlozheniya 2-metoksi-4-(1-propenil)gidroksibenzola v biologicheskom materiale.

The objective was to study the features of assay and dynamics of decomposition of 2-methoxy-4-(1-propenyl)hydroxybenzene in biological material. Extraction, semi-preparation chromatography, TLC, HPLC, GC-MS and UV-spectrophotometry were used as test methods. 2-Methoxy-4-(1-propenyl)hydroxybenzene was extracted from the biological material by double infusion (45 min each) with ethyl acetate at a 2:1 mass ratio of isolating agent and biomatrix. Purification was performed by extraction and chromatography in a semi-preparative (190×10 mm) L 40/100 µm silica gel column using a hexane-dioxane (7:3) eluent. The analyte was determined by TLC methods (Sorbfil plates, hexane-acetone 9:1 as a mobile phase), HPLC [Discovery C18 HPLC Column (250×4.6 mm), acetonitrile-acetate buffer pH 5.5 (5:5) as a mobile phase], GC-MS [DB-5MS EVIDEX (25 m×0.2 mm) column with 5%-phenyl-95% dimethyl polysiloxane as a stationary phase], UV-spectrophotometry (95% ethanol as a solvent). The proposed assay method for 2-methoxy-4-(1-propenyl)hydroxybenzene in biomaterial (liver tissue) is validated for linearity, selectivity, accuracy and precision. The study results showed that the decomposition rate of the analyte increases as the store temperature increases. At 0-2 °C, 8-10 °C and 18-22 °C 2-methoxy-4-(1-propenyl)hydroxybenzene is stable for 480, 390 and 260 days respectively.

[Development of methods for determining and studying the persistence of 2,4-dimethylhydroxybenzene and 2,6-dimethylhydroxybenzene in biological material]. / Razrabotka metodik opredeleniya i izuchenie sokhranyaemosti 2,4-dimetilgidroksibenzola i 2,6-dimetilgidroksibenzola v biologicheskom materiale.

OBJECTIVE: To study the features of the determination and preservation of 2.4-dimethylhydroxybenzene and 2.6-dimethylhydroxybenzene in biological material. Extraction, semi-preparative chromatography, TLC, GC-MS and UV spectrophotometry are considered as methods of analysis. The 2.4- and 2.6-dimethylhydroxybenzenes were isolated from the biomaterial by double infusion (30 minutes each) with a mixture of ethyl acetate-acetone (7: 3) at a weight ratio of the insulating liquid and biomaterial of 2:1. Purification was carried out by extraction and chromatography in a semi-preparative (190×10 mm) column of silica gel L 40/100 µm using the eluent hexane-dioxane-propanol-2 (80: 5: 1). Analytes were determined by TLC (Sorbfil plates, mobile phase hexane-dioxane-propanol-2 (120: 5: 1)), GC-MS (DB-5MS EVIDEX column (25 m × 0.2 mm) with a stationary phase (5%-phenyl) - methylpolysiloxane), UV spectrophotometry (solvent - 95% ethanol). The developed methods for the determination of 2.4- and 2.6-dimethyl derivatives of hydroxybenzene in biomaterial (liver tissue) are validated according to the criteria of linearity, selectivity, correctness and precision. The study of the dynamics of decomposition of 2.4- and 2.6-dimethyl hydroxybenzene derivatives in model mixtures with liver tissue, carried out using the developed techniques showed that with an increase in temperature the duration of preservation of analytes in biological material decreases. Moreover, the 2.4-isomer is more stable during storage than the 2.6-isomer. At temperatures of -25 °C, 0-2 °C, 8-10 °C, 20-22 °C, 36 °C the duration of retention of 2.4-dimethylhydroxybenzene is 402, 379, 358 and 224 days, respectively, the duration of retention of 2.6-dimethylhydroxybenzene is 356, 312, 224 and 136 days, respectively.

[Features of determination of 2-methoxyhydroxybenzene in the study of biological material]. / Osobennosti opredeleniya 2-metoksigidroksibenzola pri issledovanii biologicheskogo materiala.

The aim of the study was to develop a method for the determination of 2-methoxyhydroxybenzene in biological material. TLC, UV spectrophotometry, HPLC and GC-MS were used in the experiments. The use of a mixture of ethyl acetate-acetone (7:3 by volume) for the isolation of 2-methoxyhydroxybenzene from biological material is justified. Optimal isolation conditions are established. Purification of the substance was carried out by extraction and chromatography in sorbent column (KCC-3) 80/120 µm. For preliminary identification, TLC was used on Sorbfil PTSX-AF-A-UV plates. Confirmation of identification was carried out by the UV spectrum in ethanol by HPLC with the retention time in a 250×4.6 mm column «SunFire C18¼ (mobile phase acetonitrile-0.025 M potassium dihydrogen phosphate solution 6: 4). Confirmation identification and quantification was performed by GC-MS using a fixed phase capillary column of 5% phenyl-95% methyl polysiloxane after derivatization of the analyte with N-trimethylsilyl-N-methyl trifluoroacetamide (heating for 30 min at a temperature of 60 °C). Ions 45, 58, 73, 91, 107, 136, 151, 166, 181, 196 m/z are present in the mass spectrum of the derivative. The validation of the methodology for the determination of 2-methoxyhydroxybenzene in biomaterial based on the application of the GC-MS method was carried out. The compliance of the methodology with the criteria of linearity, selectivity, correctness, precision and stability is established. The detection limit and the limit of quantification are 8 and 15 µg per 100 g of biomaterial, respectively.

[Association of rs3025058 polymorphism with the development of stroke in patients with cardiovascular pathology].

AIM: To study the association of single-nucleotide polymorphismrs3025058(5а/6а) with the development of stroke in patients of the East Siberian population with cardiovascular pathology and risk factors for its development. MATERIALS AND METHODS: The study involved 260 patients with stroke (age [57.0; 51.062.0]) and 272 patients of the control group (age [55.0; 51.062.0]). Among the patients who underwent stroke, 157 men and 103 women. The control group included 170 men and 102 women. The examination of the main group included: collection of complaints, anamnesis, clinical examination, computed tomography of the brain, electrocardiography, echocardioscopy, ultrasound duplex scanning of the extracranial brachiocephalic arteries, 24-hour monitoring of blood pressure and heart rate, analysis of the blood coagulation system. The patients of the main group had the following cardiovascular pathology and risk factors: arterial hypertension, paroxysmal supraventricular tachycardias, dyslipidemia, atherosclerosis of extracranial brachiocephalic arteries, disorders of the hemostasis system. The control group was examined within the framework of the international project HAPIEE. Molecular genetic research was carried out by real-time PCR. Statistical processing of the material was carried out using the Statistica for Windows 7.0, Excel and SPSS 22 application software. RESULTS: The study established statistically significant associations between the 5a/5a genotype and the 5a allele and stroke in the general group of patients, as well as in the subgroup of men, subgroups of patients with extracranial brachiocephalic arteries atherosclerosis and dyslipidemia. In the subgroup of patients with cardiac arrhythmias, statistically significant results were obtained only for allele 5a, and in the subgroup of women with stroke, subgroups of patients with arterial hypertension and hypercoagulation, no significant associations ofrs3025058(5a/6a) polymorphism with stroke were found. CONCLUSION: Genotype 5a/5a and allele 5a of the single-nucleotide polymorphismrs3025058(5а/6а) increase the risk of stroke in individuals from the East Siberian population, including those in the presence of such risk factors as extracranial brachiocephalic arteries atherosclerosis and dyslipidemia.

Putative role of Brugada syndrome genes in familial atrial fibrillation.

OBJECTIVE: Familial atrial fibrillation (FAF), a not uncommon arrhythmia of the atrium, is characterized by heritability, early onset and absence of other heart defects. The molecular and genetic basis is still not completely clear and genetic diagnosis cannot be achieved in about 90% of patients. In this study, we present the results of genetic screening by next generation sequencing in affected Russian families. PATIENTS AND METHODS: Sixty subjects (18 probands and 42 relatives) with a clinical diagnosis of FAF were enrolled in the study. Since AF frequently associates with other cardiomyopathies, we included all genes that were known to be associated with these disorders at the time of our study. All probands were therefore systematically screened for 47 genes selected from the literature. RESULTS: Our study revealed that seven variants co-segregated with the clinical phenotype in seven families. Interestingly, four out of six genes and three out of seven variants have already been associated with Brugada syndrome in the literature. CONCLUSIONS: To our knowledge, this is the first report of association of the CACNA1C, CTNNA3, PKP2, ANK2 and SCN10A genes with FAF; it is also the first study in Russian families.

[Polymorphisms of the SCN10A Gene in Patients With Sick Sinus Syndrome].

PURPOSE: To study association of rs6795970 polymorphism of SCN10A gene with development of idiopathic sick sinus syndrome (ISSS). MATERIALS AND METHODS: We examined 109 patients with ISSS, 59 their healthy 1­st-, 2­nd-, and 3­rd-degree relatives, and 630 controls. Patients with ISSS were divided into subgroups according to gender and clinical variant of the disease. All patients underwent cardiologic examination and molecular genetic testing of DNA. RESULTS: We revealed significant preponderance of homozygous genotype with rare allele of the studied gene among patients with ISSS compared with control group. In addition, this genotype significantly prevailed among men with SSSU in comparison with the control group. CONCLUSION: Genotype AA of the SCN10A gene is associated with a predisposition to the development of ISSS.

[The Role of the I/D Polymorphism of the ACE Gene in the Development of Atrial Fibrillation].

OBJECTIVE: To study associations of I / D polymorphism of the ACE gene with risk of atrial fibrillation (AF) with the aim of detecting groups of patients prone to development of this disease. MATERIALS AND METHODS: We examined 90 probands with confirmed diagnosis of AF and 144 their I, II, III degrees relatives. These families constituted a core group of our study. The control group comprised 100 relatively healthy people without history of cardiovascular diseases. Methods used in all patients included clinical examination, electrocardiography, echocardiography, Holter ECG monitoring, veloergometry, transesophageal left atrial pacing, molecular-genetic tests. RESULTS: We found statistically significant predominance of genotype II homozygous carriers among probands with primary AF compared with the control group (30.0±7.2 % and 14.0±3.5 %, respectively; p=0.028). Homozygous carriers of DD genotype statistically significantly prevailed in the control group compared with group of probands with primary AF (36.0±4.8 % and 15.0 %±5.6 %; p=0.014). Carriers of homozygous genotype II for common allele statistically significantly prevailed among probands with secondary AF compared with the control group (34.0±6.7 % and 14.0±3.5 %, respectively; p=0.004). Homozygous carriers of DD genotype for the rare allele statistically significantly prevailed among control subjects compared to probands with secondary AF (36.0±4.8 % and 10.0 %±4.2 %, respectively; p=0.001). CONCLUSION: Thus, compared with controls statistically significant preponderance of carriers of homozygous genotype II for common allele was found among probands with both primary and secondary AF. At the same time compared with probands there was a statistically significant predominance of homozygous carriers of DD genotype for the rare allele in the control group. Our findings suggest the heterogeneous nature of AF and confirm that DD genotype homozygosity can be protective against the development of AF.

Polymorphisms of the SCN10A Gene in Patients With Sick Sinus Syndrome.

PURPOSE: To study association of rs6795970 polymorphism of SCN10A gene with development of idiopathic sick sinus syndrome (ISSS). MATERIALS AND METHODS: We examined 109 patients with ISSS, 59 their healthy 1-st-, 2-nd-, and 3-rd-degree relatives, and 630 controls. Patients with ISSS were divided into subgroups according to gender and clinical variant of the disease. All patients underwent cardiologic examination and molecular genetic testing of DNA. RESULTS: We revealed significant preponderance of homozygous genotype with rare allele of the studied gene among patients with ISSS compared with control group. In addition, this genotype significantly prevailed among men with SSSU in comparison with the control group. CONCLUSION: Genotype AA of the SCN10A gene is associated with a predisposition to the development of ISSS.

[Role of AGTR1 A/C polymorphism in the development of atrial fibrillation]. / Rol' polimorfizma A/C gena AGTR1 v razvitii fibrilliatsii predserdii.

AIM: To investigate the AGTR1 A/C polymorphism associated with atrial fibrillation (AF) to form risk groups among patients who are prone to this disease. SUBJECTS AND METHODS: 90 probands with a confirmed diagnosis of AF and their 144 first-, second-, and third-degree relatives were examined. These families made up a study group. A control group was formed of 100 apparently healthy individuals without a history of cardiovascular diseases. Collection of medical history data and complaints, electrocardiography, electrocardiogram monitoring, as well as molecular genetic analysis, thyroid hormone tests were done in all the patients. RESULTS: No statistically significant data on the correlation between the AGTR1 A/C polymorphism and the development of AF were obtained in any patient subgroup. The obtained results can be due to the genetic features of a Siberian population, which are dependent on climatic conditions and geographical location, and confirm that AF is a heterogeneous disease. CONCLUSION: There were no statistically significant differences between the patients in the study group and those in the control group. Our findings suggest the heterogeneity of AF and confirm its multifactorial nature.

TNFR1 -383 A˃C polymorphism and ankylosing spondylitis in a Russian Caucasian population: a preliminary study.

The aim of this study was to assess the association between the TNFR1 rs2234649 polymorphism and ankylosing spondylitis susceptibility in a Russian Caucasian population. A total of 41 ankylosing spondylitis patients and 43 healthy controls, matched according to age and sex, were enrolled, and polymerase chain reaction-restriction fragment length polymorphism analysis was used to genotype the rs2234649 variant. We evaluated genotype distributions in the patient and control groups with the chi-square test, and assessed the relationship between genotypes and ankylosing spondylitis using the odds ratio. Our analysis showed that the rs2234649 polymorphism does not increase ankylosing spondylitis risk. In conclusion, the TNFR1 gene polymorphism tested does not appear to be useful for assessing predisposition to this disease or for its diagnosis or prognosis.

[The Role of Transcriptional Factor Gene TBX5 in Development of Disorders of Cadiac Conduction].

In order to study relationship between development of idiopathic atrioventricular (AV) and intraventricular disorders of cardiac conduction (DCC) with single nucleotide polymorphism (SNP) of TBX5 gene we examined 260 persons with primary DCC (71 patients with abnormal AV conduction, 84 and 105 patients with disordered conduction along right and left brunches of His bundle, respectively) as well as 257 individuals without cardiovascular diseases (control group). Patients were divided into subgroups depending on nosology, age, and sex. Diagnosis was verified by standard cardiological methods and retrospective analysis of available results of previous examinations. Molecular-genetic study of DNA was used for identification of genotype of TBX5 gene SNP. The results indicated significant preponderance of rare GG genotype (CNP-marker rs3825214) of TBX5 gene in the group of patients with left bundle branch block and in the subgroup of women with this pathology. These data suggest that presence of GG genotype (rs3825214) of TBX5 gene increases probability of development of idiopathic DCC along left bundle branch mainly in women.

[Polymorphic alele variants of eNOS gene in patients with disorders of cardiac conduction].

We studied the role of endothelial nitric oxide synthase gene polymorphism 4a/4b in development of such disturbances of cardiac conduction as atrioventricular (AV) block and sick sinus node syndrome (SSNS). We examined 69 subjects (36 men, 33 women) with AV block and 90 subjects (33 men and 57 women) with SSNS. They were divided into groups in dependence on type of conduction disorder and sex. Probands with pathologies studied composed separate groups. All participants underwent included clinical-instrumental cardiological examination and molecular genetic study of eNOS gene polymorphisms. In all groups we revealed significant predominance of a rare homozygous genotype 4b/4b and tendency to decreased number of carriers of widely spread 4a/4a allele.

[The role of alfa-2-beta-adrenoreceptor in development of ventricular conduction disturbance].

BACKGROUND: The purpose of this study was to investigate association between the genetic polymorphism I/D of gene α2ß-adrenoreceptor (ADRA2B) and hereditary disorders of ventricular conduction. PATIENTS AND METHODS: In this study, 102 people with complete left bundle branch block (45.71 ± 1.852 years)--46 females and 56 males, and 86 people with complete right bundle branch block (34.59 ± 1.86 years)--41 females and 45 males. The study was approved by Ethic Committee of the KrasSMU. All participants were included in the study after written informed consent form. Cardiological examination included clinical examination, electrocardiography, echocardiography, Holter monitoring, stress-test, koronaroangiografy and radionuclide method of a myocardium and molecular and genetic researches. RESULTS: Statistically, significant prevalence of a homozygous genotype of DD on rare allele gene ADRA2B in both groups in comparison with group of control is established. The reliable dominance of the homozygous rare genotypes (D allele) of gene ADRA2B were detected in all groups. CONCLUSION: Polymorphism DD of a gene ADRA2B is a genetic predictor of predisposition to the blockade of the right and left bundle branch block.

[α-2ß-adrenoreceptor gene polymorphism in patients with disorders of cardiac conduction].

The article is devoted to the role of insertion-deletion polymorphism of -2-adrenoreceptor gene in development of hereditary disorders of cardiac conduction. We examined 71 patients with atrioventricular blocks and 92 patients with sick sinus node syndrome. Statistically significant preponderance of homozygous genotype DD of ADRA2B gene was found in both groups. Associations of alleles with male or female gender were also revealed.