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Research ArticleBeta2-Adrenergic Receptor Gene Polymorphisms in EgyptianPatients with Acute Myocardial Infarction

Eman Toraih1 Mohammad H Hussein2 and Dahlia I Badran3

1Genetics Unit Department of Histology and Cell Biology Faculty of Medicine Suez Canal University Ismailia 41522 Egypt2Department of Chest Diseases Faculty of Medicine Cairo University Giza Egypt3Department of Medical Biochemistry Faculty of Medicine Suez Canal University Ismailia 41522 Egypt

Correspondence should be addressed to Eman Toraih emantoraihgmailcom

Received 31 July 2014 Revised 8 November 2014 Accepted 17 November 2014 Published 9 December 2014

Academic Editor Piero R Bianco

Copyright copy 2014 Eman Toraih et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Background Beta2-adrenergic receptor (ADRB2) gene polymorphisms Arg16Gly and Gln27Glu have been implicated in the

pathogenesis of cardiovascular diseases The aim of this study was to determine the association of these two polymorphisms withthe risk of myocardial infarction (MI) in the Egyptian populationMethods Blood samples were collected from 68MI patients and75 healthy controls They were assessed for the presence of cardiovascular risk factors and genotyped for the Arg16Gly (rs1042713)and Gln27Glu (rs1042714) polymorphisms using allelic-discrimination polymerase chain reaction Results There is no significantdifference in genotype and allele frequencies at codon 16 between MI patients and controls (119875 = 0919) However at codon 27MI risk was higher in Gln

27homozygous participants than in Glu

27carriers (119875 = 0045) The haplotype frequency distribution

showed significant difference among cases and controls (119875 = 0002) homozygotes for Gly16Gln27haplotype were more susceptible

to MI than Gly16Glu27

carriers Patients with Arg16Gln27

haplotype had higher serum total cholesterol levels (119875 lt 005) andlower frequency of diabetes in MI patients (119875 lt 001) However both Glu

27genotypes and haplotype showed lower frequency of

hypertension (119875 lt 0001) Conclusions Our findings suggested that the ADRB2 gene polymorphisms may play an important rolein susceptibility of MI among Egyptian population

1 Introduction

Myocardial infarction (MI) is a major cause of morbidity andmortality worldwide accounting for up to 40 of all deaths[1 2] It is caused by myocardial cell death due to prolongedischemia [3]MI is amultifactorial polygenic disorder drivenby interactions of an individualrsquos genetic background andseveral environmental factors [4ndash7] Greater understandingof MI etiology is mandatory to identify individuals at highrisk and to improve prevention and therapy of this commonand important condition Cardiovascular risk factors assmoking obesity hypertension dyslipidemia and diabeteshave long been known [8] However these risk factors donot explain why some individuals are more susceptible tothese environmental determinants in comparison to otherswith the same given risk factors Several genomic associationstudies family-based and twin studies and linkage analysisprovide insights into multiple susceptible genes underlyingMI disease [9 10]

The beta 2-adrenergic receptors (1205732-AR) a member of

the G-protein-coupled receptor superfamily have attractedgreat attention due to their multiple physiological and healtheffects particularly those involving cardiovascular pheno-types [11ndash16] In vascular smooth muscle 120573

2-AR medi-

ate vasodilation in response to adrenergic agonists andin healthy myocardium they mediate chronotropic andinotropic responses to endogenous and exogenous adrenergicagents [17ndash21] These functions render 120573

2-AR an important

target for cardiovascular disease (CVD) therapy [15 2223] Interestingly studies reported 120573

2-AR dysfunction to be

implicated in the pathogenesis of several CVD includinghypertension [24ndash27] coronary heart disease [13 23] idio-pathic dilated cardiomyopathy [28] congestive heart failure[29 30] idiopathic ventricular arrhythmias [31] and suddencardiac death [32]

Several single nucleotide polymorphisms (SNP) in the1205732-

AR (ADRB2) gene (MIM 109690) have been identified in

Hindawi Publishing CorporationAdvances in Molecular BiologyVolume 2014 Article ID 471635 11 pageshttpdxdoiorg1011552014471635

2 Advances in Molecular Biology

many populations [33 34] The most important two SNPsare located within the amino-terminal extracellular domainnear the receptorrsquos ligand-binding site [35] The first SNPwas caused by a change of adenine to guanine (A gt G)at nucleotide 46 resulting in amino acid substitution fromarginine to glycine at codon 16 (Arg16Gly or R16G) Thesecond SNP is the change of cytosine to guanine (C gt G)at nucleotide 79 causing the substitution from glutamineto glutamic acid at codon 27 (Gln27Glu or Q27E) [15 3637] In vivo and in vitro functional studies suggested thatthese two polymorphisms have altered receptor functionand behavior [38ndash42] Several clinical and pharmacologicalstudies highlighted the important role of these polymor-phisms in the pathogenesis of CVD and their impact on drugresponse [22 30 39 43] However evidence regarding therole of ADRB2 gene polymorphisms in patients with MI isambiguous and inconsistent Significant differences in thegenotype frequencies and haplotype structure ofADRB2 genewere noted in different ethnic populations [13 23 25 44 45]Thus this case-control study was conducted to determine theprevalence of these ADRB2 gene polymorphisms in Egyptianpopulation and to assess their genetic association with thesusceptibility to MI disease

2 Materials and Methods

21 Study Participants The study population was composedof 68 patients with acute myocardial infarction (MI) and 75age-matched healthy controls with no history of cardiovas-cular events hypertension or any other chronic diseasesPatients were recruited from Coronary Care Unit (CCU)Suez Canal University Hospital Ismailia Egypt during theperiod between April 2013 and March 2014 The diagnosisof MI was confirmed by evidence of symptoms in thepresence of either diagnostic elevations of cardiac enzymesor diagnostic changes on electrocardiograms (ECG) [46]Thecardiovascular risk factors were assessed in all participantsTheyunderwent evaluations includingmeasurement of bloodpressure body mass index (BMI) fasting serum lipid andglucose levels cardiac enzymes and ECG Diabetic patientseither had a known history of diabetes mellitus or were diag-nosed according to the American Diabetic Association rec-ommendations [47] Hypertensionwas defined as the averageSBP ge 140mmHg andor the average DBP ge 90mmHgandor self-reported current treatment for hypertension withantihypertensive medication [48] The study was approvedby the Medical Research Ethics Committee of Faculty ofMedicine Suez Canal University Written informed consentwas obtained from all participants

22 Samples Collection and Biochemical Measurements Fast-ing blood samples were collected from both cases andcontrols Blood from patients was collected within 24 hoursof CCU admission Two milliliters (mL) of venous blood waswithdrawn in EDTA tubes to be used for subsequent geneticanalysis Another two milliliters was collected in VacutainerSerum Separator Tubes II (Becton Dickinson Plymouth)to obtain serum Parameters such as fasting blood sugar

(FBS) total cholesterol (TC) triglycerides (TG) high den-sity lipoproteins-C (HDL-C) and low density lipoprotein-C(LDL-C) were analyzed within 2 h after collection Fastingblood sugar was estimated using RocheHitachi analyzer kitsbased on the glucose oxidase and peroxidase method [49]Serum TC and TG were measured by enzymatic colorimetricdetermination method using RocheHitachi cholesterol kit(Cat number 1489232) [50 51] and RocheHitachi triglyc-erides kit (Cat number 1488872) [52] HDL-C was measuredusing RocheHitachiHDL-C kit (Cat number 03030024) andwas estimated by phosphotungstic acid precipitation followedby enzymatic analysis in supernatant fraction [53] LDL-Cwas calculated by the Friedewald formula after consideringits limitations if TG gt 400mgdL LDL-C = TC minus (HDL-C +TG5) [54] FBS lt 126mgdL TC levels lt200mgdL TG lev-els lt150mgdL LDL-C lt 130mgdL andHDL-C gt 60mgdLwere considered normal [55]

23 Genotyping Genomic DNA was extracted from bloodleukocytes using QIAamp DNA Blood Mini kit (QIAgenClinilab Co Egypt Catalog number 51104) according to themanufacturerrsquos instructions [56] Concentration andpurity ofthe extracted DNA were measured by NanoDrop ND-1000(NanoDrop Tech Inc Wilmington DE USA) Genotypingof the ADRB2 polymorphisms (Arg16Gly rs1042713 andGln27Glu rs1042714) was performed by real-time poly-merase chain reaction (RT-PCR) technology PCR was per-formed in a 25-120583L reaction volume containing 125120583L Taq-man Universal PCR Master Mix No AmpErase UNG (2x)20 ng genomic DNA diluted to 1125 120583L with DNase-RNase-free water and 125 120583L 20 times TaqMan SNP Genotyping AssayMix (Applied Biosystems Egypt assay ID C 2084764 20 andC 2084765 20) Internal positive controls and no-templatecontrols (no DNA) were included in each run PCR ampli-fication was carried out in an AB 7500HT instrumentwith the Sequence Detection System (SDS) Software version211 (Applied Biosystems Egypt) according to the followingconditions initial denaturation at 95∘C for 10min followedby 40 cycles of 92∘C for 15 sec and 60∘C for 1min Genotypingwas performed blinded to casecontrol status Ten per cent ofthe randomly selected samples were regenotyped in separateruns to exclude the possibility of false genotype calls with100 concordance rate

24 Statistical Analysis Statistical analysis was carried outusing the Microsoft Excel 2010 and the ldquoStatistical Packagefor the Social Sciences (SPSS) for windowsrdquo software ver-sion 20 Odds ratios (OR) with a 95 confidence inter-val (CI) were calculated Categorical variables were com-pared using the chi-square (1205942) while Mann-Whitney 119880and Kruskal-Wallis tests were used to compare continu-ous variables Due to significant differences in some car-diovascular risk factor variables between cases and con-trol participants binary logistic regression analysis wasperformed that included the ADRB2 polymorphism andpotential confounders (age gender smoking status familyhistory of cardiovascular disease prevalence of obesity andhyperlipidemia) A two-tailed 119875 value of 005 was con-sidered statistically significant The allele frequency within

Advances in Molecular Biology 3

each group was determined as the number of occurrencesof an individual allele divided by the total number ofalleles The carriage rate was calculated as the number ofindividuals carrying at least one of the investigated allelesdivided by the total number of individuals in each group[57 58] The Hardy-Weinberg equilibrium was calculatedusing the Online Encyclopedia for Genetic Epidemiology(OEGE) software (httpwwwoegeorgsoftwarehwe-mr-calcshtml) and tested by 1205942 test to compare the expectedgenotype frequencies among patient and control groups

3 Results

31 Molecular Analysis of ADRB2 Gene Polymorphisms Atotal of 68 patients and 75 controls were included in theanalysis Characteristics of the study population are shownin Table 1 SNP analysis of Arg16Gly (R16G rs1042713) andGln27Glu (Q27E rs1042714) polymorphisms of ADRB2 geneshowed that the frequencies of Arg

16and Gly

16alleles in

our study population were 037 and 063 while those ofGln27

and Glu27

alleles were 067 and 033 respectively Thedistribution of Arg16Gly genotypes and alleles did not differsignificantly betweenMI patients and controls For Gln27GlupolymorphismGln

27Glu27was significantly themost preva-

lent genotype in the healthy control group (507) whileGln27Gln27

was the most predominant genotype amongpatients (50) (119875 = 0045) The frequency of Glu

27allele

was significantly lower in MI patients compared to controls(331 versus 453 119875 = 0034) see Table 2 The distributionof ADRB2 genotypes among patients and controls was foundin accordance with those expected by the Hardy-Weinbergequilibrium (119875 gt 005) Haplotype analysis revealed thepresence of three haplotypes only Arg

16Gln27 Gly16Gln27

and Gly16Glu27 Their frequencies were 030 029 and 041

respectively Different combinations of haplotypes existedin the study population and showed significant differencebetween MI patients and healthy controls (119875 = 0002) seeTable 3 The frequency of Gly

16Glu27

haplotype was signif-icantly lower in MI patients than controls (119875 = 0045) Inaddition homozygote individuals for Gly

16Gln27haplotype

had higher risk of developing MI compared to noncarrierswith adjusted OR (95 CI) of 204 (14ndash296) see Table 4

32 ADRB2 Gene Polymorphisms and Disease CharacteristicsCharacteristics of patients according to ADRB2 genotypesand haplotypes are demonstrated in Tables 5 and 6 Dia-betes mellitus (DM) frequency was significantly higher inGly16Gly16

genotype carriers (625) compared to othergenotypes carriers (62 and 313 for Arg

16Arg16

andArg16Gly16 resp) (119875 = 0003) The frequency of hyperten-

sion was significantly higher in MI patients with Gln27Gln27

genotype (667) compared to their counterparts (25 forGln27Glu27

and 83 for Glu27Glu27) (119875 lt 0001) In

addition MI patients with the Arg16Gln27

haplotype hadsignificantly higher serum cholesterol levels compared tononcarriers (206 plusmn 36 versus 188 plusmn 42 119875 = 003)

Table 1 Baseline characteristics of the study groups

Variables Patients (119899 = 68) Controls (119899 = 75) 119875 valuelowast

Risk factors for CVDAge y 535 plusmn 74 528 plusmn 61 0203Male gender 45 (662) 53 (707) 0252Smokers 37 (544) 15 (200) lt0001FH of CVD 35 (515) 17 (227) lt0001BMI kgm2 295 plusmn 36 274 plusmn 27 lt0001Hypertension 48 (706) mdashSBP mmHg 1297 plusmn 227 1220 plusmn 112 0010DBP mmHg 838 plusmn 133 775 plusmn 77 0001Diabetic 32 (471) mdashFBS mgdL 1292 plusmn 457 982 plusmn 113 lt0001

Lipid profileHyperlipidemia 62 (912) 38 (506) lt0001TC mgdL 199 plusmn 40 174 plusmn 40 0002TG mgdL 203 plusmn 63 151 plusmn 84 lt0001HDL-C mgdL 38 plusmn 74 474 plusmn 12 lt0001LDL-C mgdL 121 plusmn 41 96 plusmn 40 0001VLDL-C mgdL 405 plusmn 13 302 plusmn 17 lt0001

Cardiac enzymesCK ngmL 1688 plusmn 1832 mdashCK-MB ngmL 320 plusmn 370 mdashLDH IUL 1562 plusmn 1223 mdash

Values are presented as number (percentage) or mean plusmn standard devia-tion Chi-square (1205942) and Mann-Whitney 119880 tests were used lowastStatisticallysignificant at 119875 lt 005 FH family history CVS cardiovascular diseaseBMI body mass index SBP systolic blood pressure DBP diastolic bloodpressure FBS fasting blood sugar TC total cholesterol TG triglycerideHDL-C high density lipoprotein cholesterol LDL-C low density lipoproteincholesterol VLDL-C very low density lipoprotein cholesterol CK totalcreatine kinase CK-MB creatine kinase isoenzyme MB fraction LDHlactate dehydrogenase

4 Discussion

To the best of our knowledge this is the first work examiningthe association of Arg16Gly and Gln27Glu polymorphisms ofbeta2-adrenergic receptor with acute myocardial infarction

in Egyptian population Frequency distribution of allelesand genotypes was in Hardy-Weinberg equilibrium for bothSNPs Gly

16and Gln

27were the most prominent alleles

representing 063 and 067 of the population respectivelyCorrespondingly Gly

16Gly16

and Arg16Gly16

at codon 16and Gln

27Gln27

and Gln27Glu27

at codon 27 were themost common genotypes in the Egyptian population Thiswas in agreement with another Egyptian study where Gly

16

and Gln27

were the most frequent alleles representing 060and 075 respectively [59] Similar frequencies in priorstudies were found in different ethnic populations (AfricanAmerican Caucasian and Saudian populations) [33 35 3960 61] In contrast individuals with Asian descent reportedhigher frequencies of Arg

16and Gln

27alleles and absence of

Glu27Glu27genotype [39 59 60 62ndash64] Haplotype analysis

revealed the presence of 3 haplotypes only their frequencieswere 30 for Arg

16Gln27 29 for Gly

16Gln27 and 41


Table 2 Genotype and allele frequencies of ADRB2 gene polymorphisms in patients and controls

Variables Patients (119899 = 68) Controls (119899 = 75) 1205942

119875 value CrudeOR (95 CI)

AdjustedampOR (95 CI)

Arg16Gly polymorphismGenotype frequency

Arg16Arg16 10 (147) 08 (107) 0010 0919 10 10Arg16Gly16 30 (441) 40 (533) 06 (02ndash17) 03 (007ndash20)Gly16Gly16 28 (412) 27 (360) 08 (02ndash24) 19 (03ndash99)

Allele frequencyArg16 allele 50 (368) 56 (373) 0009 0920 10 02 (01ndash07)Gly16 allele 86 (632) 94 (627) 10 (06ndash16) 08 (02ndash37)

Gln27Glu polymorphismGenotype frequency

Gln27Gln27 34 (500) 22 (293) 4012 0045lowast 10 10Gln27Glu27 23 (338) 38 (507) 03 (01ndash08)lowast 05 (01ndash15)Glu27Glu27 11 (162) 15 (200) 04 (01ndash12) 19 (04ndash82)

Allele frequencyGln27 allele 91 (669) 82 (547) 4459 0034lowast 10 02 (006ndash11)Glu27 allele 45 (331) 68 (453) 05 (03ndash09)lowast 05 (01ndash15)

Values are shown as number () Chi-square (1205942) for trend was used OR (95 CI) odds ratio and confidence interval ampAdjusted for confounding factorsAdjusted OR for alleles was calculated as presence versus absence of this particular allele lowastStatistically significant at 119875 lt 005

Table 3 Haplotype combinations of ADRB2 gene polymorphisms in patients and controls

Haplotype combinations Patients (119899 = 68) Controls (119899 = 75) 1205942119875 value Crude

OR (95 CI)AdjustedampOR (95 CI)

Gly16Glu27 + Gly16Glu27 11 (162) 15 (200) 9404 0002lowast 10 10Gly16Glu27 + Gly16Gln27 5 (740) 13 (173) 05 (01ndash19) 02 (002ndash15)Gly16Glu27 + Arg16Gln27 18 (265) 25 (333) 09 (03ndash26) 02 (005ndash13)Gly16Gln27 + Arg16Gln27 12 (176) 15 (200) 11 (03ndash32) 01 (001ndash08)Arg16Gln27 + Arg16Gln27 10 (147) 6 (800) 22 (06ndash81) 05 (007ndash39)Gly16Gln27 + Gly16Gln27 12 (176) 1 (130) 163 (18ndash145)lowast 90 (05ndash161)Values are shown as number () Chi-square (1205942) for trend was used OR (95 CI) odds ratio and confidence interval ampAdjusted for confounding factorslowastStatistically significant at 119875 lt 005

Table 4 Haplotype frequencies of ADRB2 gene polymorphisms in patients and controls according to the copy number




Arg16Gln27

Negative 28 (412) 29 (387) 0146 0703 10 101 copy 30 (441) 40 (533) 07 (03ndash15) 04 (01ndash166)2 copies 10 (147) 6 (80) 17 (05ndash53) 05 (007ndash39)

Gly16Gln27

Negative 39 (574) 46 (614) 3378 0066 10 101 copy 17 (250) 28 (373) 07 (03ndash14) 03 (01ndash108)2 copies 12 (176) 1 (13) 141 (17ndash113)lowast 204 (14ndash296)lowast

Gly16Glu27Negative 34 (500) 22 (293) 4012 0045lowast 10 101 copy 23 (338) 38 (507) 03 (01ndash08)lowast 05 (017ndash15)2 copies 11 (162) 15 (200) 04 (01ndash12) 19 (047ndash82)

Values are shown as number () Chi-square (1205942) for trend was used OR (95 CI) odds ratio and confidence interval ampAdjusted for confounding factorslowastStatistically significant at 119875 lt 005


Table 5 Disease characteristics of patients according to ADRB2 genotypes (119899 = 68)

Characteristics Arg16Gly genotypes119875 value Gln27Glu genotypes

119875 valueArg16Arg16 Arg16Gly16 Gly16Gly16 Gln27Gln27 Gln27Glu27 Glu27Glu27

Total number 10 (147) 30 (441) 28 (412) 34 (500) 23 (338) 11 (162)Risk factors for CVD

Age at diagnosis 52 plusmn 74 53 plusmn 81 55 plusmn 66 0577 53 plusmn 77 55 plusmn 77 53 plusmn 53 0367Male gender 7 (155) 22 (489) 16 (356) 0412 22 (440) 21 (420) 7 (140) 0059Smokers 4 (108) 20 (541) 13 (351) 0185 18 (487) 15 (405) 4 (108) 0278FH of CVD 7 (200) 16 (457) 12 (343) 0325 21 (600) 10 (286) 4 (114) 0219Hypertension 10 (208) 20 (417) 18 (375) 0085 32 (667) 12 (250) 4 (830) lt0001lowast

DM 2 (625) 10 (313) 20 (625) 0003lowast 14 (437) 11 (344) 7 (219) 0429BMI Kgm2 32 plusmn 38 30 plusmn 40 29 plusmn 36 0080 30 plusmn 33 29 plusmn 41 28 plusmn 29 0195

Lipid profileHyperlipidemia 10 (161) 28 (452) 24 (387) 0336 31 (500) 20 (323) 11 (177) 0455Total cholesterol mgdL 202 plusmn 33 208 plusmn 38 188 plusmn 43 0085 199 plusmn 41 207 plusmn 40 181 plusmn 32 0139Total triglyceride mgdL 228 plusmn 45 200 plusmn 74 195 plusmn 54 0121 199 plusmn 56 199 plusmn 81 218 plusmn 38 0199HDL-C mgdL 38 plusmn 52 39 plusmn 67 37 plusmn 87 0374 38 plusmn 77 39 plusmn 80 36 plusmn 48 0883LDL-C mgdL 119 plusmn 30 1129 plusmn 37 111 plusmn 46 0125 122 plusmn 43 128 plusmn 39 101 plusmn 35 0099VLDL-C mgdL 46 plusmn 89 40 plusmn 15 39 plusmn 11 0121 40 plusmn 11 40 plusmn 16 44 plusmn 75 0199

Cardiac enzymesCK ngmL 1624 plusmn 1422 1499 plusmn 1654 1912 plusmn 2149 0746 1821 plusmn 2088 1717 plusmn 1735 1215 plusmn 1078 0791CK-MB ngmL 362 plusmn 412 328 plusmn 336 294 plusmn 402 0306 294 plusmn 320 450 plusmn 470 126 plusmn 107 0089LDH IUL 1666 plusmn 603 1368 plusmn 1161 1733 plusmn 1437 0289 1775 plusmn 1517 1301 plusmn 882 1451 plusmn 625 0506

Values are shown as mean plusmn standard deviation or number (percentage) CVD cardiovascular disease DM diabetes mellitus BMI body mass index HDL-Chigh density lipoprotein cholesterol LDL-C low density lipoprotein cholesterol CK total creatine kinase CK-MB creatine kinase isoenzyme MB fractionLDH lactate dehydrogenase Two-sided chi-square and Kruskal-Wallis tests were usedlowastStatistically significant at 119875 lt 005

for Gly16Glu27 The absence of Arg

16Glu27

was consistentacross several haplotype studies conducted in different ethnicpopulations including African Americans Brazilians Euro-peans Asians and Hispanic-Latinos populations [35 36 63ndash69] Strong linkage disequilibrium (LD) exists between thesetwo SNPs and as a result the Arg

16allele is typically seen

with Gln27 whereas Gly

16can coexist with either Gln

27or

Glu27[33 59 60 63 70]

In the present study the distribution of Arg16Gly geno-types and alleles did not differ significantly between caseswith MI and control subjects This was in agreement withothers who reported no effect of Arg16Gly polymorphismwith the occurrence of MI disease among Americans Euro-peans and Turkish populations [15 19 25 71] Howeverfor Gln27Glu polymorphism our results showed higherfrequency of the Gln

27variants among patients and that

the Glu27

allele conferred protection against the occurrenceof MI disease This was similar to previous studies whichreported a significant association of Gln

27homozygotes with

coronary events ventricular tachyarrhythmias and suddencardiac death [14 15 27 32 72] Others highlighted theprotective role of Glu

27allele against MI in young and elder

populations [14 19 44 71] Also Kaye et al showed thatsubjects with the Glu

27allele had a significant improvement

in left ventricular ejection fraction compared with subjectshomozygous for Gln

27[73] In contrast one study found

the opposite correlation carriers of Glu27allele had a higher

incidence of coronary artery disease and a higher likelihoodof later need for coronary revascularization [74] while othersdid not reveal such associations with MI [11 13 23 75]

The Arg16Gly and Gln27Glu polymorphisms are knownto alter the functional properties of the receptor and itsbehavior after agonist exposure [38 76] Although thesetwo amino terminal polymorphisms are located near theligand-binding site they do not alter the binding capacity ofendogenous or exogenous catecholamines to 120573

2-AR or affect

further G-protein coupling and adenylyl cyclase activation[35 77] In addition receptor synthesis rates and agonist-promoted internalization were not different between thereceptors [78 79] However studies of agonist stimulation incultured cells demonstrated that the Glu

27receptor exhibited

enhanced resistance to downregulation when compared withthe Gln

27variant as assessed by receptor number [77 80]

Other in vitro studies proposed that this desensitization phe-nomenon might be due to differential alterations in receptordegradation after the internalization step [78 81 82] More-over another difference was demonstrated by cell-signalingstudies in two cell lines the Glu

27variant was associated

with magnification of the catecholamine-induced activationof intracellular signaling transduction [83] thus likely pro-moting superior cardiovascular performance in human [84]

To examine possible additive or synergistic effects ofthe two tested polymorphisms haplotype analysis withinthe ADRB2 locus was done revealing significant difference


Table 6 Disease characteristics of patients according to ADRB2 haplotypes (119899 = 68)

Characteristics Arg16Gln27119875 value Gly16Gln27

119875 value Gly16Glu27119875 value

Positive Negative Positive Negative Positive NegativeTotal number 40 (59) 28 (41) 29 (427) 39 (573) 34 (50) 34 (50)Risk factors for CVD

Age at diagnosis 53 plusmn 79 55 plusmn 66 0355 538 plusmn 78 533 plusmn 71 0582 543 plusmn 71 527 plusmn 77 0443Male gender 29 (644) 16 (356) 0188 16 (356) 29 (644) 0098 25 (556) 20 (444) 0200Smokers 24 (649) 13 (351) 0269 12 (387) 19 (613) 0548 15 (484) 16 (516) 0808FH of CVD 17 (515) 16 (485) 0234 13 (394) 20 (606) 0598 20 (606) 13 (394) 0089Hypertension 30 (625) 18 (375) 0340 24 (500) 24 (500) 0058 16 (333) 32 (667) lt0001lowast

DM 12 (375) 20 (625) 0001lowast 17 (531) 15 (469) 0100 18 (563) 14 (437) 0331BMI Kgm2 302 plusmn 4 285 plusmn 3 0121 291 plusmn 3 297 plusmn 4 0955 289 plusmn 4 300 plusmn 3 0090

Lipid profileHyperlipidemia 38 (613) 24 (387) 0184 25 (403) 37 (597) 0213 31 (500) 31 (500) 1000Total cholesterol mgdL 206 plusmn 36 188 plusmn 42 0030lowast 192 plusmn 44 204 plusmn 36 0070 198 plusmn 39 199 plusmn 41 0731Total triglyceride mgdL 207 plusmn 68 196 plusmn 54 0945 192 plusmn 59 210 plusmn 65 0330 205 plusmn 71 199 plusmn 56 0864HDL-C mgdL 385 plusmn 63 371 plusmn 8 0170 38 plusmn 87 382 plusmn 6 0471 380 plusmn 71 379 plusmn 7 0917LDL-C mgdL 126 plusmn 36 112 plusmn 46 0061 115 plusmn 48 124 plusmn 34 0133 119 plusmn 39 121 plusmn 43 0722VLDL-C mgdL 414 plusmn 13 392 plusmn 11 0945 384 plusmn 11 421 plusmn 13 0330 411 plusmn 14 399 plusmn 11 0864

Cardiac enzymesCK ngmL 1530 plusmn 1583 1912 plusmn 2149 0842 1978 plusmn 2359 1471 plusmn 1305 0995 1554 plusmn 1554 1821 plusmn 2088 0581CK-MB ngmL 337 plusmn 351 294 plusmn 402 0159 327 plusmn 405 314 plusmn 347 0660 345 plusmn 418 293 plusmn 320 0835LDH IUL 1442 plusmn 1051 1733 plusmn 1437 0852 1655 plusmn 1662 1493 plusmn 768 0267 1350 plusmn 801 1774 plusmn 1517 0358

Values are shown as mean plusmn standard deviation or number (percentage) CVD cardiovascular disease FH family history DM diabetes mellitus BMI bodymass index FBS fasting blood sugar HDL-C high density lipoprotein cholesterol LDL-C low density lipoprotein cholesterol CK total creatine kinase CK-MB creatine kinase isoenzyme MB fraction LDH lactate dehydrogenase Two-sided chi-square and Mann-Whitney 119880 tests were usedlowastStatistically significant at 119875 lt 005

in the distribution frequencies of haplotype combinationsbetween MI patients and healthy controls Homozygotesfor Gly

16Gln27

haplotype showed genetic susceptibility forMI in our population samples whereas heterozygote indi-viduals with other haplotype combinations (Gly

16Gln27

+Arg16Gln27

or Gly16Gln27

+ Gly16Glu27) did not show

a significant association with disease risk thus highlight-ing a clear dose-response relationship between Gly

16Gln27

and MI Moreover carriers of Gly16Glu27

haplotype hadlower prevalence of MI disease and showed protectionagainst the occurrence of the disease On the other handArg16Gln27

haplotype did not influence the susceptibilityfor developing MI in our Egyptian population Similarly twoclinical studies showed differential distribution of haplotypefrequencies ofADRB2 gene betweenMI cases and controls inthe American population with evidence showing dominantprotective effect of the Gly

16Glu27

haplotype on MI andother age-related phenotypes whereas the presence of anydetrimental effect was largely associated with the Gln

27allele

and Gly16Gln27

haplotype [71 85] This was in line withother previous studies which reported significant associationbetween Gly

16Gln27

haplotype and low exercise perfor-mance in heart failure patients [82] Consistent with thishypothesis Gly

16Glu27was shown to be the optimal haplo-

type combination for cardiovascular response during exercisebecause of increased receptor numbers and enhanced strokevolume and cardiac output [84 86] and to elicit more 120573-blocker drug response in postischemic heart [87] In contrast

two studies reported a decreased frequency of Gly16Gln27

haplotype in male and female Caucasian patients with MI[13 88] Other studies however did not find associationbetween theADRB2 haplotype and the risk ofMI [25 89 90]We propose two explanations underlying the association ofGly16Gln27

haplotype homozygosity with MI developmentSeveral lines of evidence indicated that ADRB2 like otherG-protein-coupled receptors form dimers The formationof 1205732-AR dimers was shown to have functional effects on

receptor stimulated adenylate cyclase activity [91] So onepossible mechanism is the interaction between two receptorswith Gly

16Gln27haplotype forming ldquohomodimerrdquo may have

different agonist binding signal transduction or agonist-promoted desensitization properties than other types ofdimers However there is an alternative possibility which isthat Arg16Gly and Gln27Glu polymorphisms might be inLD with other polymorphic loci in ADRB2 gene or nearbyinflammatory mediator genes which have direct effect onthe pathogenesis of MI [92] Drysdale et al described othermolecular haplotypic structure ofADRB2 including our SNPs[66] One of them Arg19Cys polymorphism in the 51015840-leadercistron was found to be in LDwith Gln27Glu polymorphism[93] and caused altered receptor translation hence affecting1205732-AR protein expression in vitro [32 94 95]Another main finding in our study was the difference

in genotype distribution of the ADRB2 Gln27Glu polymor-phism between hypertensive and normotensive MI patientsHomozygotes of the Gln

27variant had a significant higher


frequency of hypertension compared to other genotype car-riers This was in agreement with previous studies conductedin Americans European and Han Chinese populations [1192 95] Binder et al reported association between both Gln

27

haplotypes (Gly16Gln27and Arg

16Gln27) and high diastolic

blood pressure in Caucasian males [96] Subjects with theGln27

variant were shown to have attenuated vasodilatoryresponse to infused isoproterenol while those with Glu

27

variant had more robust reversal of constriction than didthe Gln

27in normotensive males [87 97] Furthermore

carriers of the allele Glu27and Gly

16Glu27haplotype showed

increased forearm vasodilatation during mental stress andhandgrip exercise [98] and might cause arterial hypotensionwith thoracic epidural anesthesia [99] presumably due to itsminimal downregulation by catecholamines hence increas-ing 1205732-AR signaling [87] In contrast others demonstrated

that the variation within codons 16 and 27 of ADRB2 genewere unlikely to confer genetic susceptibility for hypertensionin Americans black South Africans and Asians suggestingLD with other unidentified genetic variants [48 56 64 100ndash102]

Assessment of other cardiovascular risk factors in ourMI patients revealed a significant difference in genotypefrequencies of ADRB2 Arg16Gly polymorphism betweendiabetic and nondiabetic MI patients The frequency ofdiabetes mellitus (DM) was significantly higher in patientswith Gly

16Gly16genotype than other genotypes The 120573

2-AR

are known to be expressed in a variety of tissues includingpancreatic beta-cells and to play a key role in the regulation ofglucosemetabolism aswell as insulin sensitivity and secretion[103 104] Masuo et al have reported significant associationbetween the Gly

16variant of the ADRB2 gene with increased

insulin resistance [105] Prior et al suggested that ADRB2haplotypes mediate insulin action glucose tolerance andpotentially risk for type 2 diabetes mellitus in obese post-menopausal women [104] On the contrary homozygosity ofArg16allele in the ADRB2 gene was associated with a higher

frequency for development of type 2 diabetes in Taiwaneseand Danish populations [106 107] while no associationwith type 2 diabetes was found in Koreans [108] Tonganpopulation [109] and young Danish males [110]

In our MI patients Arg16Gln27

haplotype had highertotal cholesterol levels compared to their counterpart Earlierstudies have suggested that the Arg16Gly polymorphismmaybe associated with cholesterol metabolism in certain popula-tions [39] In Han Chinese population Arg

16homozygotes

had higher serum total cholesterol triglycerides and low-density lipoprotein cholesterol levels [111] An inverse asso-ciation was found in Saudian population with higher levels oftotal cholesterol triglycerides and LDL-C inArg

16Gly16and

Gly16Gly16individuals [16] In addition the Gly

16homozy-

gotes had a lower HDL-C level than the Arg16

homozygotesin Japanese population [27] while no association with dys-lipidemia was found with ADRB2 gene polymorphisms atcodons 16 and 27 in Korean population [108]

The inconsistent associations of ADRB2 variants in ourresults with some prior studies could be caused by the geneticheterogeneity among different ethnic groups [59 66 112]since the susceptibility to MI is affected by multiple genetic

factors and genotype-environment interactions (multifacto-rial inheritance mode) in which each factor has a smallinfluence on the development of the disease [4 113] Thusvariations in the frequency of SNPs in different populationsthe modulating effects of other SNPs or mutations withinindividuals variation in the penetrance of a SNP due toother factors as age-effect different gender of enrolled studypopulations and the difficulty in matching for all knownenvironmental factors that predispose to MI could pro-vide other plausible explanations for the conflicting resultsbetween studies Overall our study had few limitations oursample size may be considered small there was no control forchronic pharmacological treatment which may interfere withour findings and only two polymorphisms were analyzed Inaddition the Egyptian population is admixed and a regionwith a specific genotype combination associated with riskmay also be associated with a peculiar environmental factor

5 Conclusion

In conclusion the present study provides the first preliminaryevidence that Gln27Glu polymorphism of 120573

2-adrenergic

receptors but not the Arg16Gly polymorphism is relatedto the prevalence of myocardial infarction disease in Egyp-tian population Identification of the molecular mechanismsunderlying the relationship between 120573

2-adrenergic receptor

genes andmyocardial infarction offers opportunities to iden-tify individuals at high risk and may help to improve theprevention and treatment of this important disease

Conflict of Interests

The authors declare that the research was conducted in theabsence of any commercial or financial relationships thatcould be considered as a potential conflict of interests

Authorsrsquo Contribution

Eman Toraih Mohammad H Hussein and Dahlia I Badranconceived and designed the experiments Dahlia I Badrancollected samples and data Eman Toraih Mohammad HHussein and Dahlia I Badran performed the experimentsEman Toraih and Dahlia I Badran analyzed the dataEman Toraih Mohammad H Hussein and Dahlia I Badrancontributed reagentsmaterialsanalysis tools Eman ToraihMohammad H Hussein and Dahlia I Badran wrote thepaper Eman Toraih Mohammad H Hussein and DahliaI Badran were responsible for critical revision and finalapproval

Acknowledgment

The authors thank Dr Dalia Sabry the Lab Manager atBiotechnology Research Center for her technical support

References

[1] Q Wang S-B Zhou and L-J Wang ldquoSeven functional poly-morphisms in the CETP Gene and myocardial infarction risk


a meta-analysis and meta-regressionrdquo PLoS One vol 9 no 2Article ID e88118 2014

[2] KThygesen and J Searle ldquoUpdate on the universal definition ofacutemyocardial infarction in the light of new datardquoConferencePapers in Medicine vol 2013 Article ID 479352 5 pages 2013

[3] C de Zwaan M Daemen and W Hermens ldquoMechanisms ofcell death in acute myocardial infarction pathophysiologicalimplications for treatmentrdquoNetherlandHeart Journal vol 9 no1 pp 30ndash44 2001

[4] D H Walter and A M Zeiher ldquoGenetic risk factors formyocardial infarctionrdquo Herz vol 25 no 1 pp 7ndash14 2000

[5] Y Yamada H Izawa S Ichihara et al ldquoPrediction of the riskof myocardial infarction from polymorphisms in candidategenesrdquo The New England Journal of Medicine vol 347 no 24pp 1916ndash1923 2002

[6] S Visvikis-Siest and J-B Marteau ldquoGenetic variants predispos-ing to cardiovascular diseaserdquo Current Opinion in Lipidologyvol 17 no 2 pp 139ndash151 2006

[7] B Cao Y Z Ye J Rui et al ldquoA single-nucleotide polymorphismin the proximal promoter region of the apolipoproteinMgene isassociatedwith dyslipidaemia but not increased coronary arterydiseases in Chinese populationsrdquo Lipids in Health and Diseasevol 12 no 1 article 184 2013

[8] G Abraham O G Bhalala P I W de Bakker S Ripatti andMInouye ldquoTowards amolecular systemsmodel of coronary arterydiseaserdquo Current Cardiology Reports vol 16 no 6 article 4882014

[9] E J Topol J Smith E F Plow andQKWang ldquoGenetic suscep-tibility to myocardial infarction and coronary artery diseaserdquoHuman Molecular Genetics vol 15 no 2 pp R117ndashR123 2006

[10] H Schunkert J Erdmann and N J Samani ldquoGenetics ofmyocardial infarction a progress reportrdquo European Heart Jour-nal vol 31 no 8 pp 918ndash925 2010

[11] L A Hindorff S R Heckbert B M Psaty et al ldquo1205732-adrenergicreceptor polymorphisms and determinants of cardiovascularrisk the Cardiovascular Health StudyrdquoThe American Journal ofHypertension vol 18 no 3 pp 392ndash397 2005

[12] E M Snyder M J Joyner S T Turner and B D JohnsonldquoBlood pressure variation in healthy humans a possible interac-tion with 120573-2 adrenergic receptor genotype and renal epithelialsodium channelsrdquo Medical Hypotheses vol 65 no 2 pp 296ndash299 2005

[13] R Y L ZeeN RCook R Reynolds S Cheng andPMRidkerldquoHaplotype analysis of the 1205732 adrenergic receptor gene and riskofmyocardial infarction in humansrdquoGenetics vol 169 no 3 pp1583ndash1587 2005

[14] A Yilmaz M G Kaya U Merdanoglu M A Ergun A Cengeland SMenevse ldquoAssociation of 120573-1 and 120573-2 adrenergic receptorgene polymorphisms with myocardial infarctionrdquo Journal ofClinical Laboratory Analysis vol 23 no 4 pp 237ndash243 2009

[15] A M Kulminski I V Culminskaya S V Ukraintseva et alldquoPolymorphisms in the ACE and ADRB2 genes and risks ofaging-associated phenotypes the case ofmyocardial infarctionrdquoRejuvenation Research vol 13 no 1 pp 13ndash21 2010

[16] M H Daghestani A Warsy A N Al-Odaib et al ldquoArginine 16glycine polymorphism in 1205732-adrenergic receptor gene is associ-ated with obesity hyperlipidemia hyperleptinemia and insulinresistance in saudisrdquo International Journal of Endocrinology vol2012 Article ID 945608 8 pages 2012

[17] S Guimaraes and D Moura ldquoVascular adrenoceptors anupdaterdquo Pharmacological Reviews vol 53 no 2 pp 319ndash3562001

[18] H A Rockman W J Koch and R J Lefkowitz ldquoSeven-trans-membrane-spanning receptors and heart functionrdquoNature vol415 no 6868 pp 206ndash212 2002

[19] S RHeckbert L AHindorff K L Edwards et al ldquo1205732-adrener-gic receptor polymorphisms and risk of incident cardiovascularevents in the elderlyrdquo Circulation vol 107 no 15 pp 2021ndash20242003

[20] M Zaugg M C Schaub T Pasch and D R Spahn ldquoModula-tion of 120573-adrenergic receptor subtype activities in perioperativemedicine mechanisms and sites of actionrdquo British Journal ofAnaesthesia vol 88 no 1 pp 101ndash123 2002

[21] J A Johnson and S B Liggett ldquoCardiovascular pharmacoge-nomics of adrenergic receptor signaling clinical implicationsand future directionsrdquo Clinical Pharmacology andTherapeuticsvol 89 no 3 pp 366ndash378 2011

[22] J A Johnson and S G Terra ldquo120573-adrenergic receptor polymor-phisms cardiovascular disease associations and pharmacoge-neticsrdquo Pharmaceutical Research vol 19 no 12 pp 1779ndash17872002

[23] A Al-Rubaish F Al-Muhanna A Al-Shehri et al ldquo1205732-adrener-

gic receptor gene polymorphisms in normal and in patientswithmyocardial infarction in the eastern province of Saudi ArabiardquoSaudi Journal ofMedicine andMedical Sciences vol 1 pp 25ndash292013

[24] L P Svetkey P Z Timmons O Emovon N B Anderson LPreis and Y-T Chen ldquoAssociation of hypertension with beta2-and alpha2c10-adrenergic receptor genotyperdquo Hypertensionvol 27 no 6 pp 1210ndash1215 1996

[25] S-M Herrmann V Nicaud L Tiret et al ldquoPolymorphisms ofthe 1205732-adrenoceptor (ADRB2) gene and essential hypertensionthe ECTIM and PEGASE studiesrdquo Journal of Hypertension vol20 no 2 pp 229ndash235 2002

[26] F Galletti R Iacone E Ragone et al ldquoLack of associationbetween polymorphism in the 1205732-adrenergic receptor genehypertension and obesity in the Olivetti Heart Studyrdquo TheAmerican Journal of Hypertension vol 17 no 8 pp 718ndash7202004

[27] Y Iwamoto M Ohishi M Yuan et al ldquo120573-Adrenergic receptorgene polymorphism is a genetic risk factor for cardiovasculardisease a cohort study with hypertensive patientsrdquo Hyperten-sion Research vol 34 no 5 pp 573ndash577 2011

[28] C Forleo N Resta S Sorrentino et al ldquoAssociation of 120573-adrenergic receptor polymorphisms and progression to heartfailure in patients with idiopathic dilated cardiomyopathyrdquoTheAmerican Journal of Medicine vol 117 no 7 pp 451ndash458 2004

[29] S B Liggett L E Wagoner L L Craft et al ldquoThe Ile1641205732-adrenergic receptor polymorphism adversely affects theoutcome of congestive heart failurerdquo The Journal of ClinicalInvestigation vol 102 no 8 pp 1534ndash1539 1998

[30] J Shin M T Lobmeyer Y Gong et al ldquoRelation of beta(2)-adrenoceptor haplotype to risk of death and heart trans-plantation in patients with heart failurerdquo American Journal ofCardiology vol 99 no 2 pp 250ndash255 2007

[31] C Ulucan V Cetintas A Tetik et al ldquo1205731 and 1205732-adrenergic

receptor polymorphisms and idiopathic ventricular arrhyth-miasrdquo Journal of Cardiovascular Electrophysiology vol 19 no10 pp 1053ndash1058 2008

[32] N Sotoodehnia D S Siscovick M Vatta et al ldquo1205732-adrenergicreceptor genetic variants and risk of sudden cardiac deathrdquoCirculation vol 113 no 15 pp 1842ndash1848 2006


[33] L P ChungGWaterer andP JThompson ldquoPharmacogeneticsof 1205732 adrenergic receptor gene polymorphisms long-acting 120573-agonists and asthmardquo Clinical and Experimental Allergy vol 41no 3 pp 312ndash326 2011

[34] R A Karam N A Sabbah H E Zidan and H M A RahmanldquoAssociation between genetic polymorphisms of 1205732 adrenergicreceptors and nocturnal asthma in Egyptian childrenrdquo Journalof Investigational Allergology and Clinical Immunology vol 23no 4 pp 262ndash266 2013

[35] A C Z de Paiva F A D L Marson J D Ribeiro and CS Bertuzzo ldquoAsthma gln27Glu and Arg16Gly polymorphismsof the beta2-adrenergic receptor gene as risk factorsrdquo AllergyAsthma and Clinical Immunology vol 10 no 1 article 8 2014

[36] C Hesse and J H Eisenach ldquoGenetic variation in the 1205732-

adrenergic receptor impact on intermediate cardiovascu-lar phenotypesrdquo Current Pharmacogenomics and PersonalizedMedicine vol 6 no 3 pp 160ndash170 2008

[37] A M Al-Rubaish ldquoAssociation of beta2-adrenergic receptor

gene polymorphisms and nocturnal asthma in Saudi patientsrdquoAnnals of Thoracic Medicine vol 6 no 2 pp 66ndash69 2011

[38] M Johnson ldquoMolecular mechanisms of 1205732-adrenergic receptorfunction response and regulationrdquo Journal of Allergy andClinical Immunology vol 117 no 1 pp 18ndash24 2006

[39] A A Litonjua L Gong Q L Duan et al ldquoVery important phar-macogene summary ADRB2rdquo Pharmacogenetics and Genomicsvol 20 no 1 pp 64ndash69 2010

[40] S B Pereira I A Gava C Giro and E TMesquita ldquoAdrenergicreceptor polymorphisms in heart failure what can geneticsexplainrdquo Arquivos Brasileiros de Cardiologia vol 94 no 6 pp841ndash849 2010

[41] K Masuo and G W Lambert ldquoRelationships of adrenoceptorpolymorphisms with obesityrdquo Journal of Obesity vol 2011Article ID 609485 10 pages 2011

[42] K S Adriani M C Brouwer F Baas A H Zwinderman Avan der Ende and D van de Beek ldquoGenetic variation in the 1205732-adrenocepter gene is associated with susceptibility to bacterialmeningitis in adultsrdquo PLoS ONE vol 7 no 5 Article ID e376182012

[43] O E Brodde ldquo120573-1 and 120573-2 adrenoceptor polymorphismsfunctional importance impact on cardiovascular diseases anddrug responsesrdquo Pharmacology and Therapeutics vol 117 no 1pp 1ndash29 2008

[44] G Sala A di Castelnuovo L Cuomo et al ldquoThe E27 1205732-

adrenergic receptor polymorphism reduces the risk of myocar-dial infarction in dyslipidemic young malesrdquo Journal of Throm-bosis and Haemostasis vol 85 pp 231ndash233 2001

[45] S M Wallerstedt A L Eriksson C Ohlsson and T Hed-ner ldquoHaplotype association analysis of the polymorphismsArg16Gly and Gln27Glu of the adrenergic 1205732 receptor in aSwedish hypertensive populationrdquo Journal of Human Hyperten-sion vol 19 no 9 pp 705ndash708 2005

[46] K Thygesen J Alpert and A Jaffe ldquoThird universal definitionof myocardial infarctionrdquo European Heart Journal vol 33 no20 pp 2551ndash2567 2012

[47] American Diabetes Association ldquoDiagnosis and classificationof diabetes mellitusrdquo Diabetes Care vol 33 supplement 1 ppS62ndashS69 2010

[48] Y Lou J Liu Y Li et al ldquoAssociation study of the 1205732-adrenergic receptor gene polymorphisms and hypertension inthe Northern Han Chineserdquo PLoS ONE vol 6 no 4 Article IDe18590 2011

[49] P Trinder ldquoDetermination of blood glucose using an oxidase-peroxidase system with a non-carcinogenic chromogenrdquo Jour-nal of Clinical Pathology vol 22 no 2 pp 158ndash161 1969

[50] C C Allain L Poon and C Chan ldquoEnzymatic determinationof total serum cholesterolrdquo Clinical Chemistry vol 20 no 4 pp470ndash475 1974

[51] B Roschlau E Burnt and W Gruber ldquoEnzymatiche estim-mung des cesamt-cholestrins in serumrdquo Zeitschrift fur klinischeChemie und klinische Biochemie vol 12 p 226 1974

[52] J Siedel R Schmuck and J Staepels ldquoLong-term stable liquidready-to-use mono reagent for the enzymatic assay of serum orplasma triglycerides (GPO-PAP method)rdquo Clinical Chemistryvol 39 AACC meeting abstract 34 p 1127 1993

[53] M F Lopes Virella P Stone S Ellis and J A Colwell ldquoCholes-terol determination in high density lipoproteins separated bythree different methodsrdquo Clinical Chemistry vol 23 no 5 pp882ndash884 1977

[54] K Shashidhar P Pareenta B Gopalakrishna and AHemalatha ldquoLipoprotein (a) and other lipid profile inpatients with thrombotic stroke is it a reliable markerrdquo Journalof Laboratory Physicians vol 3 no 1 pp 28ndash32 2011

[55] J Gennest and P Libby ldquoLipoprotein disorders and cardio-vascular diseaserdquo in Braunwaldrsquos Heart Disease A Textbook ofCardiovascular Medicine R O Bonow D L Mann D P Zipesand P Libby Eds chapter 47 Saunders Elsevier PhiladelphiaPa USA 9th edition 2011

[56] J Jung P Min H Park et al ldquoThe effects of the 1205732-adrenergic

receptor gene polymorphism on the risk of essential hyperten-sionrdquo Korean Circulation Journal vol 35 no 10 pp 753ndash7582005

[57] A Blakemore C Angela G Ana-Maria et al ldquoIL-Ra allele(IL-1RNlowast2) associated with nephropathy in diabetesrdquo HumanGenetics vol 97 no 3 pp 369ndash374 1996

[58] J Syrjanen M Hurme T Lehtimaki J Mustonen and APasternack ldquoPolymorphism of the cytokine genes and IgAnephropathyrdquoKidney International vol 61 no 3 pp 1079ndash10852002

[59] A E Hegab T Sakamoto W Saitoh et al ldquoPolymorphisms ofIL4 IL13 andADRB2 genes in COPDrdquoChest vol 126 no 6 pp1832ndash1839 2004

[60] D G Contopoulos-Ioannidis E N Manoli and J P AIoannidis ldquoMeta-analysis of the association of 1205732-adrenergicreceptor polymorphisms with asthma phenotypesrdquo Journal ofAllergy and Clinical Immunology vol 115 no 5 pp 963ndash9722005

[61] K K Abu-Amero O M Al-Boudari G H Mohamed andN Dzimiri ldquoThe Glu27 genotypes of the Beta2-adrenergicreceptor are predictors for severe coronary artery diseaserdquo BMCMedical Genetics vol 7 article 31 2006

[62] H G Xie C M Stein R B Kim et al ldquoFrequency of func-tionally important beta-2 adrenoceptor polymorphisms variesmarkedly among African-American Caucasian and Chineseindividualsrdquo Pharmacogenetics vol 9 no 4 pp 511ndash516 1999

[63] T Limsuwan A Thakkinstian O Verasertniyom et al ldquoPos-sible protective effects of the Glu27 allele of 1205732-adrenergicreceptor polymorphism in Thai asthmatic patientsrdquo AsianPacific Journal of Allergy and Immunology vol 28 no 2-3 pp107ndash114 2010

[64] A E Atia K Norsidah A Nor Zamzila et al ldquoPreliminarystudy on association of 120573

2-adrenergic receptor polymorphism

with hypertension in hypertensive subjects attending Balok


Health Centre Kuantanrdquo Medical Journal of Malaysia vol 67no 1 pp 25ndash30 2012

[65] M DrsquoAmato L R Vitiani G Petrelli et al ldquoAssociation ofpersistent bronchial hyperresponsiveness with120573

2-adrenoceptor

(ADRB2) haplotypes a population studyrdquo American Journal ofRespiratory and Critical Care Medicine vol 158 no 6 pp 1968ndash1973 1998

[66] C M Drysdale D W McGraw C B Stack et al ldquoComplexpromoter and coding region 120573

2-adrenergic receptor haplotypes

alter receptor expression and predict in vivo responsivenessrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 97 no 19 pp 10483ndash10488 2000

[67] E Israel V M Chinchilli J G Ford et al ldquoUse of regularlyscheduled albuterol treatment in asthma genotype-stratifiedrandomised placebo-controlled cross-over trialrdquo The Lancetvol 364 no 9444 pp 1505ndash1512 2004

[68] G A Hawkins K Tantisira D A Meyers et al ldquoSequencehaplotype and association analysis of ADR1205732 in a multiethnicasthma case-control studyrdquoAmerican Journal of Respiratory andCritical Care Medicine vol 174 no 10 pp 1101ndash1109 2006

[69] G Vacca K Schwabe R Duck et al ldquoPolymorphisms of the 1205732

adrenoreceptor gene in chronic obstructive pulmonary diseaserdquoTherapeutic Advances in Respiratory Disease vol 3 no 1 pp 3ndash10 2009

[70] K Leineweber and G Heusch ldquo1205731- and 120573

2-adrenoceptor

polymorphisms and cardiovascular diseasesrdquo British Journal ofPharmacology vol 158 no 1 pp 61ndash69 2009

[71] A M Kulminski I Culminskaya S V Ukraintseva K GArbeev K C Land andA I Yashin ldquoBeta2-adrenergic receptorgene polymorphisms as systemic determinants of healthy agingin an evolutionary contextrdquoMechanisms of Ageing and Develop-ment vol 131 no 5 pp 338ndash345 2010

[72] N Pezzali A Curnis C Specchia et al ldquoAdrenergic receptorgene polymorphism and left ventricular reverse remodellingafter cardiac resynchronization therapy preliminary resultsrdquoEuropace vol 15 no 10 pp 1475ndash1481 2013

[73] D M Kaye B Smirk C Williams G Jennings M Esler andD Holst ldquo120573-adrenoceptor genotype influences the response tocarvedilol in patients with congestive heart failurerdquo Pharmaco-genetics vol 13 no 7 pp 379ndash382 2003

[74] E Barbato A Berger L Delrue et al ldquoGLU-27 variant of1205732-adrenergic receptor polymorphisms is an independent riskfactor for coronary atherosclerotic diseaserdquo Atherosclerosis vol194 no 2 pp e80ndashe86 2007

[75] J Li Y Zhang C Li et al ldquoHSPA12B attenuates cardiac dys-function and remodelling after myocardial infarction throughan eNOS-dependentmechanismrdquoCardiovascular Research vol99 no 4 pp 674ndash684 2013

[76] M Johnson ldquoThe 120573-adrenoceptorrdquo American Journal of Respi-ratory and Critical Care Medicine vol 158 no 5 pp S146ndashS1531998

[77] S A Green J Turki P Bejarano I P Hall and S B LiggettldquoInfluence of beta 2-adrenergic receptor genotypes on signaltransduction in human airway smooth muscle cellsrdquo Americanjournal of respiratory cell andmolecular biology vol 13 no 1 pp25ndash33 1995

[78] S Green J Turki and M Innis ldquoAmino-terminal polymor-phisms of the human 120573

2-adrenergic receptor impart distinct

agonist-promoted regulatory propertiesrdquo Biochemistry vol 33pp 9414ndash9419 1994

[79] K M Small D W McGraw and S B Liggett ldquoPharmacologyand physiology of human adrenergic receptor polymorphismsrdquo

Annual Review of Pharmacology and Toxicology vol 43 pp 381ndash411 2003

[80] S A Green G Cole M Jacinto M Innis and S B LiggettldquoA polymorphism of the human 120573

2-adrenergic receptor within

the fourth transmembrane domain alters ligand binding andfunctional properties of the receptorrdquo The Journal of BiologicalChemistry vol 268 no 31 pp 23116ndash23121 1993

[81] S B Liggett ldquo1205732-Adrenergic receptor pharmacogeneticsrdquo

American Journal of Respiratory and Critical Care Medicine vol161 no 3 pp S197ndashS201 2000

[82] L EWagoner L L Craft B Singh et al ldquoPolymorphisms of the1205732-adrenergic receptor determine exercise capacity in patients

with heart failurerdquo Circulation Research vol 86 no 8 pp 834ndash840 2000

[83] G Iaccarino R Izzo V Trimarco et al ldquo1205732-Adrenergic receptor

polymorphisms and treatment-induced regression of left ven-tricular hypertrophy in hypertensionrdquo Clinical PharmacologyandTherapeutics vol 80 no 6 pp 633ndash645 2006

[84] V Sarpeshkar and D J Bentley ldquoAdrenergic-1205732receptor

polymorphism and athletic performancerdquo Journal of HumanGenetics vol 55 no 8 pp 479ndash485 2010

[85] A Thakkinstian M McEvoy C Minelli et al ldquoSystematicreview and meta-analysis of the association between 120573

2-

adrenoceptor polymorphisms and asthma a HuGE reviewrdquoAmerican Journal of Epidemiology vol 162 no 3 pp 201ndash2112005

[86] V Dishy G G Sofowora H-G Xie et al ldquoThe effect ofcommon polymorphisms of the 120573

2-adrenergic receptor on

agonist-mediated vascular desensitizationrdquo The New EnglandJournal of Medicine vol 345 no 14 pp 1030ndash1035 2001

[87] G W Dorn II ldquoAdrenergic signaling polymorphisms and theirimpact on cardiovascular diseaserdquo Physiological Reviews vol90 no 3 pp 1013ndash1062 2010

[88] M Schurks T Kurth P M Ridker J E Buring and R Y LZee ldquoAssociation between polymorphisms in the 120573

2-adrenergic

receptor gene with myocardial infarction and ischaemic strokein womenrdquoThrombosis and Haemostasis vol 101 no 2 pp 351ndash358 2009

[89] S M Wallerstedt A-L Eriksson C Ohlsson and T Hed-ner ldquoHaplotype association analysis of the polymorphismsArg16Gly and Gln27Glu of the adrenergic 120573

2receptor in a

Swedish hypertensive populationrdquo Journal of Human Hyperten-sion vol 19 no 9 pp 705ndash708 2005

[90] K M Rexrode P M Ridker H H Hegener J E Buring J EManson and R Y L Zee ldquoGenetic variation of the androgenreceptor and risk of myocardial infarction and ischemic strokein womenrdquo Stroke vol 39 no 5 pp 1590ndash1592 2008

[91] T E Hebert S Moffett J-P Morello et al ldquoA peptide derivedfrom a 120573

2-adrenergic receptor transmembrane domain inhibits

both receptor dimerization and activationrdquo The Journal ofBiological Chemistry vol 271 no 27 pp 16384ndash16392 1996

[92] D Ge J Huang J He et al ldquo1205732-adrenergic receptor gene

variations associated with stage-2 hypertension in northernHan Chineserdquo Annals of Human Genetics vol 69 no 1 pp 36ndash44 2005

[93] D W McGraw S L Forbes L A Kramer and S B LiggettldquoPolymorphisms of the 51015840 leader cistron of the human beta2-adrenergic receptor regulate receptor expressionrdquo Journal ofClinical Investigation vol 102 no 11 pp 1927ndash1932 1998

[94] A L Parola and B K Kobilka ldquoThe peptide product of a 51015840leader cistron in the 120573

2adrenergic receptor mRNA inhibits


receptor synthesisrdquoThe Journal of Biological Chemistry vol 269no 6 pp 4497ndash4505 1994

[95] K Bengtsson M Orho-Melander O Melander et al ldquo1205732-

Adrenergic receptor gene variation andhypertension in subjectswith type 2 diabetesrdquoHypertension vol 37 no 5 pp 1303ndash13082001

[96] A Binder E Garcia CWallace et al ldquoHaplotypes of the beta-2adrenergic receptor associate with high diastolic blood pressurein the Caerphilly prospective studyrdquo Journal of Hypertensionvol 24 no 3 pp 471ndash477 2006

[97] J R Cockcroft A G Gazis D J Cross et al ldquo1205732-Adrenoceptor

polymorphism determines vascular reactivity in humansrdquoHypertension vol 36 no 3 pp 371ndash375 2000

[98] I C Trombetta L T Batalha M U P B Rondon et al ldquoGly16+Glu27 1205732-adrenoceptor polymorphisms cause increased fore-arm blood flow responses to mental stress and handgrip inhumansrdquo Journal of Applied Physiology vol 98 no 3 pp 787ndash794 2005

[99] U H Frey J Karlik F Herbstreit and J Peters ldquo1205732-

adrenoceptor gene variants affect vasopressor requirements inpatients after thoracic epidural anaesthesiardquo British Journal ofAnaesthesia vol 112 no 3 pp 477ndash484 2014

[100] G Candy N Samani G Norton et al ldquoAssociation analysis of1205732adrenoceptor polymorphisms with hypertension in a Black

African populationrdquo Journal of Hypertension vol 18 no 2 pp167ndash172 2000

[101] H-G Xie C M Stein R B Kim et al ldquoHuman 1205732-adrenergic

receptor polymorphisms no association with essential hyper-tension in black or white Americansrdquo Clinical Pharmacology ampTherapeutics vol 67 no 6 pp 670ndash675 2000

[102] K Ranade M S Chang C T Ting et al ldquoHigh-throughputgenotyping with single nucleotide polymorphismsrdquo GenomeResearch vol 11 no 7 pp 1262ndash1268 2001

[103] S Ishiyama-Shigemoto K Yamada X Yuan F Ichikawa andKNonaka ldquoAssociation of polymorphisms in the120573

2-adrenergic

receptor gene with obesity hypertriglyceridaemia and diabetesmellitusrdquo Diabetologia vol 42 no 1 pp 98ndash101 1999

[104] S J Prior A P Goldberg and A S Ryan ldquoADRB2 haplotypeis associated with glucose tolerance and insulin sensitivity inobese postmenopausal womenrdquo Obesity vol 19 no 2 pp 396ndash401 2011

[105] K Masuo T Katsuya Y Fu H Rakugi T Ogihara and ML Tuck ldquo120573

2-adrenoceptor polymorphisms relate to insulin

resistance and sympathetic overactivity as early markers ofmetabolic disease in nonobese normotensive individualsrdquoAmerican Journal of Hypertension vol 18 no 7 pp 1009ndash10142005

[106] T J Chang M H Tsai Y D Jiang et al ldquoThe Arg16Glypolymorphism of human 1205732-adrenoreceptor is associated withtype 2 diabetes in Taiwanese peoplerdquo Clinical Endocrinologyvol 57 no 5 pp 685ndash690 2002

[107] A P Gjesing G Andersen K S Burgdorf et al ldquoStudies ofthe associations between functional 120573

2- adrenergic receptor

variants and obesity hypertension and type 2 diabetes in 7808white subjectsrdquo Diabetologia vol 50 no 3 pp 563ndash568 2007

[108] S-H Kim D-J Kim I A Seo et al ldquoSignificance of 1205732-

adrenergic receptor gene polymorphism in obesity and type 2diabetes mellitus in Korean subjectsrdquo Metabolism Clinical andExperimental vol 51 no 7 pp 833ndash837 2002

[109] N L Duarte S Colagiuri T Palu X L Wang and D E LWilcken ldquoObesity Type II diabetes and the 1205732 adrenoceptor

gene Gln27Glu polymorphism in the Tongan populationrdquoClinical Science vol 104 no 3 pp 211ndash215 2003

[110] S M Echwald T I A Soslashrensen A Tybjaeligrg-Hansen T Ander-sen and O Pedersen ldquoGln27Glu variant of the human 120573

2-

adrenoreceptor gene is not associated with early-onset obesityin Danish menrdquo Diabetes vol 47 no 10 pp 1657ndash1658 1998

[111] H Wu H Bai P Fan R Liu Y Liu and B Liu ldquoAnalysis of1205732-adrenergic receptor gene (120573

2-AR) Arg16Gly polymorphism

in patients with endogenous hypertriglyceridemia in Chinesepopulationrdquo Zhonghua Yi Xue Yi Chuan Xue Za Zhi vol 25 pp50ndash54 2008

[112] D Vercelli ldquoDiscovering susceptibility genes for asthma andallergyrdquo Nature Reviews Immunology vol 8 no 3 pp 169ndash1822008

[113] A Kimura ldquoGenetic analysis of multifactorial cardiac diseasesidiopathic dilated cardiomyopathy and myocardial infarctionrdquoNippon Ika Daigaku zasshi vol 66 no 5 pp 332ndash335 1999

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Enzyme Research



Microbiology


many populations [33 34] The most important two SNPsare located within the amino-terminal extracellular domainnear the receptorrsquos ligand-binding site [35] The first SNPwas caused by a change of adenine to guanine (A gt G)at nucleotide 46 resulting in amino acid substitution fromarginine to glycine at codon 16 (Arg16Gly or R16G) Thesecond SNP is the change of cytosine to guanine (C gt G)at nucleotide 79 causing the substitution from glutamineto glutamic acid at codon 27 (Gln27Glu or Q27E) [15 3637] In vivo and in vitro functional studies suggested thatthese two polymorphisms have altered receptor functionand behavior [38ndash42] Several clinical and pharmacologicalstudies highlighted the important role of these polymor-phisms in the pathogenesis of CVD and their impact on drugresponse [22 30 39 43] However evidence regarding therole of ADRB2 gene polymorphisms in patients with MI isambiguous and inconsistent Significant differences in thegenotype frequencies and haplotype structure ofADRB2 genewere noted in different ethnic populations [13 23 25 44 45]Thus this case-control study was conducted to determine theprevalence of these ADRB2 gene polymorphisms in Egyptianpopulation and to assess their genetic association with thesusceptibility to MI disease

2 Materials and Methods

21 Study Participants The study population was composedof 68 patients with acute myocardial infarction (MI) and 75age-matched healthy controls with no history of cardiovas-cular events hypertension or any other chronic diseasesPatients were recruited from Coronary Care Unit (CCU)Suez Canal University Hospital Ismailia Egypt during theperiod between April 2013 and March 2014 The diagnosisof MI was confirmed by evidence of symptoms in thepresence of either diagnostic elevations of cardiac enzymesor diagnostic changes on electrocardiograms (ECG) [46]Thecardiovascular risk factors were assessed in all participantsTheyunderwent evaluations includingmeasurement of bloodpressure body mass index (BMI) fasting serum lipid andglucose levels cardiac enzymes and ECG Diabetic patientseither had a known history of diabetes mellitus or were diag-nosed according to the American Diabetic Association rec-ommendations [47] Hypertensionwas defined as the averageSBP ge 140mmHg andor the average DBP ge 90mmHgandor self-reported current treatment for hypertension withantihypertensive medication [48] The study was approvedby the Medical Research Ethics Committee of Faculty ofMedicine Suez Canal University Written informed consentwas obtained from all participants

22 Samples Collection and Biochemical Measurements Fast-ing blood samples were collected from both cases andcontrols Blood from patients was collected within 24 hoursof CCU admission Two milliliters (mL) of venous blood waswithdrawn in EDTA tubes to be used for subsequent geneticanalysis Another two milliliters was collected in VacutainerSerum Separator Tubes II (Becton Dickinson Plymouth)to obtain serum Parameters such as fasting blood sugar

(FBS) total cholesterol (TC) triglycerides (TG) high den-sity lipoproteins-C (HDL-C) and low density lipoprotein-C(LDL-C) were analyzed within 2 h after collection Fastingblood sugar was estimated using RocheHitachi analyzer kitsbased on the glucose oxidase and peroxidase method [49]Serum TC and TG were measured by enzymatic colorimetricdetermination method using RocheHitachi cholesterol kit(Cat number 1489232) [50 51] and RocheHitachi triglyc-erides kit (Cat number 1488872) [52] HDL-C was measuredusing RocheHitachiHDL-C kit (Cat number 03030024) andwas estimated by phosphotungstic acid precipitation followedby enzymatic analysis in supernatant fraction [53] LDL-Cwas calculated by the Friedewald formula after consideringits limitations if TG gt 400mgdL LDL-C = TC minus (HDL-C +TG5) [54] FBS lt 126mgdL TC levels lt200mgdL TG lev-els lt150mgdL LDL-C lt 130mgdL andHDL-C gt 60mgdLwere considered normal [55]

23 Genotyping Genomic DNA was extracted from bloodleukocytes using QIAamp DNA Blood Mini kit (QIAgenClinilab Co Egypt Catalog number 51104) according to themanufacturerrsquos instructions [56] Concentration andpurity ofthe extracted DNA were measured by NanoDrop ND-1000(NanoDrop Tech Inc Wilmington DE USA) Genotypingof the ADRB2 polymorphisms (Arg16Gly rs1042713 andGln27Glu rs1042714) was performed by real-time poly-merase chain reaction (RT-PCR) technology PCR was per-formed in a 25-120583L reaction volume containing 125120583L Taq-man Universal PCR Master Mix No AmpErase UNG (2x)20 ng genomic DNA diluted to 1125 120583L with DNase-RNase-free water and 125 120583L 20 times TaqMan SNP Genotyping AssayMix (Applied Biosystems Egypt assay ID C 2084764 20 andC 2084765 20) Internal positive controls and no-templatecontrols (no DNA) were included in each run PCR ampli-fication was carried out in an AB 7500HT instrumentwith the Sequence Detection System (SDS) Software version211 (Applied Biosystems Egypt) according to the followingconditions initial denaturation at 95∘C for 10min followedby 40 cycles of 92∘C for 15 sec and 60∘C for 1min Genotypingwas performed blinded to casecontrol status Ten per cent ofthe randomly selected samples were regenotyped in separateruns to exclude the possibility of false genotype calls with100 concordance rate

24 Statistical Analysis Statistical analysis was carried outusing the Microsoft Excel 2010 and the ldquoStatistical Packagefor the Social Sciences (SPSS) for windowsrdquo software ver-sion 20 Odds ratios (OR) with a 95 confidence inter-val (CI) were calculated Categorical variables were com-pared using the chi-square (1205942) while Mann-Whitney 119880and Kruskal-Wallis tests were used to compare continu-ous variables Due to significant differences in some car-diovascular risk factor variables between cases and con-trol participants binary logistic regression analysis wasperformed that included the ADRB2 polymorphism andpotential confounders (age gender smoking status familyhistory of cardiovascular disease prevalence of obesity andhyperlipidemia) A two-tailed 119875 value of 005 was con-sidered statistically significant The allele frequency within



3 Results


16and Gly

16alleles in


and Glu27





27allele


16Gln27 Gly16Gln27



16Glu27


16Gln27haplotype




16Arg16













4 Discussion



16and Gln



16Gly16

and Arg16Gly16

at codon 16and Gln

27Gln27

and Gln27Glu27


16

and Gln27


16and Gln




16Gln27 29 for Gly

16Gln27 and 41





















Arg16Gln27


Gly16Gln27















16Glu27





27or

Glu27[33 59 60 63 70]



the Glu27


27homozygotes with










2-AR or affect




















16Gln27


16Gln27

+Arg16Gln27

or Gly16Gln27



16Gln27




16Glu27


27allele

and Gly16Gln27


16Gln27














27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



3 Results


16and Gly

16alleles in


and Glu27





27allele


16Gln27 Gly16Gln27



16Glu27


16Gln27haplotype




16Arg16













4 Discussion



16and Gln



16Gly16

and Arg16Gly16

at codon 16and Gln

27Gln27

and Gln27Glu27


16

and Gln27


16and Gln




16Gln27 29 for Gly

16Gln27 and 41





















Arg16Gln27


Gly16Gln27















16Glu27





27or

Glu27[33 59 60 63 70]



the Glu27


27homozygotes with










2-AR or affect




















16Gln27


16Gln27

+Arg16Gln27

or Gly16Gln27



16Gln27




16Glu27


27allele

and Gly16Gln27


16Gln27














27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





















Arg16Gln27


Gly16Gln27















16Glu27





27or

Glu27[33 59 60 63 70]



the Glu27


27homozygotes with










2-AR or affect




















16Gln27


16Gln27

+Arg16Gln27

or Gly16Gln27



16Gln27




16Glu27


27allele

and Gly16Gln27


16Gln27














27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












16Glu27





27or

Glu27[33 59 60 63 70]



the Glu27


27homozygotes with










2-AR or affect




















16Gln27


16Gln27

+Arg16Gln27

or Gly16Gln27



16Gln27




16Glu27


27allele

and Gly16Gln27


16Gln27














27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












16Gln27


16Gln27

+Arg16Gln27

or Gly16Gln27



16Gln27




16Glu27


27allele

and Gly16Gln27


16Gln27














27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



27





27









2-AR







16homozygotes


16Gly16and


16homozy-





5 Conclusion


2-adrenergic








Acknowledgment


References















































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology














































































2-adrenoceptor










2-adrenoceptor


























2-







2-adrenergic



2receptor in a



























2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

















2-adrenergic















2-














Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

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