Arthrosclerosis considered one of the most important causes of morbidity and mortality in industrial and developing countries. The rate of myocardial infarction in some countries is about 2 million annually with 1/4 of them is leading to death. This study was a case-control research, which was carried out as cross-sectional project in two groups, healthy and case subjects. The mean age and standard deviation of patients and control groups were nearly equal (55±10). The ELISA technique was applied to measure the serum antibody level. The sample populations in each group were exactly the same (120 person in each group). The gender distribution in case and control group was 44 female, 76 male and 45 female and 75 male, respectively. The Mean±SD of IgM in case group and control groups were 3.10± 2.54 and 1.54±1.00, respectively. TheMeantSD forlgG incase and control groups were 5.90±3.84 and 3.08±1.95, respectively. The differences, between the mean of IgM and IgG in two groups of cases and control statistically were significant (p = 0.0001). In case group the Meani±SD for IgM for men and woman were 2.98±1.97 and 3.17±2.83, respectively, which this differences statistically, were not significant. In case group the Meant±SD forlg G were 5.14±3.45 and 6.35. The above findings indicated that the average of both IgG and IgM are higher among men compared to women. Due to high prevalence of acute coronary syndrome in Iran, the determination of anticardiolipin antibody (IgG) are applied for suspected acute coronary syndrome patients and further prevention measure should be taken for patient with higher serum anticardiolipin antibody. © 2013 Asian Network for Scientific Information

Ahmadi, A.R.

Behnampour, N.

Joshaghani, H.R.

Mansourian, A.R.

Salehi, A.

Golestan University of Medical Sciences Repository

Pakistan Journal of Biological Sciences 16 (2): 92-96, 2013 ISSN 1028-8880 I DOI: 10.3923/pjbs.2013.92.96 © 2013 Asian Network for Scientific Information The Comparison of Acute Myocardial Infarction Serum Anticardiolipin Antibody with Healthy Subjects, in Gorgan (Northern Iran) 1Ali Reza Ahmadi, 1Hamid Reza Joshaghani, 2Azad Reza Mansourian, 3Aref Salehi, 4Nasser Behnampour 1Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gargan, Iran 2Metabolic Disorder Research Center, Golestan University of Medical Sciences, Gargan, Iran 3Cardiovascular Research Center, Gargan Medical SchoolGolestan University of Medical Sciences, Gargan, Iran 4Hematology and Oncology Research Center, Golestan University of Medical Sciences, Gargan, IranAbstract: Arthrosclerosis considered one of the most important causes of morbidity and mortality in industrial and developing countries. The rate of myocardial infarction in some countries is about 2 million annually with 1 /4 of them is leading to death. This study was a case-control research, which was carried out as cross-sectional project in two groups, healthy and case subjects. The mean age and standard deviation of patients and control groups were nearly equal (55±10). The ELISA technique was applied to measure the serum antibody level. The sample populations in each group were exactly the same (120 person in each group). The gender distribution in case and control group was 44 female, 76 male and 45 female and 75 male, respectively. The Mean±SD of IgM in case group and control groups were 3.10± 2.54 and 1.54±1.00, respectively. The Mean±SD for IgG in case and control groups were 5.90±3.84 and 3.08±1.95, respectively. The differences, between the mean ofigM and IgG in two groups of cases and control statistically were significant (p = 0.0001 ). In case group the Mean±SD for IgM for men and woman were 2.98±1.97 and 3.17±2.83, respectively, which this differences statistically, were not significant. In case group the Mean±SD for IgG were 5.14±3.45 and 6.35. The above findings indicated that the average of both IgG and IgM are higher among men compared to women. Due to high prevalence of acute coronary syndrome in Iran, the determination of anticardiolipin antibody (IgG) are applied for suspected acute coronary syndrome patients and further prevention measure should be taken for patient with higher serum anticardiolipin antibody. Key words: Arthrosclerosis, myocardial infarction, anticardiolipin antibody, northern Iran INTRODUCTION Arthrosclerosis considered being one of the most important causes of morbidity and mortality in industrial and developing countries. The rate of myocardial infarction in U.S. is about 2 million annually with 1 /4 of them is leading to death (Cannon, 2003, El-Sayed et al., 2007). The prevalence of coronary vascular disease in central Iran is about 9.3%. It is reported that 38% of all death in Iran are due to heart diseases. The reasons behind arthrosclerosis are genetically related, acquired risk factors, such as high blood cholesterol. Due to nutritional habitual; high blood pressure, obesity, diabetes, cigarettes smoking, low physical activity, some inflammatory mechanisms can disrupt the regulation of fat within the vain and initiate the arthrosclerosis lesion (Sherer and Shoenfeld, 2002). The recent studies indicate the role plagued by immunological responses in induction of arthrosclerosis. These latest response include the chemical component of CRP, other infectious agents, such as Chlamydia pneumonia and autoantibodies. The various auto-antibodies genes, which can cause and mediate the cardiovascular disease, are including cardiolipin, OXLDL, β2GP1 and HSP (Jara et al., 2006; Su et al., 2006). Antiphospholipids immunoglobulin is examples of such autoantibodies. These autoantibodies can be found in healthy subjects as well but their concentration are low and are found partially is short period of time with the individual blood circulation. The phospholipid autoantibodies of non-autoimmune origin are seen in such disorder as HIV and in some cancers (George et al., 1997; Salehi and Mirzaei, 2007; Jara et al., 2006). The majority of phospholipid antibodies are seen in a one autoimmune antibody named, Antibody Phospholipid Syndrome (APS). The other characteristic of APS are thromboembolic and pregnancy abortions. The APS considered as mediator of autoimmunity and arthrosclerosis (George and Shoenfeld, 1997). The antiphospholipids are integrated with various forms of Correspondence  Author: Ali Reza Ahmadi, Faculty of Health and Para Medical, Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran 92 Pak. J. Biol. Sci., 16 (2): 92-96, 2013cardiovascular  diseases,  such  as  myocardial  infractions. (Sherer and Shoenfeld, 2003; Vaarala et al., 1995). Anti Cardiolipin  Antibody  (ACA)  is  an antibody raised against   cardiolipin   and   in   disease   such   as   APS, autonomous abortion, thrombosis  and lupus. Anticardiolipin    antibody   can   be   divided   into   two categories;   1) class-I:   of Immunoglobulin   M   (IgM). Immunoglobulin G  (IgG), Immunoglobulin A (IgA. 2) Dependent o r  non-dependent t o  β-2-glycoprotein.  This test is requested when the signs of a traumatic accident, such as pain, inflammation in lower organs, respiratory disorder, accompanied with pain are indicated. It is also advised   on repeated   pregnancy   abortion   and   lupus. Considering   the  high   scale  of  acute  coronary   heart diseases and in particular acute myocardial infraction and possible role played by anticardiolipin in acute coronary syndromes (Erkkila et al., 2005, Levine and Jacobs, 2001; Sherer and Shoenfeld, 2003, Wu et al., 1997; Sherer et al., 2001) and particularly due to the absence of a documented para clinical tests and reference into the relation between cardiovascular  disorders  and the  blood  antibody  level, this  present  research  project  was  designed  to  address such queries in this region of northern Iran. MATERIALS AND METHODS Type of research: This study was a case control study which was cross-sectionally carried out in two groups. Due to technical difficulties and the absence of other types of heart diseases, the sample population in the case groups predominantly were from those patients with acute myocardial infarctions. Method of sampling: On the base of statistical parameter the exact number of patient was calculated to be 116 but for sake of security and avoiding any mistake 120 patients were selected. Ethical procedure was applied in this study and all patients were informed on the study procedure and the patients entered in this on their own choice. All the patients were matched on such variables as, age and  gender. Data collection: A questioner was used for every patient, clinical   examination   and   serological    measurement was done by cardiologist and clinical laboratory. The information within the questionnaires, the medical examination   files, laboratory tests, were arranged and entered into SPSS statistical software and presented as a tables and figures. The statistical Kolmogorov-Smirnov test was used to compare the mean of two antibodies of IgM and IgG. Statistical analysis: Initially the serum of two antibodies of IgG, IgM should have been compared;  therefor the normal range and positive results in this study should have been determined before the actual measurement Kolmogorov-Smirnov test was applied for the determination of normal range in this study. Using this statistical equation the following results was designed. Positive results: • >5 GPL mL-1• >15 MPL mL-1Normal range: • ≤15 MPLmL-1• ≤15 GPL mL-1RESULTS The mean age and standard deviation of patients and control groups were nearly equal (55±10).  The sample population in each group was exactly the same (120 person in each group). The gender distribution in case and control groups were 44 female, 76 male and 45 female and 7 5 male, respectively. In this study the matching among case and control was performed properly according to Table 1 and even the gender matching was also were established to obtain the proper results. The Mean±SD of Immunoglobulin M (IgM) in case group and control groups were 3.10±2.54 and 1.54±1.00, respectively. The Mean±SD for Immunoglobulin G IgG in case and control groups were 5.90±3.84 and 3.08±1.95, respectively. The differences, between the mean of IgM and IgG in two groups of cases and control statistically were significant (p 0.0001). In case group the Mean±SD for IgM for men and woman were 2.98±1.97 and 3.1 7±2.83 respectively which this differences statistically were not significant (p 0.697). In case group the Mean±SD for IgG were 5.14±3.45 and 6.35±4.01, respectively. The above findings indicated that the average of both IgG and IgM are higher among men compared to women but this Table 1: The matching among case and control in general in one hand and between two genders of male and females Sex Count Mean±SD p-value p-value (caset+control) Case Female 44 55.29±9.800 0.813 0.99 Male 76 54.84±10.27 Total 55.01±10.06 Control Female 45 55.24±9.700 0.833 Male 75 54.84±10.37 Total 54.99±10.09 Pak. J. Biol. Sci., 16 (2): 92-96, 2013Table 2:  Status of antiphospholipids antibodies of  immunoglobulin G (IgG) and immunoglobulin M  (IgM) among  male and  female  in case and Control groups SexCase Control IgM IgG IgM IgGFemale Mean 2.98 5.14 1.67 3.06 N 44.00 44.00 45.00 45.00 Std.deviation 1.97 3.45 1.23 1.97 Male Mean 3.17 6.35 1.46 3.09 N 76.00 76.00 75.00 75.00 Std.deviation 2.83 4.01 0.84 1.95 Total Mean 3.10 5.90 1.54 3.08 N 120.00 120.00 120.00     120.00 Std.deviation 2.54 3.85 1.00 1.95 Table 3: Serum immunoglobulin G (IgG) and   immunoglobulin   M (IgM) concentration  level  among patient  and healthy subjects lgG IgM Parameters High Normal High Normal Case count Control count  p-value Odds ratio for group (case/control) 95% confidence Interval Values in brackets are percentage 12 (10) 1(0.8) <0.002 13.222 1.691-103.385 108 (90) 119 (99.2) 13 (10.8) 1 (0.8) <0.001 14.458 1.860-112.380 107 (89.2) 119 (99.2) difference  statistically was  not  meaningful (p 0.095) (Table 2). In this research project the serum  IgG  and IgM concentration was determined among  case  and  control groups and the results were tabulated in Table 3 which is shown that there is a statistically significant correlation between patients and healthy subject's antibodies  level.  In control group (Table 2) the Mean±SD of IgM for women and men were 1.67±1.23 and 1.46±0.84, respectively which    statistically   were   not significant (p 0.266).  The Mean±SD for were 3 . 0 4 ±1.95 and 3.06±1.97, which again it was not statistically significant (P+0.917). The comparison of mean of IgM and IgG in both group of case and control among women and men were as follow. Women: The Mean±SD for IgM is both case and control groups were 2.98±1.97 and 1.67±1.23; these findings statistically were significant (p 0.0003). The Mean±SD for IgM in both case and control groups were 5.14±3.45 and 3.06±1.97, respectively; this latter difference statistically were significant (p 0.0007). Men: The Mean±SD of IgM in both case and control group were 3.17±2.83 and 1.46±0.83 which statistically were significant (p 0.00001). The Mean±SD for IgM in both groups of case and control were 6.35±3.09 and 3.09±1.95, respectively. DISCUSSION Acute myocardial infarction and Unstable Angina (UA) are among wide range of recognized diseases, collectively known   as   acute coronary s yndrome.  The pathophysiology of this diseases are due to displace, or disruption of atheroma from coronary internal vascular system. Acute coronary syndromes are among the most common form of sudden death and it is well established that about 50% of AMI patients die before reaching to the hospitals. Earlier studies on the serum Anticardiolipin antibody, level in AMI and its normal range and the relationship of this antibody with AMI and related morbidity and mortality are found to have contradictory, results (Brey et al., 2001; George and Shoenfeld, 1997). Our study indicated that there are a significant different between the serum cardiolipin antibody of AMI patient compared with the control group. In our study we found that there is a significant correlation between the Alv1I and IgG, IgM serum concentration levels. Also in this study we found that Patient/Healthy ratio was found to be 13.22 and 14.458, for IgG and IgM, respectively compared to the subjects which they have normal IgG and IgM serum concentration levels. Anticardiolipin antibody is considered as a major characteristic of antiphospholipids syndrome and in the other hand there is a tendency toward thrombosis onset. It is probable, that anticardiolipin antibody play an important role in occurrence and intensity of AMI due to vascular abnormality. In a study the effective role of anticardiolipin antibody and its relationship with arthrosclerosis in mice it was demonstrated that the mice which were immunized with anticardiolipin antibody and having the higher, titer of anticardiolipin antibody and arthrosclerosis intensity were observed among mice compared to control group (Sherer and Shoenfeld, 2002; George et al., 1997). In agreement   with   our   study    it   is   demonstrated  that Pak. J. Biol. Sci., 16 (2): 92-96, 2013 higher anticardiolipin antibody serum concentration is considered as risk factor, in AMI and in elderly men. (Sherer and Shoenfeld, 2002). In separate study on 63 patients afflicted with the ACS, which were admitted to the CCU during 2003-04, the average anticardiolipin antibody among patient were 4.35±2.7 and in the control group it was 3.38±3.1 which demonstrated to have significant differences (p = 0.038). This average for men in case and control group were 4.47±2.22 and 3.08±2.17, respectively which statistically were significant (p = 0.002). In latter study, there was not a meaningful difference among female in case and control group, 4.1±3.58 vs. 3.69± 3.89 (p = 0.634). The latter finding are in agreement with our general population average and also men average but our finding about women is contradicted other results, because, in our study we found an significant difference among women in case and control group (p = 0.0007). In our study we found also a statistically significant correlation of higher level of IgG and IgM levels among AMI patients which is an interesting finding which can be used in laboratory investigation for AMI patient clinical setting. Also we should mention that in our study we found that there is not meaningful correlation between IgG and IgM levels and AMI among women but there is a meaningful correlation between IgG and IgM serum level and AMI among men, which is an interesting finding . This disagreement can be explained as follow, in Salehi and Mirzaei (2007) study the sample population of case and control were not equal 63 vs. 103. The other explanation is that, in our study in spite of extra clinical and Para clinical examination such as ANA test, we were not able to discriminate the coexistence of other disease such as lupus but we are in opinion that the former possibility can fit these differences. One further clue to this disagreement may be relay on the differences on the women mean age in case and control group. In other study by Ghaemian et al it was demonstrated that there was a statistically significant differences between serum anticardiolipin antibody among patient with AMI and control group (p = 0.01) It was found that there was, a significant increase of serum anticardiolipin antibody of AMI patient with control group (37.14±30.18 vs. 13.14±3.02) and based on their findings it was concluded that anticardiolipin antibody serum level can be considered as a risk factor for ischemic heart diseases and it is suggested it can be considered as marker for the subsequent syndrome vascular thrombolytic lesion (El-Sayed et al., 2007). The latter findings are in agreements with our data from this study, which indicated that the AMI patients have high anticardiolipin antibody serum level, compared with control group but it should mentioned that we measured IgM was measured in addition to IgG which was determined by Elsheikh et al. (2010 ). CONCLUSION Based on our finding we can indicate that the mean serum level of IgM and IgG were 4.55-0.55 and 9.9- 1.15 respectively. The sensitivity and specificity for this spectrum of reference rang was calculated as 90%. It is concluded that due to high prevalence of acute coronary syndrome in Iran particularly among men the determination of anticardiolipin antibody (IgG, IgM) are to be applied for the suspected acute coronary syndrome patients and further prevention measure should be taken for the patients with higher serum anticardiolipin antibody. REFERENCES Brey R.L., RD. Abbott, J.D. Curb, D.S. Sharp, G.W. Ross, C.L. Stallworth and S.J. Kittner, 2001. ~2-glycoprotein I-dependent anticardiolipin antibodies and risk of ischemic stroke and myocardial infarction: The honolulu heart program. Stroke, 32: 1701-1706. Cannon, C.P., 2003. The next step in cardiovascular protection. Atherosclerosis Suppl., 4: 3-9. El-Sayed, M.H., K.M. Ahmed, LAA Shamaa and A.AH. Ghaleb, 2007. 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The comparison of acute myocardial infarction serum anticardiolipin antibody with healthy subjects, in Gorgan (northern Iran)

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