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.v�/�M+P0[P�/%X�.%X�/�/�%X�R04�$��%��/S&|�&��$�$�$�M�M�.0�$�$�$[P|'|'|'|'��������������������������������������������������������������������%X�$�$�$�$�$�$�$�$�$� �:	@Potential link between Helicobacter pylori seropositivity and macrophage migration inhibitory factor in patients with myocardial infarction underwent primary percutaneous coronary intervention 

*Feyan Mirdan Abdullah, MD (master degree)1, Ruqaya Mohammed Ghareeb Taher Al-Barzinji, PhD2
1 Department of Medical Microbiology, Unit of Basic Sciences, College of Medicine, Hawler Medical University, 
  Erbil, Iraq.
2Department of Medical Microbiology, Unit of Basic Sciences, College of Medicine, Hawler Medical University, 
  Erbil, Iraq.
*Corresponding author:  Feyan Mirdan Abdullah
  E-mail:  HYPERLINK "mailto:feyan.mirdan@hmu.edu.krd" feyan.mirdan@hmu.edu.krd

















Abstract 
Objective: To determine the possible link between helicobacter pylori (HP) seropositivity and macrophage migration inhibitory factor (MIF) level in myocardial infarction (MI) patients admitted for primary percutaneous coronary intervention (PPCI).
Method: This case control study was carried out on 100 MI, 38 stable angina pectoris (SAP), and 38 subjects as control who admitted to surgical specialty hospital-cardiac centre in Erbil City, Iraq from January to December 2017. Levels of MIF and anti-HP IgG, IgA, and IgM were detected by enzyme linked immunosorbent assay (ELISA). Polymerase chain reaction (PCR) was done to detect HP 16SrRNA in those samples with IgM+ and/or IgG+IgA+. 
Results: Anti-Hp-IgG was positive in 68%, and 55.26% of MI, and SAP patients respectively.  A higher proportion of HP seropositivity was observed in control group (84.21%). Molecular analysis showed no positive result for HP genomic material. MIF level was insignificantly differed between seropositive and seronegative MI patients (p>0.05). Seropositive MI group had higher MIF level compared to seropositive and negative control groups (p<0.05).
Conclusion: Result of current study suggested that HP does not have direct role in MI elevated MIF, consequently demonstrated secondary contribution of HP in the development and progression of MI through low grade inflammatory response represented by insignificant elevation of MIF, and other inflammatory biomarkers (hsCRP, and fibrinogen).  

Introduction 
The role of chronic infection in atherosclerosis has been investigated in many studies recently, and majority of these studies have examined persistent inflammation resulted from chronic infection with certain microbes ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"abstract":"BACKGROUND: There are conflicting data on the association between Helicobacter pylori (HP) infection and cardiovascular diseases. AIM: To determine if there is an association between gastric HP infection and atherosclerosis of cerebral or peripheral arteries in elderly subjects. METHODS: 90 dyspeptic elderly subjects had upper gastro-intestinal endoscopy and the gastroduodenal pathology was identified. HP infection was confirmed by gastric histology and the rapid urease test. Vascular ultrasonography of extracranial cerebral arteries and leg arteries was performed to evaluate (i) the presence of an atherosclerotic lesion, (ii) the total length of all plaques documented and (iii) the number of arteries with atherosclerotic lesions. Statistical analysis was by the chi2 test, Yates's corrected chi2 test, the Mann-Whitney test and logistic regression. RESULTS: 59 subjects were HP-positive. These had a higher prevalence of peptic ulcer disease (P = 0.01) and higher serum levels of IgG anti-HP antibodies (P = 0.0001), but no significant differences in the number of atherosclerotic lesions, the total length of the plaques or the number of arteries with lesions. No significant association of HP positivity was found with diabetes mellitus, hypertension, cigarette smoking or coronary heart disease, nor with serum concentrations of HDL-cholesterol, fibrinogen, triglycerides or glucose. CONCLUSIONS: Elderly dyspeptic subjects with gastric HP infection had significantly more peptic ulcer disease but no more atherosclerotic lesions than those who were HP-negative. Atherosclerosis was not associated with HP infection. In this cross-sectional study of elderly patients with dyspepsia, no association between HP infection and extracardiac atherosclerosis was found.","author":[{"dropping-particle":"","family":"Pilotto A, Rumo f, Franceschi M, Leandro G, Novello R, Soffiati G, Scagnelli M, Di Mario F","given":"Valerio G","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"age and aging","id":"ITEM-1","issued":{"date-parts":[["1999"]]},"page":"367-371.","title":"Lack of Association between Helicobacter pylori Infection and Extracardiac Atherosclerosis in Dyspepetic Elderly Subjects","type":"article-journal","volume":"28"},"uris":["http://www.mendeley.com/documents/?uuid=8c40e15e-ae09-473f-a3ef-36228ef41911"]}],"mendeley":{"formattedCitation":"[1]","plainTextFormattedCitation":"[1]","previouslyFormattedCitation":"[1]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[1]. It is generally believed that HP infection is one of the most widely distributed chronic infections that often remains asymptomatic , and may possibly release their antigens that provoke a local and systemic inflammatory reactions ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.4330/wjc.v7.i4.187","abstract":"In this review, we discussed the findings and concepts underlying the potential role of Helicobacter pylori (H. pylori ) infections in the initiation, development or persistence of atherosclerosis and coronary heart disease (CHD). This Gram-negative bacterium was described by Marshall and Warren in 1984. The majority of infected subjects carries and transmits H. pylori with no symptoms; however, in some individuals these bacteria may cause peptic ulcers, and even gastric cancers. The widespread prevalence of H. pylori infections and the fact that frequently they remain asymptomatic may suggest that, similarly to intestinal microflora, H. pylori may deliver antigens that stimulate not only local, but also systemic inflammatory response. Recently, possible association between H. pylori infection and extragastric disorders has been suggested. Knowledge on the etiology of atherosclerosis together with current findings in the area of H. pylori infections constitute the background for the newly proposed hypothesis that those two processes may be related. Many research studies confirm the indirect association between the prevalence of H. pylori and the occurrence of CHD. According to majority of findings the involvement of H. pylori in this process is based on the chronic inflammation which might facilitate the CHDrelated pathologies. It needs to be elucidated, if the infection initiates or just accelerates the formation of atheromatous plaque.","author":[{"dropping-particle":"","family":"Chmiela M, Gajewski A","given":"and Rudnicka K.","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"World Journal of Cardiology","id":"ITEM-1","issue":"4","issued":{"date-parts":[["2015"]]},"page":"187-203","title":"Helicobacter pylori vs coronary heart disease - searching for connections","type":"article","volume":"7"},"uris":["http://www.mendeley.com/documents/?uuid=cf8efd7a-f87f-4376-9112-b919a5cc79c5"]}],"mendeley":{"formattedCitation":"[2]","plainTextFormattedCitation":"[2]","previouslyFormattedCitation":"[2]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[2], thus in the last several decades many studies suggested a possible link between HP and extra gastric diseases including CAD ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.4330/wjc.v7.i4.187","abstract":"In this review, we discussed the findings and concepts underlying the potential role of Helicobacter pylori (H. pylori ) infections in the initiation, development or persistence of atherosclerosis and coronary heart disease (CHD). This Gram-negative bacterium was described by Marshall and Warren in 1984. The majority of infected subjects carries and transmits H. pylori with no symptoms; however, in some individuals these bacteria may cause peptic ulcers, and even gastric cancers. The widespread prevalence of H. pylori infections and the fact that frequently they remain asymptomatic may suggest that, similarly to intestinal microflora, H. pylori may deliver antigens that stimulate not only local, but also systemic inflammatory response. Recently, possible association between H. pylori infection and extragastric disorders has been suggested. Knowledge on the etiology of atherosclerosis together with current findings in the area of H. pylori infections constitute the background for the newly proposed hypothesis that those two processes may be related. Many research studies confirm the indirect association between the prevalence of H. pylori and the occurrence of CHD. According to majority of findings the involvement of H. pylori in this process is based on the chronic inflammation which might facilitate the CHDrelated pathologies. It needs to be elucidated, if the infection initiates or just accelerates the formation of atheromatous plaque.","author":[{"dropping-particle":"","family":"Chmiela M, Gajewski A","given":"and Rudnicka K.","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"World Journal of Cardiology","id":"ITEM-1","issue":"4","issued":{"date-parts":[["2015"]]},"page":"187-203","title":"Helicobacter pylori vs coronary heart disease - searching for connections","type":"article","volume":"7"},"uris":["http://www.mendeley.com/documents/?uuid=cf8efd7a-f87f-4376-9112-b919a5cc79c5"]}],"mendeley":{"formattedCitation":"[2]","plainTextFormattedCitation":"[2]","previouslyFormattedCitation":"[2]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[2]. Although some studies established such association between HP infection and CAD but others did not  ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1016/j.carpath.2011.09.011","ISSN":"10548807","abstract":"Introduction: Several epidemiological studies have proposed an association between Helicobacter pylori infection and coronary artery disease. In the current study, we aimed to evaluate the prevalence and relevance of H. pylori infection, using polymerase chain reaction (PCR) methods, in the coronary arterial wall of Iranian patients who have already undergone coronary bypass grafting (CABG). Methods: A total of 105 consecutive patients who underwent CABG at the Department of Cardiovascular Surgery of Baqiyatallah University of Medical Sciences were included in the study, and biopsy specimens from their coronary plaques were taken and analyzed using the PCR methods for detecting Helicobacter species (H Spp.). Fifty-three specimens from biopsies of the left internal mamillary artery in the same patients were also collected and tested. Results: H. Spp. PCR test result was positive for 31 (29.5%) specimens from coronary artery atherosclerotic plaques. Serologic test results also showed 25 (23.8%) positive cases for H. pylrori immunoglobulin A (IgA) and 56 (53.3%) positive for anti-H. pylori immunoglobulin G. None of the specimens from the mamillary artery were positive for H Spp. genome when it was evaluated using PCR (P<.0001). Patients with positive test result for H. pylori IgA were significantly more likely to have higher total cholesterol and low-density lipoprotein (LDL) levels than IgA-negative patients. Conclusion: H Spp. infection replication in the coronary arterial wall is associated with atherosclerotic plaque formation. Seropositivity for H. pylori IgA may also enhance blood values of total cholesterol and LDL in these patients. � 2012 Elsevier Inc. All rights reserved.","author":[{"dropping-particle":"","family":"Izadi","given":"Morteza","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fazel","given":"Mojgan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Sharubandi","given":"Seyed Hossein","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Saadat","given":"Seyed Hassan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Farahani","given":"Maryam Moshkani","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Nasseri","given":"Mohammad Hassan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Dabiri","given":"Hossein","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Safiaryan","given":"Reza","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Esfahani","given":"Ali Akbar","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ahmadi","given":"Ali","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Jafari","given":"Nematollah Jonaidi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ranjbar","given":"Reza","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Jamali-Moghaddam","given":"Saeed Reza","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kazemi-Saleh","given":"Davood","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kalantar-Motamed","given":"Mohammad Hassan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Taheri","given":"Saeed","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Cardiovascular Pathology","id":"ITEM-1","issue":"4","issued":{"date-parts":[["2012"]]},"page":"307-311","title":"Helicobacter species in the atherosclerotic plaques of patients with coronary artery disease","type":"article","volume":"21"},"uris":["http://www.mendeley.com/documents/?uuid=be0c8a42-e598-4f64-9cf1-91dfa405c0da"]},{"id":"ITEM-2","itemData":{"DOI":"10.2147/TCRM.S142193","ISSN":"1178203X","abstract":"� 2017 Jukic et al. Aim: Controversy exists concerning the relation between Helicobacter pylori (HP) infection and coronary artery disease (CAD). We aimed to examine the relationship between HP infection and severity of coronary atherosclerosis in patients with chronic CAD. Patients and methods: A total of 150 patients (109 [73%] men; mean age 62.61�10.23 years) scheduled for coronary artery bypass grafting surgery were consecutively enrolled in the cross-sectional study. According to rapid urease test and/or gastric biopsy samples stained with hematoxylin and eosin and according to Giemsa, patients were classified as HP positive (n=87; 58%) or HP negative (n=63; 42%). Coronary angiograms were scored by quantitative assessment, using multiple angiographic scoring system: 1) vessel score (number of coronary arteries stenosed e"50%), 2) Gensini score (assigning a severity score to each coronary stenosis according to the degree of luminal narrowing and its topographic importance) and 3) angiographic severity score (number of coronary artery segments stenosed e"50%). Results: In comparison to HP-negative patients, HP-positive patients were more frequently hypertensive (P=0.014), had higher values of systolic (P=0.043) and diastolic (P=0.005) blood pressure and total cholesterol (P=0.013) and had lower values of high-density lipoprotein-cholesterol (HDL-C; P=0.010). There were no significant differences between the groups in the severity of coronary atherosclerosis: vessel score (P=0.152), Gensini score (P=0.870) and angiographic severity score (P=0.734). Conclusion: It is likely that HP infection is not a risk factor for the severity of coronary atherosclerosis in chronic CAD patients.","author":[{"dropping-particle":"","family":"Jukic","given":"Anita","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bozic","given":"Dorotea","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kardum","given":"Dusko","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Becic","given":"Tina","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Luksic","given":"Bruno","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Vrsalovic","given":"Mislav","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ljubkovic","given":"Marko","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fabijanic","given":"Damir","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Therapeutics and Clinical Risk Management","id":"ITEM-2","issued":{"date-parts":[["2017"]]},"page":"933-938","title":"Helicobacter pylori infection and severity of coronary atherosclerosis in patients with chronic coronary artery disease","type":"article","volume":"13"},"uris":["http://www.mendeley.com/documents/?uuid=3bcfe914-38b8-4c29-a971-ed923e3d0480"]}],"mendeley":{"formattedCitation":"[3,4]","plainTextFormattedCitation":"[3,4]","previouslyFormattedCitation":"[3,4]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[3,4].
Macrophage migration inhibitory factor was recognized as pro-inflammatory and pro-atherosclerotic factor that triggers formation of foam cell, development and destabilization of plaque lesions. Previous studies demonstrated that macrophage and many other cell types express MIF constitutively, and  release it rapidly in response to stimulation by gram negative and positive bacterial toxins, and cytokine ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1016/j.atherosclerosis.2014.09.010","ISBN":"1879-1484","ISSN":"18791484","PMID":"25463068","abstract":"Objective: Monocyte infiltration is a critical step in the pathophysiology of plaque instability in coronary artery disease (CAD). Macrophage migration inhibitory factor (MIF) is involved in atherosclerotic plaque progression and instability leading to intracoronary thrombosis. Gremlin-1 (Grem1) has been recently identified as endogenous inhibitor of MIF. To date there are no data on the clinical impact of this interaction in cardiovascular patients. Methods and results: Plasma levels of MIF and Grem1 were determined by enzyme-linked immunoassay in patients with acute coronary syndromes (ACS, n=120; stable CAD, n=166 and healthy control subjects, n=25). MIF levels were significantly increased in ACS compared to stable CAD and healthy control (ACS: median 2.85; IQR 3.52ng/ml; versus SAP: median 1.22; IQR 2.99ng/ml; versus healthy control: median 0.10; IQR 0.09ng/ml, p<0.001). Grem1 levels were significantly higher in ACS and stable CAD patients compared to healthy control (ACS: median 211.00; IQR 130.47ng/ml; SAP: median 220.20; IQR 120.93ng/ml, versus healthy control: median 90.57; IQR 97.68ng/ml, p<0.001). Grem1/MIF ratio was independently associated with ACS, whereas the single parameters were not associated with the presence of ACS. Furthermore, Grem1/MIF ratio was associated with angiographic signs of intracoronary thrombi and severity of thrombus burden. Conclusion: These novel findings suggest a potential role of Grem1/MIF ratio to indicate acuity of CAD and the grade of plaque stability. Prospective angiographic cohort studies involving plaque imaging techniques are warranted to further characterize the prognostic role of this novel risk marker in CAD patients.","author":[{"dropping-particle":"","family":"M�ller","given":"Iris I.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"M�ller","given":"Karin A L","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Karathanos","given":"Athanasios","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Sch�nleber","given":"Heiko","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Rath","given":"Dominik","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Vogel","given":"Sebastian","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Chatterjee","given":"Madhumita","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schmid","given":"Martina","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Haas","given":"Maximilian","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Seizer","given":"Peter","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Langer","given":"Harald","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schaeffeler","given":"Elke","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schwab","given":"Matthias","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Gawaz","given":"Meinrad","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Geisler","given":"Tobias","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Atherosclerosis","id":"ITEM-1","issue":"2","issued":{"date-parts":[["2014"]]},"page":"426-432","title":"Impact of counterbalance between macrophage migration inhibitory factor and its inhibitor Gremlin-1 in patients with coronary artery disease","type":"article-journal","volume":"237"},"uris":["http://www.mendeley.com/documents/?uuid=fa347cd1-3dea-4fd1-a643-ebfdf1a30bb2"]},{"id":"ITEM-2","itemData":{"DOI":"10.1080/00365540310016277","ISSN":"0036-5548","PMID":"14620137","abstract":"Among innate immune cells, macrophages play an essential role in the sensing and elimination of invasive microorganisms. Binding of microbial products to pathogen-recognition receptors stimulates macrophages to release cytokines and other effector molecules that orchestrate the host innate and adaptive immune responses. Recently, the protein known as macrophage migration inhibitory factor (MIF) has emerged as a pivotal mediator of innate immunity. First identified as a T-cell cytokine, MIF was rediscovered as a protein released by pituitary cells after exposure to endotoxin [lipopolysaccharide (LPS)] or bacteria and in response to stress. Monocytes, macrophages and lymphocytes constitutively express MIF, which is rapidly released after stimulation with bacterial endotoxins and exotoxins, and cytokines. MIF induces powerful proinflammatory biological responses and has been shown to be an important effector molecule of septic shock. High levels of MIF have been detected in the circulation of patients with severe sepsis and septic shock. Inhibition of MIF activity with neutralizing anti-MIF antibodies or deletion of the Mif gene led to a marked reduction in cytokine production and protected mice from lethal bacterial sepsis and toxic shock induced by Gram-negative endotoxin or Gram-positive exotoxins. Investigations into the mechanisms whereby MIF modulates innate immune responses to endotoxin and Gram-negative bacteria have shown that MIF up-regulates the expression of Toll-like receptor 4 (TLR4), the signal-transducing molecule of the LPS receptor complex. Thus, MIF enables cells, such as the macrophage, that are at the forefront of the host antimicrobial defences, to sense promptly the presence of invading Gram-negative bacteria and mount an innate immune response. Given that it is a pivotal regulator of innate immune responses to bacterial infections, MIF appears to be a perfect target for novel therapeutic interventions in patients with severe sepsis.","author":[{"dropping-particle":"","family":"Calandra T","given":"","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Scandinavian journal of infectious diseases","id":"ITEM-2","issue":"9","issued":{"date-parts":[["2003"]]},"page":"573-576","title":"Macrophage migration inhibitory factor and host innate immune responses to microbes.","type":"article","volume":"35"},"uris":["http://www.mendeley.com/documents/?uuid=9c8c4565-60ed-4e30-b735-d24b2a33cda2"]}],"mendeley":{"formattedCitation":"[5,6]","plainTextFormattedCitation":"[5,6]","previouslyFormattedCitation":"[5,6]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[5,6].
The association between MIF and HP has been studied in many diseases ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1086/421915","ISSN":"0022-1899","abstract":"The macrophage migratory inhibitory factor (MIF) plays a pivotal role in inflammatory and immune diseases; however, its role in gastrointestinal diseases has not been clarified. This study intended to determine the expression of MIF, by gastric epithelial cells, T cells, and macrophages, in Helicobacter pylori�induced gastritis. Sixty-four patients (30 males, 34 females; mean age, 47 years) referred for upper endoscopy were recruited. Biopsy specimens from the gastric antrum and corpus were obtained for (1) detection of H. pylori and histological examination, (2) single and double immunostaining to test for expression of MIF protein in epithelial cells, T cells, and macrophages, and (2) in situ hybridization for expression of MIF mRNA within the lamina propria. In mucosal specimens from each of the 2 sites, both the percentage of MIF + epithelial cells and the numbers of MIF mRNA + inflammatory cells, MIF + T cells, and MIF + macrophages were significantly higher in H. pylori�positive patients than in H. pylori�negative patients. Overall, the percentage of MIF + epithelial cells and the numbers of MIF mRNA + cells, MIF + T cells, and MIF + macrophages were higher in the antrum than in the corpus. The percentage of MIF + epithelial cells and the numbers of MIF mRNA + cells, MIF + T cells, and MIF + macrophages increased in chronic gastritis, but, in the absence of H. pylori infection, this increase disappeared for all except MIF + T cells. Therefore, H. pylori infection is associated with increased expression of the MIF protein and MIF mRNA in gastric epithelial and inflammatory cells; along with other cytokines, MIF may play a significant role in gastric inflammation related to H. pylori infection.","author":[{"dropping-particle":"","family":"Xia","given":"Harry�Hua Xiang","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lam","given":"Shiu Kum","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Huang","given":"Xiao Ru","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Wong","given":"Wai Man","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Leung","given":"Suet Yi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Yuen","given":"Siu Tsan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lan","given":"Hui Yao","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Wong","given":"Benjamin�Chun Yu","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"The Journal of Infectious Diseases","id":"ITEM-1","issue":"2","issued":{"date-parts":[["2004"]]},"page":"293-302","title":"Helicobacter pylori Infection Is Associated with Increased Expression of Macrophage Migratory Inhibitory Factor�by Epithelial Cells, T Cells, and Macrophages�in Gastric Mucosa ","type":"article","volume":"190"},"uris":["http://www.mendeley.com/documents/?uuid=4a92369f-9fc1-4a89-9f1f-095ed2177422"]},{"id":"ITEM-2","itemData":{"DOI":"10.1590/s1807-59322010000800010","ISSN":"1807-5932","abstract":"OBJECTIVES: To determine the effect of Helicobacter pylori (H. pylori) eradication on blood levels of high-sensitivity C-reactive protein (hs-CRP), macrophage migration inhibitory factor and fetuin-A in patients with dyspepsia who are concurrently infected with H. pylori. METHODS: H.pylori infection was diagnosed based on the 14C urea breath test (UBT) and histology. Lansoprazole 30 mg twice daily, amoxicillin 1 g twice daily, and clarithromycin 500 mg twice daily were given to all infected patients for 14 days; 14C UBT was then re-measured. In 30 subjects, migration inhibitory factor, fetuin-A and hs-CRP levels were examined before and after the eradication of H. pylori infection and compared to levels in 30 healthy subjects who tested negative for H. pylori infection. RESULTS: Age and sex distribution were comparable between patients and controls. Migration inhibitory factor and hs-CRP levels were higher, and fetuin-A levels were lower, in H. pylori-infected patients (p<0.05). Following eradication of H. pylori, migration inhibitory factor and hs-CRP levels were significantly decreased, whereas fetuin-A levels were increased. However, eradication of the organism did not change lipid levels (p>0.05). CONCLUSION: These findings suggest that H. pylori eradication reduces the levels of pro-inflammatory cytokines such as migration inhibitory factor and hs-CRP and also results in a significant increase in anti-inflammatory markers such as fetuin-A.","author":[{"dropping-particle":"","family":"Kebapcilar","given":"Levent","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bilgir","given":"Oktay","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Cetinkaya","given":"Elif","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Akyol","given":"Murat","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bilgir","given":"Ferda","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bozkaya","given":"Giray","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Clinics","id":"ITEM-2","issue":"8","issued":{"date-parts":[["2010"]]},"page":"799-802","title":"The effect of Helicobacter pylori eradication on macrophage migration inhibitory factor, C-reactive protein and fetuin-a levels","type":"article","volume":"65"},"uris":["http://www.mendeley.com/documents/?uuid=d29240ee-c5f0-4175-b9c6-559883b7e66b"]},{"id":"ITEM-3","itemData":{"DOI":"10.3748/wjg.v21.i34.9916","ISSN":"22192840","abstract":"� The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved. � 2015 Baishideng Publishing Group Inc. All rights reserved. AIM: To investigate macrophage migration inhibitory factor (MIF) expression and its clinical relevance in gastric cancer, and effects of MIF knockdown on proliferation of gastric cancer cells. METHODS: Tissue microar ray containing 117 samples of gastric cancer and adjacent non-cancer normal tissues was studied for MIF expression by immunohistochemistry (IHC) semiquantitatively, and the association of MIF expression with clinical parameters was analyzed. MIF expression in gastric cancer cell lines was detected by reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blot. Two pairs of siRNA targeting the MIF gene (MIF si-1 and MIF si-2) and one pair of scrambled siRNA as a negative control (NC) were designed and chemically synthesized. All siRNAs were transiently transfected in AGS cells with OligofectamineTM to knock down the MIF expression, with the NC group and mock group (Oligofectamine� alone) as controls. At 24, 48, and 72 h after transfection, MIF mRNA was analyzed by RTPCR, and MIF and proliferating cell nuclear antigen (PCNA) proteins were detected by Western blot. The proliferative rate of AGS cells was assessed by methylthiazolyl tetrazolium (MTT) assay and colony forming assay. RESULTS: The tissue microarray was informative for IHC staining, in which the MIF expression in gastric cancer tissues was higher than that in adjacent noncancer normal tissues (P  <  0.001), and high level of MIF was related to poor tumor differentiation, advanced T stage, advanced tumor stage, lymph node metastasis, and poor patient survival (P  <  0.05 for all). After siRNA transfection, MIF mRNA was measured by real-time PCR, and MIF protein and PCNA were assessed by Western blot analysis. We found that compared to the NC group and mock group, MIF expression was knocked down successfully in gastric cancer cells, and PCNA expression was downregulated with MIF knockdown as well. The cell counts and the doubling times were assayed by MTT 4 d after transfection, and colonies formed were assayed by colony forming assay 10 d after transfection; all these showed significant changes in gastric cancer cells transfected with specific siRNA compared with the control siRNA and mock groups (P  <  0.001 for all). CONCLUSION: MIF could be of prognostic value in gastric cancer and might be a potential target for small-�","author":[{"dropping-particle":"","family":"He","given":"Long Jun","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Xie","given":"Dan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Hu","given":"Pin Jin","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Liao","given":"Yi Ji","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Deng","given":"Hai Xia","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kung","given":"Hsiang Fu","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Zhu","given":"Sen Lin","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"World Journal of Gastroenterology","id":"ITEM-3","issue":"34","issued":{"date-parts":[["2015"]]},"page":"9916-9926","title":"Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer","type":"article","volume":"21"},"uris":["http://www.mendeley.com/documents/?uuid=84d98f66-d27f-41cb-aaf9-9f9d4dd8612a"]}],"mendeley":{"formattedCitation":"[7�9]","plainTextFormattedCitation":"[7�9]","previouslyFormattedCitation":"[7�9]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[7�9]. Therefore, this study aimed to find if there is association between HP seropositivity and MIF level in MI patients admitted for PPCI which may reflect a possible impact of HP infection on MI severity. 
 


Materials and methods
This case control study included 176 subjects who were admitted to surgical specialty hospital-cardiac centre in Erbil City, Iraq from January to December 2017. Based on the clinical, laboratory and coronary angiography, participants were classified to 100 MI according to WHO criteria ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1093/ije/dyq165","ISSN":"03005771","abstract":"WHO has played a leading role in the formulation and promulgation of standard criteria for the diagnosis of coronary heart disease and myocardial infarction since early 1970s.","author":[{"dropping-particle":"","family":"Mendis","given":"Shanthi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Thygesen","given":"Kristian","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kuulasmaa","given":"Kari","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Giampaoli","given":"Simona","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Mahonen","given":"Markku","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Blackett","given":"Kathleen Ngu","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lisheng","given":"Liu","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"International Journal of Epidemiology","id":"ITEM-1","issue":"1","issued":{"date-parts":[["2011"]]},"page":"139-146","title":"World Health Organization definition of myocardial infarction: 2008-09 revision","type":"article","volume":"40"},"uris":["http://www.mendeley.com/documents/?uuid=5ad9b23d-0951-42ed-a028-14363780085c"]}],"mendeley":{"formattedCitation":"[10]","plainTextFormattedCitation":"[10]","previouslyFormattedCitation":"[10]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[10], 38 SAP, and 38 subjects confirmed to have normal coronary angiography (COA) with normal range of creatine kinase-MB (CKMB), and high sensitivity cardiac troponin T(hscTnT). They were admitted for chest pain, later found not linked to cardiac origin.
Diagnosis of SAP was based on significant stenosis (>30) documented by COA, with no present change in the character, incidence, or chest pain severity. None of the MI or SAP patients were referred for coronary bypass graft (CABG). 
The study excluded any case with active inflammation, autoimmune, recent trauma or surgery. Cases with heart failure, cardiomyopathy, congenital or sever valvular heart diseases, stroke, post MI were also excluded from the study.
 Routine biochemical and haematological tests were done for all the participants as baseline laboratory investigations. Demographic information, smoking status, diabetes, hypertension, and history of using medications were obtained by direct interviewing with the participants using a well-designed questionnaire. Diagnosis of hypertension (HTN) was based on either regularly repeated measurements of blood pressure e"140/90	 mmHg (at least two to three times in stable situation) or on previous diagnosis or taking anti hypertension therapy at present.  Diabetes mellitus (DM) was diagnosed according to haemoglobin A1c (HbA1c) level >6.5% ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1016/j.diabres.2011.03.012","ISSN":"01688227","abstract":"The report on the acceptability of glycated haemoglobin, or HbA1c, as an additional test to diagnose this debilitating and deadly disease.","container-title":"Diabetes Research and Clinical Practice","id":"ITEM-1","issue":"3","issued":{"date-parts":[["2011"]]},"page":"299-309","title":"Use of glycated haemoglobin (HbA1c) in the diagnosis of diabetes mellitus","type":"article","volume":"93"},"uris":["http://www.mendeley.com/documents/?uuid=eb5c4b73-6cb7-4a08-983c-89d333211466"]}],"mendeley":{"formattedCitation":"[11]","plainTextFormattedCitation":"[11]","previouslyFormattedCitation":"[11]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[11], or on the previous history of diabetes who either treated by medications or controlled by diet. Calculation of body mass index (BMI) was based on the standard equation weight (Kg) /height m2. 
COA was done for all participants through right femoral artery. Patient underwent PCI if result of angiography revealed new arterial occlusion. Numbers of diseased vessels were also identified depending on COA results.
Blood samples were collected immediately after admission and before COA in different Vacutainer tubes. Serum and plasma were separated by centrifugation and stored at -80�%C for further analysis. Additionally, 3 ml of whole blood were obtained directly from femoral artery during COA for molecular study. 
Using quantitative sandwich ELISA kits to measure plasma levels of MIF (Mybiosource, USA), anti HP-IgM (IBL- international GMBH, Germany), IgG (Accu-Bind, USA) and IgA (Accu-Bind, USA). Concentration of plasma fibrinogen was measured by Clauss method using FIBRI-PREST AUTOMATE (Diagnostica Stago, France). Levels of serum high sensitivity C-reactive protein (hsCRP) were determined using particle enhanced immuno turbidimetric assay by cobas c111 (Roche Diagnostics GmbH). Other investigation (biochemical and haematological) were done according to the manufacture instructions. Based on the kit s reference cut off values, levels of both anti HP-IgG and IgA>20U/ml were considered positive. For IgM, levels <8U/ml, and >12U/ml unit were considered as negative and positive respectively, values between 8U/ml and 12U/ml were considered as equivocal.
Positive results of anti HP-IgM(IgM+) and/or IgA+IgG+ with high titres were further analysed by PCR for HP genomic material 16SrRNA. A total of 28 participants had equivocal levels of anti-HP�IgM, and they were designated to have negative results of HP.
DNA was extracted according to manufacturer�s instructions kit (Promega, USA). The quality and quantity of extracted DNA was estimated using spectrophotometry (Labtech,UK), and 1% agarose gel electrophoresis.  For PCR amplification of 500 bp fragment of bacterial genome16SrRNA a specifically designed primers were used: Forward primer 5-GCTAAGAGATCAGCCTATGTCC-3, and reverse primers 5-TGGCAATCAGCGTCAGGTAATG-3ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"doi:10.1155/2012/245167","PMID":"22778721","abstract":"Helicobacter pylori is recognized by the World Health Organization to be the primary cause of peptic ulcers, chronic gastritis, and stomach cancer, though the source of human infection is not well understood. One of the problems in understanding the source of human contamination is the difficulty in isolating the organism from the environment. However, the combination of PCR results with those of culturing of 471 drinking water samples can provide a more accurate picture of H. pylori detection. In this method 78 presumptive H. pylori colonies out of 266 tap water samples were obtained in the preliminary detection on modified Columbia agar (MCUA) slant relying on urease positivity with a rate of 29.3%. However, only 11 out of them were confirmed by Gram staining and biochemical tests reducing the rate to 4.13% whereas only 3 (1.46%) from 205 reverse osmosis (RO) water samples. Furthermore, only 6 (54.5%) out of the 11 isolates from tap water and 1 (33.3%) of the 3 RO isolates were confirmed by 16SrRNA PCR. Thus PCR confirmation reduced the rate to 2.2%. In addition, only 4 (4%) of 100 tap water samples negative for H. pylori by culture method were H. pylori positive by 16SrRNA. Water samples were collected from 24 districts of Basrah Governorate from February�December 2009. The direct recovery of H. pylori from drinking water is both alarming and scientifically exciting in terms of the investigation of its epidemiology.","author":[{"dropping-particle":"","family":"Al-Sulami AA, Al-Edani TAA","given":"Al-Abdula AA","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Gastroenterology Research and Practice","id":"ITEM-1","issued":{"date-parts":[["2012"]]},"page":"5 pages","title":"Culture Method and PCR for the Detection of Helicobacter pylori in Drinking Water in Basrah Governorate Iraq","type":"article-journal"},"uris":["http://www.mendeley.com/documents/?uuid=6cae99c0-115c-4dde-9223-14841b263a29"]}],"mendeley":{"formattedCitation":"[12]","plainTextFormattedCitation":"[12]","previouslyFormattedCitation":"[12]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[12]. In each run of PCR, human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control to validate the quality of extraction by using a set of specific primers which provided by BIOLAB Company/Iraq: Forward primer 5-CCAGAACATCATCCCTGCCT-3, and reverse primer 5-AAGATGAAAAGAGTTGTCAGGGC-3, to amplify 430 bp fragment of GAPDH. Positive and negative controls were also included in each reaction. For positive control, an HP DNA was used, and provided by WahjDNA Company/Iraq. Negative control contained all PCR reagents except DNA tested sample. For positive control amplification, GAPDH primers were excluded while for GAPDH amplification, HP primers were excluded. For each sample, PCR amplification was carried out in a 20�L of reaction mixture containing 10�L of master mix, 1�L of forward primer, 1�L of reverse primer for both HP 16SrRNA and GAPDH, 1�L DNA samples, and 5�L Nuclease free water. Amplification reaction was achieved in an automated thermocycler (TECHNE, UK) including: Initial denaturation step at 95�%C for 5 minutes (min), followed by 35 cycles denaturation at 95�%C for 45 seconds (sec), annealing at 57�%C for 30 sec and extension at 72�%C for 45 sec, and an additional extension step at 72�%C for 5 min. PCR products were electrophoresed in 1% agarose.
The study protocol was approved by medical ethic committee in the College of Medicine/ Hawler Medical University.
Statistical analysis
Data analysis was carried out by using SPSS 23.0 software. One way analysis of variance (ANOVA) was used for Continuous data represented as mean and standard error, while chi-squared test or fisher exact test were used for categorical data. Value of P<0.05 was considered statistically significant.   
Results 
The demographic characteristics and clinical findings of the participants are illustrated in Table 1. Anti Hp-IgG was positive in 68%, 64.49%, and 55.26% of MI, all CAD, and SAP patients respectively. Control group showed higher percentage (84.21%) than the patient groups. In general male gender, smokers, and hypertensive individuals were found predominantly in group seropositive patients with statistically significant differences compared to control group (p<0.05). Marriage status revealed significant difference between seropositive and negative subjects within same group (p<0.05).  Patients with group age d"45 included less seropositive while the higher proportion of seropositive patients were found in age group 46-55 years and 56-65 years with significant difference (p<0.05). A statistically significant difference was observed between seropositive and seronegative MI patients regarding gender, smoking, HTN, marriage status and number of diseased vessels (p<0.05). 








Table 1: Demographic and clinical features of patient and control groups based on anti HP-IgG positive and negative results.
All CAD 
(MI+SAP)
138MI
100SAP
38Control
38P valueNo (%)Seronegative
49(35.51)Seropositive
89(64.49)Seronegative
32(32)Seropositive
68(68)Seronegative
17(44.74)Seropositive
21(55.26)Seronegative
6(15.79)Seropositive
32(84.21)Male 
Female 25(51.02)
24(48.98)71(79.55)7.
18(20.45)20(62.5)
12(37.5)56(82.35)7.
12(17.65)5(29.41)
12(70.59)15(71.43)7.
6(28.57)
4(66.67
2(33.33)
18(56.25)
14(43.75)
*0.002
**0.023
***0.067Smoker
Non-smoker26(53.06)
23(46.94)69(77.53)7.
20(22.47)18(56.25)
14(43.75)54(79.41)7.
14(20.59)8(47.06)
9(52.94)15(71.43)
6(28.57)2(33.33)
4(66.67)14(43.75)
18(56.25)*0.001
**0.001
***0.160HTN
Non-HTN27(55.10)
22(44.9)  54(60.67)
35(39.33)12(37.5)
20(62.5)  40(58.82)7.
28(41.18)   15(88.24)
2(11.76)14(61.90)
7(38.1)0(0)
6(100)12(37.5)
20(62.5)*0.005
**0.007
***0.001DM
Non-DM24(48.98)
25(51.02)36(40.45) 
53(59.55)18(56.25)
14(43.75)28(41.18)
40(58.82)6(35.29)
11(64.71)8(38.1)
13(61.90)--0.5748.

Marriage status
Single
Married
Widow+ divorce
 3(6.12) 7.
39(79.59)
7(14.29)
2(2.25)
75(84.27)
12(13.48)
1(3.12)
27(84.38)
4(12.5)
2(2.94) 7.
54(79.41)
12(17.65)
2(11.76)
12(70.59)
3(17.65)
0(0)7.
21(100)
0(0)
0(0)
6(100)
0(0)
1(3.13) 7.
25(78.12)
6(18.75)
*0.796
**0.962
***0.097Education Levels
Illiterate 
Primary + secondary 
High School and above
   21(42.86) 
16(32.65)
12(24.49)
43(48.31)
32(35.96)
14(15.73)
12(37.5)
14(43.75)
6(18.75)
30(44.11)
26(38.24)
12(17.65)
9(52.94)
2(11.76)
6(35.3)
13(61.91)
6(28.57)
2(9.52)
4(66.67)
0(0)
2(33.33)
12(37.5)
16(50)
4(12.5)
*0.207
**0.357
***0.022Employment Situation
Employed
Un-employed
Retired
      8(16.33)
39(79.59)
2(4.08)
  18(20.22)
58(65.17)
13(14.61)
   4(12.5)
26(81.25)
2(6.25)
  12(17.65)
45(66.17)
11(16.18)
   4(23.53)
13(76.47)
0(0)
     6(28.57)
13(61.91)
2(9.52)
    2(33.33)
3(50)
1(16.67)
  4(12.5)
27(84.37)
1(3.13)
*0.122
**0.201
***0.219Age groups (years)
d"45
46-55
56-65
e"66
8(16.33)
20(40.82)
11(22.45)
10(20.40)
6(6.74)
33(37.08)
26(29.21)
24(26.97)
8(25)
10(31.25)
8(25)
6(18.75)
6(8.82)
20(29.41)
20(29.41)
22(32.35)
0(0)
10(58.82)
3(17.65)
4(23.53)
0 (0)
13(61.91)
6(28.57)
2(9.52)
2(33.33)
2(33.33)
2(33.33)
0(0)
8(25)
10(31.25)
14(43.75)
0(0)
*0.002
**0.002
***0.001No. of diseased vessels
1 vessel
2 vessels
3 vessels
14(28.57)
16(32.65)
19(38.78)
17(19.10)7.
9(10.11)
63(70.79)
4(12.5)
14(43.75)
14(43.75)
11(16.18)7.
6(8.82)
51(75)
10(58.82)
2(11.77)
5(29.41)
6(28.57)
3(14.29)
12(57.14)--

0.0018.Medication 
Aspirin 
Ticagrelor 
Clopidogrel
Anti-diabetes 
Beta blocker 
Statin 
45(91.83)
35(71.43)
16(32.65)
22(44.89)
37(75.51)
41(83.67)
85(95.51)
54(60.67)
35(39.33)
34(38.20)
61(68.54)
67(75.28)
28(87.5)
24(75)
7(21.88)
16(50)
26(81.25)
28(87.5)
64(94.12)
48(70.59)
20(29.41)
26(38.24)
49(72.06)
54(79.41)
17(100)
11(64.71)
9(52.94)
6(35.29)
11(64.71)
13(76.47)
21(100)
6(28.57)
15(71.43)
8(38.1)
12(57.14)
13(61.90)
-
-
-
-
0(0)
-
-
-
-
-
8(25)
-HP; Helicobacter pylori, IgG; immunoglobulin G, CAD; coronary artery disease, MI; myocardial infarction, SAP; stable angina pectoris, DM; diabetes mellitus, HTN; hypertension
Tests of significance between groups (chi-square test, and fisher exact test).
Significance of P value was as following: *all CAD vs control, **MI vs control, ***SAP vs control, 
7. Significant P value (p<0.05) between seropositive and seronegative within the same group. 
8. P value for all CAD vs MI vs SAP.
Table 2 represents the prevalence of anti HP-IgA and IgM in patients and control groups. Group MI showed higher proportion of anti HP-IgA+ followed by all CAD, and SAP (42%, 38.41%, 28.95%) respectively, while control group showed 31.58% IgA seropositivity. Similar results were observed regarding anti HP-IgM+ MI 7%, all CAD 6.52%, SAP 5.26% and control 5.26%). The higher percentage of IgG+IgA+ was found in MI group (50%) followed by all CAD, SAP, and control groups (47.19%, 38.1% and 31.25%) respectively. Likewise, higher percentage of IgG+IgM+ was found in MI (10.29%) followed by all CAD, SAP, and controls (10.11, 9.52%, and 6.25 respectively). 
Table 2: Distribution of anti HP-IgA and anti HP-IgM based on anti HP-IgG positive and negative results.
All-CAD (MI+SAP)
Total
138 MI
Total
100SAP
Total
38Control 
Total
38IgG+
89(64.49)IgG-
49(35.51)IgG+
68(68)IgG-
32(32)IgG+
21(55.26)IgG-
17(44.74)IgG+
32(84.21)IgG-
6(15.79)IgA+
IgA-42(47.19)
47(52.81)11(22.45)
38(77.55)53(38.41)
85(61.59)34(50)
34(50)8(25)
24(75)42(42)
58(58)8(38.1)
13(61.90)3(17.65)
14(82.35)11(28.95)
27(71.05)10(31.25)
22(68.75)2(33.33)
4 (66.67)12(31.58)
26(68.42)IgM+
IgM(equivocal)
IgM-9(10.11)
12(13.48)
68(76.41)0(0)
6(12.24)
43(87.76)9(6.52)
18(13.04)
111(80.44)7(10.29)
12(17.65)
49(72.06)0(0)
4(12.5)
28(87.5)7(7)
16(16)
77(77)2(9.52)
0(0)
19(90.48)0(0)
2(11.76)
15(88.24)2(5.26)
2(5.26)
34(89.47)2(6.25)
10(31.25)
20(62.5)0(0)
0(0)
6(100)2(5.26)
10(26.32)
26(68.42)IgM; immunoglobulin M, IgA; immunoglobulin A, IgG; immunoglobulin G.
Seropositive MI group had higher age, TG, LDL-c, LDL/HDL, MIF, fibrinogen, hsCRP, leukocyte and neutrophil counts compared to other studied groups. A significant difference was observed between seropositive MI patients and seropositive and negative control group regarding MIF (p<0.05). Similar results were remarked concerning age, HDL-c, lipid ratios, and other inflammatory biomarkers (fibrinogen, hsCRP, HbA1c, leukocyte count, neutrophil count, and mean platelet volume (MPV) (p<0.05), more details are illustrated in table 3. 


Table 3: Comparison of different variables between patient and control groups based on anti HP-IgG positive and negative results.
Variables All CAD
(MI+SAP)
138MI
100SAP
38Control 
38pMean�SESeronegative
49(35.51)Seropositive
89(64.49)Seronegative
32(32%)Seropositive
68(68%)Seronegative
17(44.74)Seropositive
21(55.26)Seronegative
6(15.79%)Seropositive
32(84.21%)Age (years)55.76�1.76ab59.88�1.67a55.18�2.54a 61.20�1.99b56.77�2.03ab55.40�2.56ab52.0�7.00ab52.5�2.10a0.067BMI(kg/m�)29.16�0.87a28.52�0.71ab30.26�1.03a28.39�0.82abc27.20�1.44abc28.93�1.47abc24.38�1.03bd26.14�0.47cd0.138Lipid profileTC(mg/dl)165.89�8.10 a192.60�13.66a170.63�8.97a191.11�7.26a157.46�16.26a197.66�57.14a122.01�14.93a172.00�12.37a0.478TG(mg/dl)147.91�14.01a161.72�18.51a138.80�15.71a171.62�23.47a164.11�27.62a128.08�13.12a82.55�30.93a128.39�11.08a0.673HDL-c (mg/dl)35.92�1.53a37.97�1.52a34.04�1.76a38.72�1.84a39.26�2.60a35.44�2.34a43.24�7.63 ab50.92�2.56b0.000LDL-c (mg/dl)112.96�7.04ac117.13�5.08a115.36�7.45ac127.35�5.07a108.70�14.99acd82.39�6.92bd63.49�12.35b98.59�8.34bc0.001Non-HDL (mg/dl)129.96�7.63a154.62�13.71a136.59�7.98a152.39�6.94a118.19�15.68a162.21�57.83a78.77�22.36a121.08�13.06a0.359Lipid RatiosTC/ HDL4.72�0.23 ab5.40�0.50 a5.10�0.24 ab5.22�0.29a4.05�0.39 ab6.03�2.03a3.14�0.95ab3.54�0.31b0.163LDL/ HDL3.20�0.19 ac3.25�0.18ac3.47�0.24ac3.49�0.21a2.71�0.29bc2.42�0.27b1.67�0.60b2.03�0.20b0.000Inflammatory MarkersMIF(ng/ml)36.25�4.54ab41.57�3.30a37.60�5.49ab43.49�3.76a33.84�8.38abc35.08�6.86abc17.07�7.57bd21.67�3.89cd0.031Fibrinogen
(mg/dl)416.04�23.43ab466.29�23.91a420.54�27.97ab468.17�28.40a408.05�44.22ab459.90�44.32a337.53�15.06ab349.91�29.43b0.101HsCRP (mg/L)7.25�1.27 ab9.91�2.04a8.91�1.77a11.18�2.52a4.31�1.18ab5.59�2.41ab1.83�0.76ab2.01�0.35b0.112HbA1c%7.27�0.43a7.08�0.32a7.72�0.62a7.12�0.37 a6.46�0.38 ab6.93�0.65ab5.62�0.05ab5.54�0.07b0.085Anti HP-IgM (U/ml)5.31�0.46 a7.66�1.22 a5.19�0.51a8.33�1.53a5.51�0.96a5.38�1.20a6.21�1.30a7.66�1.09a0.484AntiHP-IgA (U/ml)20.75�4.07a30.68�3.67a21.21�5.10a31.38�4.09a19.92�7.15a28.30�8.63a26.08�11.08a32.66�3.67a0.497Heamatological InvestigationsLeukocyte (103/�L)10.33�0.70c11.36�0.54 ac12.26�0.70 ac12.53�0.54 a6.91�0.43 b7.38�0.48 b5.18�0.76b7.00�0.35b0.001Neutrophil (103/�L)8.24�0.65 a9.15�0.59 a9.27�0.67 a9.82�0.59 a4.95�0.40 b4.75�0.38 b3.05�0.1 b4.34�0.63 b0.001Platelet (103/�L)235.52�11.21 ac237.09�7.08 a250.37�15.48 a235.23�7.62 ac209.11�10.63 abc243.40�18.06 ac165.00�7.09 b209.81�13.55 bc0.064MPV (fl)9.01�0.35 ac9.36�0.22 a9.62�0.45a9.39�0.27a7.92�0.29 bc9.28�0.42 ac7.10�0.46 b8.06�0.27 bc0.003Values are presented as mean � SE using ANOVA test, different letters mean significant differences between the groups (p<0.05), in contrary similar letters mean insignificant differences between the groups (p>0.05).
CAD; coronary artery disease, MI; myocardial infarction, SAP; stable angina pectoris, BMI; body mass index, TC; total cholesterol, TG; triglyceride, DHL-c; high density lipoprotein-cholesterol, LDL-c; LDL, low density lipoprotein-cholesterol, MIF; macrophage migration inhibitory factor, hsCRP; high sensitivity C reactive protein, HbA1c; Haemoglobin A1c, Anti HP-IgM;anti helicobacter pylori-immunoglobulin M, Anti HP-IgA; anti helicobacter pylori-immunoglobulin, MPV; mean platelet volume, fL; femtolitre






Table 4 revealed insignificant positive correlation between anti Hp-IgG and MIF (r=0.105, p=0.467), hsCRP(r=0.194, p=0.176), fibrinogen (r=0.004, p=0.980), anti HP-IgA (r=0.207, p=0.148), leukocyte (r=0.103, p=0.477), neutrophil (r=0.135, p=0.349), and MPV (r=0.023, p=0.872).
        Table 4.  Correlation between anti-HP IgG and inflammatory biomarkers in  MI patients
ParametersAnti HP-IgG            rP valueMIF0.1050.467hsCRP0.1940.176Fibrinogen0.0040.980Anti HP-IgA0.2070.148WBC0.1030.477Neutrophil 0.1350.349MPV0.0230.872





















Results of PCR showed no evidence for HP genome, figure 1.


                                Figure 1: PCR assay revealed negative result for HP 16SrRNA genome.
                                M; DNA marker, IC; internal control (GAPDH), PC; positive control, S1, 2, 3, 4; patients 
                                sample,  NC; negative control.                        
                          
                           












Discussion 
Results of current study showed high proportion of positive anti HP-IgG in patients and controls; however, the proportion was remarkably higher in control group. Previous studies also showed a high prevalence of HP infection in asymptomatic Turkish and Iranian healthy individuals 70% and 69% respectivelyADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1016/j.diagmicrobio.2004.07.005","ISSN":"07328893","abstract":"Approximately half of the world population is infected with Helicobacter pylori, particularly in developing countries. The aims of the study were to detect H. pylori infection in asymptomatic Turkish subjects, correlate the infection with the associated risk factors, and to evaluate the cytotoxin-associated gene (CagA) status and other H. pylori antigens. Three hundred nine asymptomatic subjects (124 female) 1-82 years of age (average: 31 years) were serologically tested by enzyme immunoassay and immunoblotting. The enzyme immunoassay detected IgG anti-H. pylori antibodies in sera of 216 (70%) out of 309 subjects, 132 (61%) male. Infection rates of 42% in subjects <10 years of age, 55% in 10-19 years, 66% in 20-29 years, 78% in 30-39 years, 79% in 40-49 years, 91% in 50-59 years, 100% in 60-69 years, and 80% in those >70 years of age were detected. Subjects >45 years of age had significantly higher antibody responses, odds ratio = 0.16 (95% confidence interval: 0.07-0.37), than those <45 years. H. pylori infection was significantly higher in married subjects, odds ratio = 0.38 (95% confidence interval: 0.20-0.73), and those with low socioeconomic status. No correlation between gender, education, smoking, and nonsteroidal anti-inflammatory drug intake and infection was detected. Immunoblots revealed antibodies to CagA in 58 (83%) of 70 samples tested. H. pylori infection is prevalent in the asymptomatic Turkish subjects. Marital and socioeconomic status was significantly associated with the acquisition of H. pylori. Antibodies to CagA antigen were highly prevalent in these subjects. � 2004 Elsevier Inc. All rights reserved.","author":[{"dropping-particle":"","family":"Abasiyanik","given":"M. Fatih","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Tunc","given":"Meral","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Salih","given":"Barik A.","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Diagnostic Microbiology and Infectious Disease","id":"ITEM-1","issue":"3","issued":{"date-parts":[["2004"]]},"page":"173-177","title":"Enzyme immunoassay and immunoblotting analysis of Helicobacter pylori infection in Turkish asymptomatic subjects","type":"article","volume":"50"},"uris":["http://www.mendeley.com/documents/?uuid=e975906b-acb2-49b0-b417-cf1bfe986db8"]},{"id":"ITEM-2","itemData":{"DOI":"10.1111/j.1523-5378.2009.00657.x","ISSN":"10834389","abstract":"Background: Management of Helicobacter pylori, a causative agent of gastrointestinal diseases is an important health pr oblem in most countries. The main reasons include poorly defined epidemiological status and unrecognized mode of bacterial transmission. Our objective was to investigate the prevalence of H. pylori infection in a representative population of Iran and to evaluate possible risk factors for the H. pylori infection. Materials and methods: In this cross-sectional study, 2561 healthy individuals aged 18-65 years (mean age, 35.5 years) were selected out of 12,100,000 inhabitants of Tehran province by cluster sampling. Infection with H. pylori was evaluated by detection of anti-H. pylori IgG antibody in serum. Sociodemographic status of each subject was determined by filling up a questionnaire. Results: Prevalence of H. pylori infection was 69% and was correlated with increasing age. The highest infection rate (79.2%) was seen in individuals 46-55 years old. No association was detected between H. pylori positivity and gender. Low education of the study subjects; low father's and mother's education; poor tooth brushing habit; crowded families in childhood; and lack of household bath, hygienic drinking water, and swage disposal facility in childhood were determined as possible risk factors. Conclusions: The rate of prevalence of H. pylori infection was higher than developed countries. Low socioeconomic status, poor sanitary indications, and crowded families in childhood were related to high prevalence of H. pylori infection in Iran. Accordingly, fecal-oral and oral-oral routes could be considered as the main pathways of transmission of H. pylori. � 2009 The Authors.","author":[{"dropping-particle":"","family":"Nouraie","given":"Mehdi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Latifi-Navid","given":"Saeid","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Rezvan","given":"Houri","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Radmard","given":"Amir Reza","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Maghsudlu","given":"Mahtab","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Zaer-Rezaii","given":"Hanieh","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Amini","given":"Sadigheh","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Siavoshi","given":"Farideh","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Malekzadeh","given":"Reza","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Helicobacter","id":"ITEM-2","issue":"1","issued":{"date-parts":[["2009"]]},"page":"40-46","title":"Childhood hygienic practice and family education status determine the prevalence of Helicobacter pylori infection in Iran","type":"article","volume":"14"},"uris":["http://www.mendeley.com/documents/?uuid=4c54e473-ddbf-4d25-afd0-b43178cd2b87"]}],"mendeley":{"formattedCitation":"[13,14]","plainTextFormattedCitation":"[13,14]","previouslyFormattedCitation":"[13,14]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[13,14].
Infection with HP displays a clear disparity in its prevalence between developed and developing countries. In developed countries, HP prevalence fluctuates between 30% and 50%, while in developing countries the prevalence is much higher and oscillates between 80% and 90% ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1136/gut.34.12.1672","PMID":"8282253","abstract":"The cross sectional study describes the prevalence of infection with Helicobacter pylori as determined by a serodiagnostic assay in over 3000 asymptomatic subjects, in two age groups 25-34 years and 55-64 years, from 17 geographically defined populations in Europe, North Africa, North America, and Japan, using a common protocol for blood collection and serological testing. In all populations combined, the prevalence of infection was higher in the older age group (62�4%) than in the younger age group (34�9%). There was no difference in prevalence of infection between men and women. Subjects with higher education had considerably lower levels of infection (34�1%) compared with subjects with education up to secondary level (46�9%) or those with primary education only (61�6%). This trend was confined to the older of the two age groups. In contrast a trend of increasing prevalence of infection with increasing body mass index was confined to the younger of the two age groups. There was no effect of smoking or alcohol consumption on the prevalence of infection after adjusting for the other risk factors. There was considerable variation in the prevalence of infection between the 17 populations but, within populations, low education standard was consistently and positively associated with the prevalence of infection.","container-title":"Gut","id":"ITEM-1","issue":"12","issued":{"date-parts":[["1993"]]},"page":"1672-1676","title":"Epidemiology of, and risk factors for, Helicobacter pylori infection among 3194 asymptomatic subjects in 17 populations.The EUROGAST Study Group","type":"article-journal","volume":"34"},"uris":["http://www.mendeley.com/documents/?uuid=555ba7d8-8a47-4b07-9d67-110066d24573"]}],"mendeley":{"formattedCitation":"[15]","plainTextFormattedCitation":"[15]","previouslyFormattedCitation":"[15]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[15]. This variation of occurrence  between countries seems to be attributed to the socioeconomic status, and HP may be a measure of health and socioeconomic conditions of the regionADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1111/apt.14561","abstract":"Summary Background: The epidemiology of Helicobacter pylori infection is poorly understood. Aim: To establish the reported regional and national prevalence of H. pylori infection, stratified by age and gender. Methods: All relevant English publications from 2000 to 2017 cited by PubMed and Scopus were retrieved using comprehensive combinations of keywords. The overall prevalence of H. pylori was estimated using both random effect and fixed effect meta-analyses, and presented as prevalence rate (% and 95% CI). The analyses were extended by separation into gender and age groups. Results: A total of 14 056 records were obtained initially. After applying exclusion criteria in several steps, 183 studies were selected. Analysis of 410 879 participants from 73 countries in six continents revealed an overall prevalence of 44.3% (95% CI: 40.9-47.7) worldwide. This rate ranged from 50.8% (95% CI: 46.8-54.7) in developing countries compared with 34.7% (95% CI: 30.2-39.3) in developed countries. The global H. pylori infection rate was 42.7% (95% CI: 39-46.5) in females compared to 46.3% (95% CI: 42.1-50.5) in males. The prevalence in adults (e"18 years) was significantly higher than in children (48.6% [95% CI: 43.8-53.5] vs 32.6% [95% CI: 28.4-36.8], respectively). There was a statistically nonsignificant decrease in the prevalence in 2009-2016 compared with the 2000-2009 period. Conclusions: The observed differences between countries appear to be due to economic and social conditions. H. pylori infection can be a benchmark for the socioeconomic and health status of a country. Further studies are suggested to investigate the natural history of the acquisition of H. pylori infection from childhood into adult life.","author":[{"dropping-particle":"","family":"Zamani","given":"M","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Alizadeh-","given":"W H Miller R","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Alimentary Pharmacology & Therapeutics","id":"ITEM-1","issue":"November 2017","issued":{"date-parts":[["2018"]]},"page":"868-876","title":"Systematic review with meta-analysis : the worldwide prevalence of Helicobacter pylori infection","type":"article-journal","volume":"47"},"uris":["http://www.mendeley.com/documents/?uuid=ced66990-69c4-4938-8172-2589c44ca00f"]}],"mendeley":{"formattedCitation":"[16]","plainTextFormattedCitation":"[16]","previouslyFormattedCitation":"[16]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[16]
Current study investigated the association between anti HP-IgG and MIF in MI patient group compared to SAP and control groups. Overall, MIF level differed insignificantly between seropositive and seronegative subjects within the same group; however, seropositive MI patients demonstrated higher MIF level compared to all study groups.  Meanwhile, seropositive MI patients showed significant elevation of MIF level compared to seropositive and negative control groups. 
Previous studies reported that HP infection stimulate the expression of both MIF mRNA and MIF protein in epithelial and inflammatory cells of gastric mucosa. Consequently, MIF possibly play an important role alongside with various cytokines in gastric inflammation associated with HP infectionADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1086/421915","ISSN":"0022-1899","abstract":"The macrophage migratory inhibitory factor (MIF) plays a pivotal role in inflammatory and immune diseases; however, its role in gastrointestinal diseases has not been clarified. This study intended to determine the expression of MIF, by gastric epithelial cells, T cells, and macrophages, in Helicobacter pylori�induced gastritis. Sixty-four patients (30 males, 34 females; mean age, 47 years) referred for upper endoscopy were recruited. Biopsy specimens from the gastric antrum and corpus were obtained for (1) detection of H. pylori and histological examination, (2) single and double immunostaining to test for expression of MIF protein in epithelial cells, T cells, and macrophages, and (2) in situ hybridization for expression of MIF mRNA within the lamina propria. In mucosal specimens from each of the 2 sites, both the percentage of MIF + epithelial cells and the numbers of MIF mRNA + inflammatory cells, MIF + T cells, and MIF + macrophages were significantly higher in H. pylori�positive patients than in H. pylori�negative patients. Overall, the percentage of MIF + epithelial cells and the numbers of MIF mRNA + cells, MIF + T cells, and MIF + macrophages were higher in the antrum than in the corpus. The percentage of MIF + epithelial cells and the numbers of MIF mRNA + cells, MIF + T cells, and MIF + macrophages increased in chronic gastritis, but, in the absence of H. pylori infection, this increase disappeared for all except MIF + T cells. Therefore, H. pylori infection is associated with increased expression of the MIF protein and MIF mRNA in gastric epithelial and inflammatory cells; along with other cytokines, MIF may play a significant role in gastric inflammation related to H. pylori infection.","author":[{"dropping-particle":"","family":"Xia","given":"Harry�Hua Xiang","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lam","given":"Shiu Kum","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Huang","given":"Xiao Ru","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Wong","given":"Wai Man","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Leung","given":"Suet Yi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Yuen","given":"Siu Tsan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lan","given":"Hui Yao","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Wong","given":"Benjamin�Chun Yu","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"The Journal of Infectious Diseases","id":"ITEM-1","issue":"2","issued":{"date-parts":[["2004"]]},"page":"293-302","title":"Helicobacter pylori Infection Is Associated with Increased Expression of Macrophage Migratory Inhibitory Factor�by Epithelial Cells, T Cells, and Macrophages�in Gastric Mucosa ","type":"article","volume":"190"},"uris":["http://www.mendeley.com/documents/?uuid=5596b908-f694-4dfc-ba66-871419b284da"]}],"mendeley":{"formattedCitation":"[7]","plainTextFormattedCitation":"[7]","previouslyFormattedCitation":"[7]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[7]. In their study Kebapcilar et al suggested that eradication of HP reduces levels of both MIF and hsCRP ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1590/s1807-59322010000800010","ISSN":"1807-5932","abstract":"OBJECTIVES: To determine the effect of Helicobacter pylori (H. pylori) eradication on blood levels of high-sensitivity C-reactive protein (hs-CRP), macrophage migration inhibitory factor and fetuin-A in patients with dyspepsia who are concurrently infected with H. pylori. METHODS: H.pylori infection was diagnosed based on the 14C urea breath test (UBT) and histology. Lansoprazole 30 mg twice daily, amoxicillin 1 g twice daily, and clarithromycin 500 mg twice daily were given to all infected patients for 14 days; 14C UBT was then re-measured. In 30 subjects, migration inhibitory factor, fetuin-A and hs-CRP levels were examined before and after the eradication of H. pylori infection and compared to levels in 30 healthy subjects who tested negative for H. pylori infection. RESULTS: Age and sex distribution were comparable between patients and controls. Migration inhibitory factor and hs-CRP levels were higher, and fetuin-A levels were lower, in H. pylori-infected patients (p<0.05). Following eradication of H. pylori, migration inhibitory factor and hs-CRP levels were significantly decreased, whereas fetuin-A levels were increased. However, eradication of the organism did not change lipid levels (p>0.05). CONCLUSION: These findings suggest that H. pylori eradication reduces the levels of pro-inflammatory cytokines such as migration inhibitory factor and hs-CRP and also results in a significant increase in anti-inflammatory markers such as fetuin-A.","author":[{"dropping-particle":"","family":"Kebapcilar","given":"Levent","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bilgir","given":"Oktay","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Cetinkaya","given":"Elif","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Akyol","given":"Murat","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bilgir","given":"Ferda","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bozkaya","given":"Giray","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Clinics","id":"ITEM-1","issue":"8","issued":{"date-parts":[["2010"]]},"page":"799-802","title":"The effect of Helicobacter pylori eradication on macrophage migration inhibitory factor, C-reactive protein and fetuin-a levels","type":"article","volume":"65"},"uris":["http://www.mendeley.com/documents/?uuid=d29240ee-c5f0-4175-b9c6-559883b7e66b"]}],"mendeley":{"formattedCitation":"[8]","plainTextFormattedCitation":"[8]","previouslyFormattedCitation":"[8]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[8]. 
Although both anti HP-IgG and IgA demonstrated high proportions in the sera of study groups, PCR results revealed no evidence of HP genome 16SrRNA. Similarly, Blasi et al showed no detection of bacterial DNA in peripheral blood  ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1086/315126","abstract":"Abdominal aortic aneurysm tissue and peripheral blood mononuclear cells (PBMC) of 41 consecutive subjects undergoing abdominal aortic aneurysm surgery were analyzed by polymerase chain reaction (PCR) for the presence of Chlamydia pneumoniae, Mycoplasma pneumoniae, and Helicobacter pylori DNA. Twenty patients (49%) were positive for C. pneumoniae DNA-16 (39%) in both PBMC and aneurysm tissue, 3 (7.3%) in PBMC only, and 1 (2.4%) in the artery specimen only. Previous exposure to C. pneumoniae was confirmed in 19 (95%) of the 20 PCR positive subjects by C. pneumoniae-specific serology, using the microimmunofluorescence test. None was positive for H. pylori or M. pneumoniae DNA, either in the PBMC or in the artery specimens. In conclusion, carriage of C. pneumoniae DNA is common both in PBMC and in abdominal aortic tissue from patients undergoing abdominal aneurysm surgery. Blood PCR may be a useful tool for identifying subjects carrying C. pneumoniae in the vascular wall.","author":[{"dropping-particle":"","family":"Blasi","given":"Francesco","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Boman","given":"Jens","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Esposito","given":"Gloria","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Melissano","given":"Germano","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Chiesa","given":"Roberto","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Cosentini","given":"Roberto","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Tarsia","given":"Paolo","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Tshomba","given":"Yamume","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Betti","given":"Monia","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Alessi","given":"Marinella","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Morelli","given":"Nicoletta","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Allegra","given":"Luigi","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"The Journal of Infectious Diseases","id":"ITEM-1","issue":"6","issued":{"date-parts":[["1999"]]},"page":"2074-2076","publisher":"Oxford University Press","title":"Chlamydia pneumoniae DNA Detection in Peripheral Blood Mononuclear Cells Is Predictive of Vascular Infection","type":"article","volume":"180"},"uris":["http://www.mendeley.com/documents/?uuid=e85a2ae1-57e5-40ab-a3b7-b066195e8e2e"]}],"mendeley":{"formattedCitation":"[17]","plainTextFormattedCitation":"[17]","previouslyFormattedCitation":"[17]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[17]. In contrary, Danesh et al. have detected HP�genomic DNA in buffy coat samples ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"ISSN":"00951137","author":[{"dropping-particle":"","family":"Danesh","given":"John","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Koreth","given":"John","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Youngman","given":"Linda","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Collins","given":"Rory","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Arnold","given":"J. Ranjit","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Balarajan","given":"Yarlini","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"McGee","given":"James","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Roskell","given":"Derek","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Journal of Clinical Microbiology","id":"ITEM-1","issue":"5","issued":{"date-parts":[["1999"]]},"page":"1651","title":"Is Helicobacter pylori a factor in coronary atherosclerosis?","type":"article-journal","volume":"37"},"uris":["http://www.mendeley.com/documents/?uuid=6c464ee1-2a4e-4457-a6a0-94fb1b3ca31a"]}],"mendeley":{"formattedCitation":"[18]","plainTextFormattedCitation":"[18]","previouslyFormattedCitation":"[18]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[18]
Such controversy results still observed over decades although plaque lesions were the specimen of choice for many study, for instance some studies reported positive results ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"ISSN":"00951137","author":[{"dropping-particle":"","family":"Danesh","given":"John","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Koreth","given":"John","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Youngman","given":"Linda","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Collins","given":"Rory","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Arnold","given":"J. Ranjit","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Balarajan","given":"Yarlini","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"McGee","given":"James","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Roskell","given":"Derek","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Journal of Clinical Microbiology","id":"ITEM-1","issue":"5","issued":{"date-parts":[["1999"]]},"page":"1651","title":"Is Helicobacter pylori a factor in coronary atherosclerosis?","type":"article-journal","volume":"37"},"uris":["http://www.mendeley.com/documents/?uuid=6c464ee1-2a4e-4457-a6a0-94fb1b3ca31a"]},{"id":"ITEM-2","itemData":{"ISSN":"00951137","abstract":"Chlamydia pneumoniae and Helicobacter pylori can cause persistent infections of the respiratory and gastrointestinal tract, respectively. It has been suggested that persistent infection of arteries with these bacteria can contribute to the development of atherosclerosis. The aims of this study were to determine the presence of C. pneumoniae and H. pylori DNA in atherosclerotic plaque samples by PCR and to evaluate the correlation between clinical status and DNA positivity of these bacteria. Eighty-five consecutive patients (mean age, 59 {+/-} 10; 75 male, 10 female) undergoing coronary artery bypass grafting, carotid endarterectomy, and surgery of the abdominal aorta for atherosclerotic obstructive lesions were included in the study. Forty-six endarterectomy specimens from the atherosclerotic lesions and 39 specimens from healthy regions of the ascending aorta, which were accepted as the control group, were excised. The presence of microorganism DNA in endarterectomy specimens was assessed by PCR. C. pneumoniae DNA was found in 12 (26%) of 46 endarterectomy specimens and none of the healthy vascular-wall specimens (P < 0.001), while H. pylori DNA was found in 17 (37%) of 46 endarterectomy specimens and none of the controls (P < 0.001). Either C. pneumoniae or H. pylori DNA was positive in 23 (50%) of 46 patients and none of the controls (P < 0.001). Six of the atherosclerotic lesions showed coexistence of both of the microorganism DNAs. The presence of C. pneumoniae and H. pylori DNA in a considerable number of atherosclerotic plaques but their absence in healthy vascular wall supports the idea that they may have a role in the development of atherosclerosis, especially in countries where infection is prevalent and where conventional risk factors fail to explain the high prevalence of atherosclerotic vascular disease.","author":[{"dropping-particle":"","family":"Farsak","given":"B.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Yildirir","given":"A.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Aky�n","given":"Y.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Pinar","given":"A.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"��","given":"M.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"B�ke","given":"E.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kes","given":"S.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Tokg�zo�lu","given":"L.","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Journal of Clinical Microbiology","id":"ITEM-2","issue":"12","issued":{"date-parts":[["2000"]]},"page":"4408-4411","title":"Detection of chlamydia pneumoniae and Helicobacter pylori DNA in human atherosclerotic plaques by PCR","type":"article","volume":"38"},"uris":["http://www.mendeley.com/documents/?uuid=1f88203d-4e1d-485f-b4b0-ea4b08d187fb"]},{"id":"ITEM-3","itemData":{"PMID":"18612444","abstract":"We investigated the relationship between acute coronary ischemia and the presence of Helicobacter pylori DNA in aortic regions that were absent macroscopic atheromatous plaques. The study group (Group 1) consisted of 42 patients who underwent coronary artery bypass grafting. Biopsy samples were obtained from 2 different locations: from regions of the aorta that were free (macroscopically) of atheromatous plaque (Group 1A), and from the internal mammary artery (Group 1B). The control group (Group 2) of 10 patients who had no atherosclerotic vascular disease provided aortic tissue samples for comparison. The real-time polymerase chain reaction method was used to detect H. pylori DNA in all biopsy samples. Eleven of 42 aortic tissue samples (26%) in Group 1A were positive for H. pylori DNA. Neither biopsies from the left internal mammary arteries of those patients nor biopsies from the aortas of the control group (Group 2) were positive for H. pylori DNA. There was a statistically significant difference between 1A and 1B in terms of H. pylori positivity (P=0.001). In Group 1 as a whole, acute coronary ischemia was more prevalent in the H. pylori-positive patients than in the H. pylori-negative patients (P=0.001). To our knowledge, this is the 1st study to investigate the detection of H. pylori DNA in aortic biopsy samples that are macroscopically free of atheromatous plaque. Such detection in patients who have atherosclerotic coronary artery disease could be an important indication of the role of microorganisms in the pathogenesis of atherosclerosis.","author":[{"dropping-particle":"","family":"Iriz E, Cirak MY, Engin ED, Zor MH, Erer D, Ozdogan ME, Turet S","given":"Yener A","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Texas Heart Institute Journal","id":"ITEM-3","issue":"2","issued":{"date-parts":[["2008"]]},"page":"130-5","title":"Detection of Helicobacter pylori DNA in aortic and left internal mammary artery biopsies","type":"article-journal","volume":"35"},"uris":["http://www.mendeley.com/documents/?uuid=5e663871-4421-4a17-a507-30c4a73c7153"]}],"mendeley":{"formattedCitation":"[18�20]","plainTextFormattedCitation":"[18�20]","previouslyFormattedCitation":"[18�20]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[18�20], while other did not despite of high HP seropositivity ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"ISSN":"00951137","abstract":"Recent reports suggest an association between Chlamydia pneumoniae and Helicobacter pylori bacteria and atherosclerosis. We studied 51 patients (mean age, 68.3 years) who underwent abdominal aortic aneurysm surgery. For each patient we performed a microimmunofluorescence test for immunoglobulin G (lgG), lgA, and lgM antibodies to C. pneumoniae specific antigen (TW-183). Anti-H. pylori antibodies were determined by means of an EIA-G test. Each aortic aneurysm surgical specimen was sampled into multiple sections of 0.3 cm 2 each and frozen at -21�C. Two samples of each aneurysm were used for a nested PCR with two sets of C. pneumoniae and two sets of H. pylori specific primers. Specimens were treated with a solution containing 20 mM Tris-HCl, Tween 20-Nonidet P-40 (0.5% [vol/vol each), and 100 �g of proteinase K per ml and incubated at 60�C for 1 h and at 98�C for 10 min. DNA was extracted twice with phenol-chloroform-isoamylic alcohol and precipitated with sodium acetate-ethanol by standard methods. Forty-one patients were seropositive for C. pneumoniae with past-infection patterns in 32 patients (16 d" IgG  <  512; 32 d" gA  <  256) and high antibody liters in 9 patients (IgG d" 512). In 26 of 51 patients, C. pneumoniae DNA was detected in aortic aneurysm plaque specimens. Of these patients, 23 had a serologic past-infection pattern, 2 had an acute reinfection pattern, and 1 was seronegative. Forty-seven of 51 patients were seropositive for H. pylori. In all cases PCR showed no evidence of H. pylori presence in plaque specimens. This study provides data on a possible C. pneumoniae involvement in the pathogenesis of aortic aneurysm and additional evidence for an association between this agent and atherosclerosis. Conversely, notwithstanding a high H. pylori seroprevalence observed, our results tend to rule not the possibility of a direct involvement of H. pylori in atherosclerosis.","author":[{"dropping-particle":"","family":"Blasi","given":"Francesco","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Denti","given":"Franco","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Erba","given":"Mario","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Cosentini","given":"Roberto","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Raccanelli","given":"Rita","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Rinaldi","given":"Angela","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fagetti","given":"Laura","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Esposito","given":"Gloria","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ruberti","given":"Ugo","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Allegra","given":"Luigi","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Journal of Clinical Microbiology","id":"ITEM-1","issue":"11","issued":{"date-parts":[["1996"]]},"page":"2766-2769","title":"Detection of Chlamydia pneumoniae but not Helicobacter pylori in atherosclerotic plaques of aortic aneurysms","type":"article","volume":"34"},"uris":["http://www.mendeley.com/documents/?uuid=282698ac-7d08-4824-b891-ba66d519f51b"]},{"id":"ITEM-2","itemData":{"DOI":"10.1016/j.carpath.2005.12.001","ISSN":"10548807","abstract":"Background and Introduction: It has been suggested that chronic infections may have a role in both the initiation and progression of atherosclerosis. While the majority of available data are focused on coronary artery disease, our aim was to investigate the presence of Chlamydia pneumoniae and Helicobacter pylori in samples from aortoiliac occlusive disease. Methods: Aorta-iliac atherectomy specimens were collected under sterile conditions from 21 patients (19 male, 2 female) undergoing surgery for aortoiliac occlusive disease. Seventeen macroscopically healthy vessels (12 internal mammary arteries, 3 radial arteries, prepared for coronary artery bypass graft, and 2 traumatic artery specimens, one of which was a superficial femoral artery and the other was a radial artery) were used as control. Blood samples for serological assays were obtained immediately before surgery. The polymerase chain reaction (PCR) was employed to search for H. pylori and C. pneumoniae DNA in atherosclerotic plaques and healthy vessel samples. Group-specific chlamydial lipopolysaccharide (LPS) antigens in atherosclerotic plaques and in healthy vessel samples and serum IgG antibodies to chlamydial LPS were determined by using a commercially available enzyme-linked immunosorbent assay (ELISA). Antibodies to H. pylori were also tested in all cases by means of an in-house ELISA. Results: Chlamydial LPS and DNA were detected in 6 of 21 (28.57%) atherosclerotic lesions using ELISA or PCR, respectively. There was no evidence of H. pylori DNA in any plaque specimens. All cases in which C. pneumoniae DNA was positive were also seropositive for antichlamydial LPS. Neither C. pneumoniae DNA nor antigen nor H. pylori DNA was found in the macroscopically healthy samples. Conclusion: Our results suggest that C. pneumoniae but not, as proposed, H. pylori may be involved in the pathogenesis of aortoiliac atherosclerosis. � 2006 Elsevier Inc. All rights reserved.","author":[{"dropping-particle":"","family":"Kaklikkaya","given":"Islam","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kaklikkaya","given":"Nese","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Buruk","given":"Kurtulu_","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Pulathan","given":"Zerrin","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Koramaz","given":"Ismail","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Aydin","given":"Faruk","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Tosun","given":"Ilknur","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kilic","given":"Ali Osman","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"�zcan","given":"Fahri","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Cardiovascular Pathology","id":"ITEM-2","issue":"2","issued":{"date-parts":[["2006"]]},"page":"105-109","title":"Investigation of Chlamydia pneumoniae DNA, chlamydial lipopolisaccharide antigens, and Helicobacter pylori DNA in atherosclerotic plaques of patients with aortoiliac occlusive disease","type":"article","volume":"15"},"uris":["http://www.mendeley.com/documents/?uuid=fe3505c8-380b-444f-970b-54893dbc0c85"]},{"id":"ITEM-3","itemData":{"DOI":"10.1023/A:1023059815117","ISSN":"01632116","abstract":"Chronic infections have been associated with cardiovascular disease. We used bacterial culture, polymerase chain reaction (PCR), and immunohistochemical staining with anti-vacA and anticagA antibodies to search for Helicobacter pylori and Chlamydiae pneumoniae in atherosclerotic plaques obtained at endarter ectomy. Serum IgG antibodies to H. pylori and C. pneumoniae were also determined. Thirty-two patients were enrolled. Anti-H. pylori and anti-C. pneumoniae IgG were present in 72% and 81%, respectively. Culture and PCR for H. pylori of vessel walls and plaques were negative. Atherosclerotic plaque and normal vessel sections from H. pylori-negative and- positive patients showed reactivity with anti-vacA and anti-cagA antibodies. C. pneumoniae DNA was amplified in three atherosclerotic lesions. These findings suggest that the association between H. pylori infection and atherosclerosis does not result from continuing direct effects of H. pylori antigens in the vessel walls. Antigens within vessel atherosclerotic plaques cross-react with H. pylori virulence factors and could act as cofactors in determining instability for the atherosclerotic plaques.","author":[{"dropping-particle":"","family":"Dore","given":"Maria Pina","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Sepulveda","given":"Antonia R.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bacciu","given":"Piero P.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Blasi","given":"Francesco","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Simula","given":"Luigi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Marras","given":"Lea","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Piccolo","given":"Davide","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Cherchi","given":"Giovanni B.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Graham","given":"David Y.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Realdi","given":"Giuseppe","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Digestive Diseases and Sciences","id":"ITEM-3","issue":"5","issued":{"date-parts":[["2003"]]},"page":"945-951","title":"Detection of Chlamydiae pneumoniae but not Helicobacter pylori DNA in atherosclerosis plaques","type":"article","volume":"48"},"uris":["http://www.mendeley.com/documents/?uuid=ca3ff9b7-7ed1-4145-8494-10c5805de6b0"]}],"mendeley":{"formattedCitation":"[21�23]","plainTextFormattedCitation":"[21�23]","previouslyFormattedCitation":"[21�23]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[21�23]. 
The insignificant difference between seropositive and negative MI group and insignificant correlation between MIF and anti HP-IgG assuming that HP may have a minor role in MIF elevation in MI. Thus, the origin of elevated MIF level may not attribute exclusively to HP infection as a whole source; however, cardiac origin appeared to be the major cause of increased MIF in MI since it has been revealed that MIF was produced intensively by different cells in all forms of atherosclerotic plaques, and appeared to have a significant role in the initiation of plaque lesions, and progression ADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1161/01.CIR.0000012942.49244.82","ISSN":"00097322","abstract":"Background - Atherosclerosis is a chronic inflammatory response of the arterial wall to injury. Macrophage migration inhibitory factor (MIF), a cytokine with potent inflammatory functions, was thus considered to be important in atherosclerotic lesion evolution. Methods and Results - We studied the presence and distribution of MIF immunoreactivity (MIF-IR) and MIF mRNA in internal mammary arteries with a normal histology and arteries with plaques in different stages of human atherosclerosis. To address a potential role for the coactivator Jab1 as a cellular mediator of MIF effects in vascular tissue, we correlated the expression of MIF to that of Jab1 by using immunohistochemistry and coimmunoprecipitation. We further sought to determine a potential functional role for endothelium-derived MIF in early atherogenesis by studying the effects of oxidized LDL on MIF expression in cultured human umbilical vascular endothelial cells. The results showed that MIF-IR and Jab1-IR are foun d in all cell types present in atherosclerotic lesions, that MIF-IR is upregulated during progression of atherosclerosis, that MIF is produced locally in the arterial wall, and that all MIF + cells are simultaneously Jab1 +. Coimmunoprecipitation experiments demonstrated in vivo complex formation between MIF and Jab1 in plaques. MIF expression in human umbilical vascular endothelial cells and a macrophage line was upregulated after stimulation with oxidized LDL. Conclusions - MIF is produced abundantly by various cells in all types of human atherosclerotic lesions and thus may play an important role in early plaque development and advanced complicated lesions. MIF-Jab1 complexes could serve critical regulatory functions in atherosclerotic lesion evolution.","author":[{"dropping-particle":"","family":"Burger-Kentischer","given":"Anke","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Goebel","given":"Heike","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Seiler","given":"R�diger","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fraedrich","given":"Gustav","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schaefer","given":"Hans E.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Dimmeler","given":"Stefanie","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kleemann","given":"Robert","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bernhagen","given":"J�rgen","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ihling","given":"Christian","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Circulation","id":"ITEM-1","issue":"13","issued":{"date-parts":[["2002"]]},"page":"1561-1566","title":"Expression of macrophage migration inhibitory factor in different stages of human atherosclerosis","type":"article","volume":"105"},"uris":["http://www.mendeley.com/documents/?uuid=bbfb4910-cae2-411d-a938-cd0b4dbd3660"]}],"mendeley":{"formattedCitation":"[24]","plainTextFormattedCitation":"[24]","previouslyFormattedCitation":"[26]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[24], verified by previously published studies which showed elevated MIF levels in CAD, and MI patientsADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1089/gtmb.2015.0113","ISSN":"1945-0265","abstract":"� Mary Ann Liebert, Inc. 2015. Background: Rupture of vulnerable plaque with subsequent thrombus formation has been implicated as the most common pathogenic mechanism responsible for acute coronary syndrome (ACS). Angiographic coronary lesion complexity has been reported to reflect plaque vulnerability. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine and might be involved in the pathophysiology of atherosclerotic plaque destabilization. Objective: This study was designed to investigate if serum MIF levels are associated with angiographic coronary lesion complexity in patients with coronary artery disease (CAD). Materials and Methods: A total of 232 consecutive CAD patients and 76 controls were recruited. CAD patients were subdivided according to the presence of ACS (n=138) or stable angina pectoris (SAP) (n=98). Coronary lesion morphology was assessed by coronary angiography. Serum MIF levels were measured by an enzyme-linked immunosorbent assay. Results: SAP patients had significantly higher serum MIF levels compared with healthy controls, and ACS patients had significantly higher serum MIF levels compared with SAP patients. In SAP patients, serum MIF levels were independently associated with the presence of complex coronary lesion. In ACS patients, serum MIF levels increased in conjunction with the extent of complex lesions. Conclusions: Serum MIF levels are a potential biomarker for reflecting the presence and severity of angiographically complex coronary lesion in CAD patients.","author":[{"dropping-particle":"","family":"Hao","given":"Yan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Yi","given":"Shao-Lei","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Zhong","given":"Jing-Quan","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Genetic Testing and Molecular Biomarkers","id":"ITEM-1","issue":"10","issued":{"date-parts":[["2015"]]},"page":"556-560","title":"Serum Macrophage Migration Inhibitory Factor Levels Are Associated with Angiographically Complex Coronary Lesions in Patients with Coronary Artery Disease","type":"article","volume":"19"},"uris":["http://www.mendeley.com/documents/?uuid=b2782b95-56d4-4976-8cea-f0bf563d757a"]},{"id":"ITEM-2","itemData":{"DOI":"10.1161/JAHA.113.000226","ISSN":"2047-9980","PMID":"24096574","abstract":"BACKGROUND: Early diagnosis and knowledge of infarct size is critical for the management of acute myocardial infarction (MI). We evaluated whether early elevated plasma level of macrophage migration inhibitory factor (MIF) is useful for these purposes in patients with ST-elevation MI (STEMI).\\n\\nMETHODS AND RESULTS: We first studied MIF level in plasma and the myocardium in mice and determined infarct size. MI for 15 or 60 minutes resulted in 2.5-fold increase over control values in plasma MIF levels while MIF content in the ischemic myocardium reduced by 50% and plasma MIF levels correlated with myocardium-at-risk and infarct size at both time-points (P < 0.01). In patients with STEMI, we obtained admission plasma samples and measured MIF, conventional troponins (TnI, TnT), high sensitive TnI (hsTnI), creatine kinase (CK), CK-MB, and myoglobin. Infarct size was assessed by cardiac magnetic resonance (CMR) imaging. Patients with chronic stable angina and healthy volunteers were studied as controls. Of 374 STEMI patients, 68% had elevated admission MIF levels above the highest value in healthy controls (> 41.6 ng/mL), a proportion similar to hsTnI (75%) and TnI (50%), but greater than other biomarkers studied (20% to 31%, all P < 0.05 versus MIF). Only admission MIF levels correlated with CMR-derived infarct size, ventricular volumes and ejection fraction (n = 42, r = 0.46 to 0.77, all P < 0.01) at 3 day and 3 months post-MI.\\n\\nCONCLUSION: Plasma MIF levels are elevated in a high proportion of STEMI patients at the first obtainable sample and these levels are predictive of final infarct size and the extent of cardiac remodeling.","author":[{"dropping-particle":"","family":"Chan","given":"William","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"White","given":"David A","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Wang","given":"Xin-Yu","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bai","given":"Ru-Feng","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Liu","given":"Yang","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Yu","given":"Hai-Yi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Zhang","given":"You-Yi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fan","given":"Fenling","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schneider","given":"Hans G","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Duffy","given":"Stephen J","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Taylor","given":"Andrew J","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Du","given":"Xiao-Jun","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Gao","given":"Wei","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Gao","given":"Xiao-Ming","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Dart","given":"Anthony M","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Journal of the American Heart Association","id":"ITEM-2","issue":"5","issued":{"date-parts":[["2013"]]},"page":"e000226","title":"Macrophage migration inhibitory factor for the early prediction of infarct size.","type":"article-journal","volume":"2"},"uris":["http://www.mendeley.com/documents/?uuid=5bb4270e-46b0-499a-bb92-b1544d6bda59"]}],"mendeley":{"formattedCitation":"[25,26]","plainTextFormattedCitation":"[25,26]","previouslyFormattedCitation":"[24,25]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[25,26], and MIF blockade result in plaque stabilization and regressionADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"10.1038/nm1567","ISSN":"10788956","abstract":"The cytokine macrophage migration inhibitory factor (MIF) plays a critical role in inflammatory diseases and atherogenesis. We identify the chemokine receptors CXCR2 and CXCR4 as functional receptors for MIF. MIF triggered G �i- and integrin-dependent arrest and chemotaxis of monocytes and T cells, rapid integrin activation and calcium influx through CXCR2 or CXCR4. MIF competed with cognate ligands for CXCR4 and CXCR2 binding, and directly bound to CXCR2. CXCR2 and CD74 formed a receptor complex, and monocyte arrest elicited by MIF in inflamed or atherosclerotic arteries involved both CXCR2 and CD74. In vivo, Mif deficiency impaired monocyte adhesion to the arterial wall in atherosclerosis-prone mice, and MIF-induced leukocyte recruitment required Il8rb (which encodes Cxcr2). Blockade of Mif but not of canonical ligands of Cxcr2 or Cxcr4 in mice with advanced atherosclerosis led to plaque regression and reduced monocyte and T-cell content in plaques. By activating both CXCR2 and CXCR4, MIF displays chemokine-like functions and acts as a major regulator of inflammatory cell recruitment and atherogenesis. Targeting MIF in individuals with manifest atherosclerosis can potentially be used to treat this condition. � 2007 Nature Publishing Group.","author":[{"dropping-particle":"","family":"Bernhagen","given":"J�rgen","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Krohn","given":"Regina","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Lue","given":"Hongqi","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Gregory","given":"Julia L.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Zernecke","given":"Alma","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Koenen","given":"Rory R.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Dewor","given":"Manfred","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Georgiev","given":"Ivan","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Schober","given":"Andreas","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Leng","given":"Lin","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kooistra","given":"Teake","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Fingerle-Rowson","given":"G�nter","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Ghezzi","given":"Pietro","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Kleemann","given":"Robert","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"McColl","given":"Shaun R.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Bucala","given":"Richard","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Hickey","given":"Michael J.","non-dropping-particle":"","parse-names":false,"suffix":""},{"dropping-particle":"","family":"Weber","given":"Christian","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Nature Medicine","id":"ITEM-1","issue":"5","issued":{"date-parts":[["2007"]]},"page":"587-596","title":"MIF is a noncognate ligand of CXC chemokine receptors in inflammatory and atherogenic cell recruitment","type":"article-journal","volume":"13"},"uris":["http://www.mendeley.com/documents/?uuid=2e7b373e-bd37-4ea2-bcdc-b8b529d53e42"]}],"mendeley":{"formattedCitation":"[27]","plainTextFormattedCitation":"[27]","previouslyFormattedCitation":"[27]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[27].
Clustering of most risk factors in MI seropositive HP group in which there was insignificant elevation of inflammatory biomarkers may revealed the indirect effects of the offending bacteria in disease progression through stimulating of pro-inflammatry, pro-coagulant, and pro-atherogenic environments, as infection with HP leads to prolonged chronic inflammatory reaction in the mucosa of gastric�tissue which may cause persistent release of low-level of systemic inflammatory mediators, therefore, increasing the risk of CADADDIN CSL_CITATION {"citationItems":[{"id":"ITEM-1","itemData":{"DOI":"https://doi.org/10.1016/0016-5085(92)90126-J","abstract":"Although Helicobacter pylori is now recognized as playing an etiologic role in chronic gastritis and peptic ulcer disease, information on the pathogenesis and natural history of infection is limited. A model is proposed in which luminal H. pyZori secrete substances that mediate inflammation that is beneficial to the organism but ultimately deleterious for the host; in addition to tissue damage, inflammation also affects gastric secretory function. In this model, the host may attempt to suppress the inflammatory response, and the adequacy of this postulated down-regulation determines pathological and clinical outcome. The effects of the inflammatory p
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����rocess on gastrin-hydrochloric acid homeostasis may be of critical importance in the pathogenesis of peptic ulcer disease. Because the long-term consequences of H. pylori colonization reflect the continued presence of the organism in the host over years or decades, it may be useful to consider this as a �slow� bacterial infection.","author":[{"dropping-particle":"","family":"Blaser","given":"M J","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"Gastroenterology","id":"ITEM-1","issue":"2","issued":{"date-parts":[["1992"]]},"page":"720-727","title":"Hypotheses on the pathogenesis and natural history of Helicobacter pylori-induced inflammation - ScienceDirect","type":"article-journal","volume":"102"},"uris":["http://www.mendeley.com/documents/?uuid=ac028386-f166-4bab-9a52-c9e29376a602"]},{"id":"ITEM-2","itemData":{"DOI":"http://dx.doi.org/10.1136/hrt.71.5.437","abstract":"Abstract Background-There is evidence suggesting that early life experience may influence adult risk of coronary heart disease (CHD). Chronic bacterial infections have been associated with CHD. Objective-To determine whether Helicobacter pylori, a childhood acquired chronic bacterial infection, is associated with an increased risk of coronary heart disease in later life. Design-Case-control study controlling for potential confounding variables with an opportunistically recruited control group. Subjects-111 consecutive cases with documented CHD were recruited from a cardiology clinic and 74 controls from a general practice health screening clinic. All were white men aged 45-65. Methods-Serum was analysed for the presence ofHpylori specific IgG antibodies by ELISA (98% sensitive and 94% specific for the presence of infection). Results-590/6 of the cases and 390/o of the controls were seropositive for H pylon (odds ratio 2*28, x2 7X35, p = 0.007). After adjustment by multiple logistic regression for age, cardiovascular risk factors, and current social class, the effect of H pylon was little altered (odds ratio 2X15, p = 0.03). Further adjustment for various features of the childhood environment known to be risk factors for H pylon infection only slightly weakened the association (odds ratio 1.9). Hpylori seropositivity was not related to the level of risk factors in the control population. Conclusion-In this pilot study the association of adult coronary heart disease with Hpylon seropositivity suggests that the early childhood environment may be important in determining the risk of CHD in adult life. The association needs confirmation in other better designed studies. IfHpyloni itself is responsible for the association, then this is of great potential importance as the infection is treatable.","author":[{"dropping-particle":"","family":"Mendall MA, Goggin PM, Molineaux M, Levy J, Toosy T, Strachan D, Camm AJ","given":"Northfield TC","non-dropping-particle":"","parse-names":false,"suffix":""}],"container-title":"British heart journal","id":"ITEM-2","issued":{"date-parts":[["1994"]]},"page":"437-439","title":"Relation of Helicobacter pylori infection and coronary heart disease","type":"article-journal","volume":"71"},"uris":["http://www.mendeley.com/documents/?uuid=e6d7e41f-3955-4894-b316-3fa2ab2fe630"]}],"mendeley":{"formattedCitation":"[28,29]","plainTextFormattedCitation":"[28,29]","previouslyFormattedCitation":"[28,29]"},"properties":{"noteIndex":0},"schema":"https://github.com/citation-style-language/schema/raw/master/csl-citation.json"}[28,29]. 


Conclusion 
Results of present study tend to exclude the possible direct contribution of HP in the pathogenesis of MI despite of high seroprevalence of anti HP-IgG, and suggest a possible indirect link between HP and low grade inflammatory reactions represented by insignificant elevation of MIF, and other inflammatory biomarkers which may lead to development of CAD and its complications such as MI. Further studies are on demand to support our findings.
Limitation: - Current study was done with a small sample size and initial demonstration of HP infection was just based on serology. Thus the study observations should be confirmed in a larger sample size and initial identification of bacterial infection should be confirmed by more than serological methods.
Funding: The author(s) received no specific funding for this work.
Conflicts of Interest: None.








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