Evaluation of MX1 Gene Promoter Methylation in Different Severities of COVID-19 Considering Patient Gender.

BACKGROUND: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), led to a pandemic in March 2020. During a viral infection, it has been reported that epigenetic changes occur for both sides: Infected cells elicit an antiviral environmental response, which induces and initiates certain pathways for proper response to the virus, while the virus silences the expression of vital genes in the anti-viral host cell. In this study, we aimed to examine the methylation level of the MX1 gene promoter in different stages in COVID-19 patients compared to the control group. METHODS: In total, 470 COVID-19 patients with a positive polymerase chain reaction (PCR) test (235 women and 235 men) were recruited into the study as the test group. Patients were divided based on the World Health Organization (WHO) classification into three groups: moderate, severe, and critical. Moreover, 100 healthy individuals (50 women and 50 men) were selected as the control group. Peripheral white blood cells were collected and PCR was performed using two types of primers designed for methylated and unmethylated states of the MX1 gene. The PCR products were then loaded on agarose gel and the band intensities were calculated by ImageJ software. RESULTS: The results showed a decrease in the methylation of the MX1 gene promoter in moderate and severe groups and an increase in the MX1 gene promoter methylation in the critical group. In addition, the level of methylation was higher in men than in women. CONCLUSIONS: Increased methylation of the MX1 gene in the critical group may indicate the role of SARS-CoV-2 in reducing the expression levels of this antiviral gene and thus promoting virus replication and disease progression.

Influence of Disease Severity and Gender on HLA-C Methylation in COVID-19 Patients.

In the pathophysiology of COVID-19, immunomodulatory factors play a vital role. Viruses have epigenetic effects on various genes, particularly methylation. Explaining the changes in immunological factor methylation levels during viral infections requires substantial consideration. HLA-C is a crucial determinant of immune function and NK cell activity and is primarily implicated in viral infections. This research focused on studying HLA-C methylation in COVID-19 patients with different severity. Peripheral blood samples were collected from 470 patients (235 men and 235 women) with RT-qPCR-confirmed COVID-19 test and classified into moderate, severe, and critical groups based on WHO criteria. Also, one hundred (50 men and 50 women) healthy subjects were selected as the control group. Peripheral blood mononuclear cells were used for DNA extraction, and the methylation-specific PCR (MSP) method and gel electrophoresis were used to determine the methylation status of the HLA-C. Significant statistical differences in HLA-C methylation were observed among cases and controls and various stages of the disease. HLA-C methylation in men and women has decreased in all stages (p < 0.05). In comparison with control, HLA-C methylation in both genders were as follows: moderate (women: 41.0%, men: 52.33%), severe (women: 43.42%, men: 64.86%), critical (women: 42.33%, men: 60.07%), and total patients (women: 45.52%, men: 56.97%). Furthermore, the methylation levels in men were higher than in women in all groups (p < 0.05). Significantly, among all groups, the severe group of men participants showed the highest methylation percentage (p < 0.05). No significant differences were detected for different disease severity in the women group (p > 0.1). This study found that HLA-C methylation was significantly lower in COVID-19 patients with different disease severity. There were also significant differences in HLA-C methylation between men and women patients with different severity. Therefore, during managing viral infections, particularly COVID-19, it is critical to consider patient gender and disease severity.

CCR2 and DPP9 expression in the peripheral blood of COVID-19 patients: Influences of the disease severity and gender.

INTRODUCTION: Hyper-inflammatory reactions play a crucial role in the pathogenesis of the severe forms of COVID-19. However, clarification of the molecular basis of the inflammatory-related factors needs more consideration. The aim was to evaluate the gene expression of two fundamental molecules contributing to the induction of inflammatory like CCR2 and DPP9 in cells from peripheral blood samples from patients with various patterns of COVID-19. METHODS: Peripheral blood samples were collected from 470 patients (235 male and 235 female) with RT-qPCR-confirmed COVID-19 test exhibiting moderate, severe, and critical symptoms based on WHO criteria. 100 healthy subjects (50 male and 50 female) were also enrolled in the study as a control group. The gene expression of DPP-9 and CCR-2 was assessed in the blood samples using real-time PCR method. RESULTS: The COVID-19 patients in severe stage expressed higher levels of CCR2 and DPP9 compared with healthy controls. In male and female patients, the levels of CCR2 and DDP9 expression significantly differed between moderate, severe, and critical patterns (p < 0.0001) as well as between each COVID-19 form and control group (p < 0.0001). The male patients with severe COVID-19 expressed greater levels of CCR2 and DPP-9 than female with same disease form. The female patients with moderate and critical COVID-19 expressed greater levels of CCR2 and DPP-9 than male patients with same disease stage. CONCLUSION: We demonstrated that the expression of DPP-9 and CCR-2 was substantially increased in COVID-19 patients with different forms of disease. Considerable differences were also demonstrated between male and female with different patterns of disease. Therefore, we suggest to consider the gender of patients and disease severity for management of COVID-19.

The optimal diagnostic strategies for patient with coronary artery diseases and stable chest pain syndrome: a cost-effectiveness analysis.

BACKGROUND: Numerous invasive and noninvasive diagnostic tests with different cost and effectiveness exist for detection of coronary artery disease. This diversity leads to unnecessary utilization of health services. For this reason, this study focused on the cost-effectiveness analysis of diagnostic strategies for coronary artery disease from the perspective of the health care system with 1-year time horizon. RESULTS: Incremental cost effectiveness ratios of all strategies were less than the threshold except for the electrocardiography-computed tomography angiography-coronary angiography strategy, and cost of the cardiac magnetic resonance imaging-based strategy was higher than the cost of other strategies. Also, the number of correct diagnosis in the electrocardiography-coronary angiography strategy was higher than the other strategies, and its ICER was 15.197 dollars per additional correct diagnosis. Moreover, the sensitivity analysis found that the probability of doing MRI and sensitivity of the exercise electrocardiography had impact on the results. CONCLUSION: The most cost-effective strategy for acute patient is ECG-CA strategy, and for chronic patient, the most cost-effective strategies are electrocardiography-single photon emission computed tomography-coronary angiography and electrocardiography-exercise electrocardiography-coronary angiography. Applying these strategies in the same clinical settings may lead to a better utilization of resources.

A Successful implementation of an idea to a nationally approved plan: Analyzing Iran's National Health Roadmap using the Kingdon model of policymaking.

Introduction: Hospital beds, human resources, and medical equipment are the costliest elements in the health system and play an essential role at the time of treatment. In this paper, different phases of the NEDA 2026 project and its methodological approach were presented and its formulation process was analysed using the Kingdon model of policymaking. Methods: Iran Health Roadmap (NEDA 2026) project started in March 2016 and ended in March 2017. The main components of this project were hospital beds, clinical human resources, specialist personnel, capital medical equipment, laboratory facilities, emergency services, and service delivery model. Kingdon model of policymaking was used to evaluate NEDA 2026 development and implementation. In this study, all activities to accomplish each step in the Kingdon model was described. Results: The followings were done to accomplish the goals of each step: collecting experts' viewpoint (problem identification and definition), systematic review of the literature, analysis of previous experiences, stakeholder analysis, economic analysis, and feasibility study (solution appropriateness analysis), three-round Delphi survey (policy survey and scrutinization), and intersectoral and interasectoral agreement (policy legislation). Conclusion: In the provision of an efficient health service, various components affect each other and the desired outcome, so they need to be considered as parts of an integrated system in developing a roadmap for the health system. Thus, this study demonstrated the cooperation process at different levels of Iran's health system to formulate a roadmap to provide the necessary resources for the health sector for the next 10 years and to ensure its feasibility using the Kingdon policy framework.

Relationship between serum N-terminal Pro Brain Natriuretic Peptide (NT-Pro BNP) level and the severity of coronary artery involvements.

BACKGROUND: Rapid measuring of B-type natriuretic peptide (BNP) in the emergency departments effectively results in evaluating patients with acute cardiac attacks and has appeared to be a useful prognostic marker of cardiovascular risk. A current study came to address the association between plasma N-terminal pro BNP level and severity of coronary vessels' defects based on Gensini score in patients with stable angina pectoris candidate for coronary angiography. METHODS: The study population consisted of 92 consecutive patients with appearance of stable angina and candidate for coronary angiography. All participants underwent selective left and right coronary angiography. For BNP measurement and just before the catheterization of left coronary, 5cc blood samples were drawn from coronary. RESULTS: With respect to the role of N terminal pro BNP for predicting severity of CAD based on Gensini scoring, linear regression analysis confirmed that plasma BNP level was a strong predictor for CAD severity (p = 0.009) in the presence of study cofounders. A significant correlation was also observed between N terminal pro BNP and left ventricular ejection fraction, so that all patients with left ventricular dysfunction (EF < 40%) had plasma N terminal pro BNP level higher than 100 pg/ml. CONCLUSIONS: NT-pro BNP can be a good parameter for predicting the severity of coronary vessels' involvement besides other diagnostic tools. In all patients with left ventricular ejection fraction less than 40%, plasma NT-pro BNP level was higher than 100 pg/ml.

Diagnostic performance of electrocardiography in the assessment of significant coronary artery disease and its anatomical size in comparison with coronary angiography.

BACKGROUND: Current study addressed the predictive value of 12-lead electrocardiogram (ECG) in patients with suspected coronary artery disease (CAD). METHODS: Four hundred consecutive patients with new onset of chest pain were studied. A resting standard 12-lead ECG was recorded and all patients underwent coronary angiography. RESULTS: ECG correctly detected significant stenosis in 176 out of 400 patients with an overall sensitivity per patient of 51.5% and specificity per patient of 66.1%. Based on artery analysis, ECG had the highest and lowest sensitivity for the detection of involvement in LAD (37.3%) and RCA (25.8%), respectively. ROC curve analysis showed that ECG changes were not good indicators of coronary arteries involvement with areas under the ROC curves 0.586 (for LAD artery), 0.524 (for RCA artery) and 0.530 (for LCX artery). CONCLUSIONS: ECG has low partial sensitivity and specificity for predicting coronary artery stenosis with accuracy ranged 58.5 to 62.0 percent based on coronary artery analysis.

Survey of changes in complete blood count and red cell indices of whole blood incubated in vitro at different temperatures up to 48 hours.

BACKGROUND: Complete blood count (CBC) is one of the most common and conventional blood test that physicians usually request. However the results of this test are affected by different factors such as, the temperature and duration of incubation, therefore the aim of this survey was to evaluate the effect of temperature and time of incubation on CBC, red blood cells (RBC) indices and white blood cells (WBC) differential count. METHODS: In a cross-sectional study, blood samples were taken from 30 healthy medical students of Rafsanjan University (15 males and 15 females). The samples divided into three parts; CBC were done on the samples up to 48 hours incubation at temperature of 25, 30, and 370 C at the time of sampling, and after 2, 8, 24 and 48 hours. Data were statistically analyzed and the following results were obtained. RESULTS: RBC count, hematocrit, MCH, percent of monocytes and eosinophils were constant in different temperatures, WBC count, MCHC, hemoglobin, platelets count, the percent of lymphocytes and neutrophils were constant up to 24 hours and then tend to increase with increasing temperature except lymphocytes percent that tend to decrease. MCV decreased with increasing temperature up to 8 hours and then significantly increased (from 83.89 to 87.50 fmol/l, p < 0.001). WBC, hematocrit, MCV, platelets count, and neutrophils' percent tend to increase by the time of incubation, but RBC count, MCHC, lymphocytes' percent decreased. Hemoglobin, MCH, and the percent of monocytes and eosinophils were constant. CONCLUSION: The finding of this survey showed that some of CBC parameters can be changed with the incubation, therefore it is better to do the CBC test after blood taking as soon as possible.