Development of an effective liposomal cholesterol ester transfer protein (CETP) vaccine for protecting against atherosclerosis in rabbit model.

Clinical trials of cholesterol ester transfer protein (CETP) peptide vaccine were stopped after disappointing results in humans due to the inadequacy of adjuvant aluminum hydroxide in stimulating the immune response against the self-antigen of CETP. To increase the efficacy of the CETP vaccine, we developed a novel liposomal form of tetanus toxoid-CETP (TT-CETP) peptide (Lip CETP) with well-characterized properties and high encapsulation efficiency. The vaccine efficacy against atherosclerosis was evaluated in rabbits challenged with a high cholesterol diet. Rabbits were immunized with Lip-CETP or liposome containing CETP with CpG ODN (Lip CETP/CpG). Control groups received empty liposomes or buffer. Anti-TT-CETP specific antibodies in serum were determined and gene expression of cytokine IFN-γ and IL-4 were measured in blood peripheral mononuclear cells. Therapeutic response was evaluated by titration of plasma lipoproteins during the study and pathologic analysis of aorta atherosclerotic lesions at the end. Lip-CETP/CpG elicited strong anti-TT-CETP antibodies and a higher IFN-γ level than the buffer. IL-4 was lower than the buffer in all vaccinated groups. Plasma lipoproteins showed no significant difference in the studied groups. Atherosclerosis thickness grade of the aorta was lower than the buffer group (p < 0.001) in rabbits vaccinated with Lip-CETP but not with Lip-CETP/CpG. In conclusion, Lip-CETP showed a strong atheroprotective effect.

Modeling of chromium (VI) removal from aqueous solution using modified green-Graphene: RSM-CCD approach, optimization, isotherm, and kinetic studies.

BACKGROUND: The aim of this study was to investigate the removal of Cr (VI) using Green-Graphene Nanosheets (GGN) synthesized from rice straw. METHODS: Synthesis of the GGN was optimized using response surface methodology and central composite design (CCD). The effect of two independent variables including KOH-to-raw rice ash (KOH/RRA) ratio and temperature on the specific surface area of the GGN was determined. To have better removal of Cr (VI), GGN was modified using the grafting amine group method. In the Cr (VI) removal process, the effects of four independent variables including initial Cr (VI) concentration, adsorbent dosage, contact time, and initial solution pH were studied. RESULTS: The results of this study showed that the optimum values of the KOH/RRA ratio and temperature for the preparation of GGN were 10.85 and 749.61 °C, respectively. The maximum amount of SSA obtained at optimum conditions for GGN was 551.14 ± 3.83 m 2 /g. The optimum conditions for Cr (VI) removal were 48.35 mg/L, 1.46 g/L, 44.30 min, and 6.87 for Cr (VI) concentration, adsorbent dosage, contact time, and pH, respectively. Based on variance analysis, the adsorbent dose was the most sensitive factor for Cr (VI) removal. Langmuir isotherm (R2 = 0.991) and Pseudo-second-order kinetic models (R2 = 0.999) were the best fit for the study results and the Q max was 138.89 mg/g. CONCLUSIONS: It can be concluded that the predicted conditions from the GGN synthesis model and the optimum conditions from the Cr (VI) removal model both agreed with the experimental findings.

Could decorin be a biomarker of coronary artery disease? A pilot study in human beings.

BACKGROUND AND AIM: Nowadays there is a strong necessity in identifying patients who may be exposed to the risk for future cardiovascular events like progressive atherosclerotic disease. Biomarkers are valuable tools for this purpose. Coronary artery calcification (CAC) is utilized as an important tool for the global risk assessment of cardiovascular events in individuals with intermediate risk. Decorin (DCN) is a small leucine-rich proteoglycan that induces calcification of arterial smooth muscle cell and localizes to mineral deposition in human atherosclerotic plaque. The main purpose of this clinical study was to find out the correlation between Decorin serum concentration and CAC in human for the first time. METHODS: In this study 84 patients with coronary artery disease who fulfilled inclusion and exclusion criteria, entered the study. For all patients a questionnaire consisting demographic data and traditional cardiovascular risk factors were completed. CT-Angiography was carried out to determine coronary artery calcium score and ELISA method was used for measuring DCN serum concentrations. RESULTS: No significant correlation between DCN serum concentration and total CAC score and also CAC of left anterior descending, right coronary artery, left main coronary artery and circumflex was found in the study population (P>0.05). CONCLUSIONS: On the basis of our results DCN serum concentration is not a suitable biomarker of coronary artery disease. However, more studies with higher sample size are necessary for its confirmation.

Evaluation of NPP1 as a Novel Biomarker of Coronary Artery Disease: A Pilot Study in Human Beings.

Purpose: Coronary artery calcification (CAC) is utilized as an important tool for global risk assessment of cardiovascular events in individuals with intermediate risk. Ecto phosphodiesterase/nucleotide phosphohydrolase-1(ENPP1) converts extracellular nucleotides into inorganic pyrophosphate and it is a key regulator of tissue calcification that adjusts calcification in tissues like vascular smooth muscle cells. The main purpose of this clinical study was to find out the correlation between ENPP1 serum concentration and CAC in human for the first time. Methods: In this study 83 patients (16 diabetic patients and 67 non-diabetic patients) with coronary artery disease who fulfilled inclusion and exclusion criteria, entered the study. For all patients a questionnaire consisting demographic data and traditional cardiovascular risk factors were completed. Computed tomography (CT)-Angiography was carried out to determine coronary artery calcium score and enzyme-linked immunosorbent assay (ELISA) method was used for measuring ENPP1 serum concentrations. Results: There was a reverse significant correlation between ENPP1 serum concentration and total CAC score and also CAC of right coronary artery (RCA) (P<0.05) in non-diabetic patients. Conclusion: On the basis of our results, ENPP1 serum concentration may be a suitable biomarker for coronary artery disease at least in non-diabetic patients. However, more studies with higher sample size are necessary for its confirmation.

Anti-atherosclerosis effect of different doses of CETP vaccine in rabbit model of atherosclerosis.

AIM: To evaluate atheroprotective effects of different doses of cholesteryl ester transfer protein (CETP) vaccine, three doses of Tetanus toxoid-CETP (TT-CETP) peptide including 10, 50 and 100/rabbit, termed FA10, FA50, FA100, respectively, were administered in rabbit model of atherosclerosis. METHODS: Animals were vaccinated subcutaneously (S.C.) with 100µl of vaccine in presence of complete Freund's adjuvant (CFA) for the first administration. Rabbits were boosted 4 times at 3 weeks intervals with the same peptide dose formulated in incomplete Freund's adjuvant (IFA). Animals were fed with diet supplemented with 2% cholesterol from week 11 to week 19. Anti-TT-CETP specific antibody and CETP activity in sera were determined. Therapeutic response was examined by tracking plasma lipoprotein levels (HDL-C, LDL-C and total cholesterol), and pathologic observation of intima/media thickness at the site of aortic lesions. RESULTS: All TT-CETP vaccine doses generated strong anti TT-CETP antibody response. CETP activity reduced in rabbits vaccinated with FA100 (P=0.031). FA100 showed significant increase in level of HDL-C rather than control group (P=0.006). However, no significant reduction were found in atherosclerotic lesion when compared to control. CONCLUSION: Inhibition of CETP activity and increased HDL-C were found with FA100, but the vaccine failed to prevent aortic lesion development in immunized rabbits when compared to control. Our result supports the hypothesis stated that CETP may not be an attractive therapeutic target for the prevention of cardiovascular disease.

A novel atheroprotective role of MF59-like adjuvant when co-administered with CETP vaccine in rabbit model of atherosclerosis.

OBJECTIVES: In this study, for the first time, MF59 adjuvant was used to develop a cholesteryl ester transfer protein (CETP) vaccine. The efficacy of the vaccine was compared with the efficacy of CETP vaccine formulated with Alum/CpG, the formulation that its immunogenicity has been already demonstrated in rabbit and mice. MATERIALS AND METHODS: Tetanus toxoid- CETP peptide (TT-CETP) was mixed with Alum/CpG or MF59-like and administered subcutaneously for total five times in rabbit model of atherosclerosis. Anti-TT-CETP specific antibody, CETP activity in sera and mRNA level of cytokine IL-4 and IFN-γ in peripheral mononuclear cells were determined. Therapeutic response was also examined by tracking serum lipoprotein levels and pathologic observation of atherosclerotic lesions at aortic site. RESULTS: More anti-TT-CETP antibody was found in Alum/CpG vaccinated rabbits compared to buffer (P<0.001). Antibody induced by MF59-like formulation was not significantly higher than buffer. CETP activity and lipoprotein levels were not significantly different between vaccinated and control rabbits. The mRNA level of IL-4 was significantly lower than buffer while, IFN-γ gene expression was significantly higher in both vaccinated groups. Atherosclerosis thickness grade of aorta was dramatically lower than buffer (P<0.01) in both vaccinated groups. CONCLUSION: It is concluded that MF59-adjuvanted CETP vaccine showed anti-atherosclerosis properties, but the protective effect could not be directly attributed to the immune response induced by anti TT-CETP antibody and CETP inhibition. Further studies are needed to explain the anti-atherosclerosis properties of MF59 in the presence of TT-CETP peptide.

Smoking and Its Related Factors Among Iranian High School Students.

BACKGROUND: In different studies, the prevalence of tobacco consumption has been growing in high schools boys. OBJECTIVES: This study was conducted to determine the prevalence of smoking and its related factors among Iranian high school students in 2011. MATERIALS AND METHODS: In this cross-sectional study, 450 male students from 15 high schools of Shahroud (northeast of Iran) were selected for evaluation of the knowledge, attitude and practices (KAP) of students regarding tobacco consumption. RESULTS: Overall, 51% (95% CI: 46.5 - 55.7) of the students had positive history of smoking for at least one time and 7.1% (95% CI: 5 - 10) of them were current smokers. The most prevalent source of information about smoking was TV and radio programs (48%) and friends were the second source (22%). Based on the students' opinions, entertainment and smoker friends were the most important reasons for smoking tendency. There was significant statistical association between students smoking and positive family history of smoking (P value < 0.05). CONCLUSIONS: The prevalence of smoking experince was very high among high school students. The most prevalent source of information about smoking was Iranian broadcasting companies. Positive family history of smoking and smoker friends were the important motivating factors toward smoking.

Simple screening of pulmonary artery hypertension using standard chest x ray: an old technique, new landmark.

BACKGROUND: Pulmonary artery hypertension (PAH) is difficult to diagnose because of its nonspecific symptoms. Although echocardiography can reliably and rapidly recognize the presence of pulmonary hypertension, chest X ray (CXR) is more widely used because of its availability. The objective of this study was to find a parameter, which, by changing the scale of CXR, is still useful for detection of PAH. MATERIALS AND METHODS: This case control prospective study included 100 subjects with a clinical finding of dyspnea. Additionally, thirty healthy volunteers (control group) were included in this study. Systolic pulmonary artery pressure (SPAP) was determined by echocardiography. Widening of pulmonary hilum, projection of the right side of the heart border (PRHB) and the ratio of these parameters to the chest diameter were compared to SPAP using the regression method. A cut-off point was determined for parameters that showed significant correlation. RESULTS: The most prevalent disease in the dyspnea group was COPD (28%). Average SPAP was 41.8±17.3 (ranging from 10 to 87 mmHg). Multivariate analysis of the covariance revealed significant correlation between SPAP, age, sex and hilar widening (r = 0.44, P = 0.0001) that was higher than PRHB and hilar widening + PRHB (r= 0.374. and r= 0.438, respectively). The ROC curve showed that the area under the curve was not significantly different for all parameters and the best cut-off point with sensitivity of more than 80% was as follows: hilar size more than 112 mm, PRHB more than 44 and hilum/chest ratio more than 0.44. CONCLUSION: Hilum/chest ratio is the proper substitution for the hilar size in case of changing the scale of the chest X ray. Evaluation of hilar widening and PRHB could lead to identifying more subjects suffering from undiagnosed PAH.

Evaluation of RANKL/OPG Serum Concentration Ratio as a New Biomarker for Coronary Artery Calcification: A Pilot Study.

Objective. There is a strong need for biomarkers to identify patients at risk for future cardiovascular events related with progressive atherosclerotic disease. Osteoprotegerin (OPG) protects the skeleton from excessive bone resorption by binding to receptor activator of nuclear factor-κB ligand (RANKL) and preventing it from binding to its receptor, receptor activator of nuclear factor-κB. However, conflicting results have been obtained about association of serum level of OPG or RANKL with coronary artery disease (CAD). Based on their role in inflammation and matrix degradation and the fact that atherosclerotic plaque formation is an inflammatory process, we hypothesized that RANKL : OPG ratio could be a better biomarker for CAD. Methods. In this cross-sectional study, the correlation between RANKL : OPG ratio serum concentration and coronary artery calcification (CAC) in 50 patients with ischemic coronary disease has been investigated. We used ELISA method for measuring RANKL and OPG serum concentrations. Results. There was a significant correlation between RANKL : OPG serum concentration ratio and CAC in our study population (P = 0.01). Conclusion. Our results suggested that RANKL : OPG ratio concentration has a potential of being used as a marker for coronary artery disease.