Methods: Serum LDL and HDL subclasses are separated by native gradient gel electrophoresis (4–12%, Biorad, Hercules, CA), which is followed by transfer and sequential western blot to detect apolipoproteins AI, AII, B, E, CII and CIII and lp(a). In this way distinct apolipoprotein profiles across HDL subclasses are obtained. In a pilot study we tested our method on purified HDL and LDL as well as in serum from hyperlipidemic patients and gender and age-matched healthy controls. HDL subclasses are also analyzed for comparison and control purposes using the Lipoprint HDL system from Quantimetrix (Redondo Beach, CA). Results and conclusions: Our method allows for detection of the major apo- lipoproteins in native HDL subclasses and shows a striking differential dis- tribution in different subjects, independent of the HDL-C. Differences in sdLDL, apoE, CII and CIII are striking when hyperlipidemic and control sub- jects are compared. Our procedure is more practical for routine use than MALDI-MS proteomics on purified HDL by ultracentrifugation, which also has the disadvantage of high ionic strengths that lead to loss of surface proteins. POMEGRANATE JUICE (PJ) PREVENTS ENDOTHELIAL DYSFUNCTION INDUCED BY PRIMED NEUTROPHILS Ricardo Almuly a , Batya Kristal b, c , Eynav kliger a, c , Boaz Elad a, c , Galina Shapiro a , Shifra Sela a,c a Eliachar Research Laboratory, Western Galilee Medical Center, Nahariya, Israel; b Department of Nephrology and Hypertension, Western Galilee Medical Center, Nahariya, Israel; c Faculty of medicine in the Galilee, Bar- Ilan University, Zefat, Israel Background: Primed neutrophils are mediators of oxidative stress (OS) and inflammation, key contributors to the pathogenesis of endothelial dysfunction underlying atherosclerosis and cardiovascular (CV) diseases. We have reported that one year of pomegranate juice (PJ) intake by he- modialysis (HD) patients attenuated the atherosclerotic process and reduced traditional and non-traditional CV risk factors, including reduc- tion in neutrophils priming. However, standard formulation and/or mechanisms explaining the beneficial PJ actions are still needed. Methods: We developed a co-culture system which includes endothelial cells, neutrophils and monocytes in order to elucidate possible beneficial pathways of PJ action. Endothelial cells were pretreated with 1% PJ before exposure to H 2 O 2 or to neutrophils separated from HD patients. The expression levels of endothelial oxidative and inflammatory markers and monocytes adhesion and transmigration were studied. Results: In this study PJ action as a direct scavenger was excluded, since PJ and oxidizing agents were never in contact. Pretreatment of endothelial cells with 0.5 – 1% PJ before exposure to H 2 O 2 resulted in improved cell survival, lowered over-expression of heme oxygenase (HO-1) and IL-6 and attenuated the reduction in eNOS and Sirt-1 gene expression. PJ reduced the adhesion of monocytes to endothelial cells induced by either TNF-a or neutrophils from HD patients. Conclusions: PJ improves cell survival and confers anti-inflammatory and anti-oxidative effects on endothelial cells, partly explaining the results of our one-year clinical study, showing attenuation of the atherosclerotic process by PJ. Further studies are on their way to standardize PJ active constituents for future standardization as food supplement. ACCELERATED ATHEROSCLEROSIS IN INDIVIDUALS WITH THE HAPTOGLOBIN 2-2 GENOTYPE AND DIABETES IS MEDIATED BY MACROPHAGE LYSOSOMAL INJURY AND APOPTOSIS Rabea Asleh a, b , John Ward a , Nina S. Levy a , Shady Safuri a , Doron Aronson b , Andrew P. Levy a a Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel; b Department of Cardiology, Rambam Health Care Center, Haifa, Israel Background: There exists a common functional allelic polymorphism in the Haptoglobin (Hp) gene which has been associated with the risk of cardio- vascular disease in individuals with diabetes mellitus (DM). In the athero- sclerotic plaque, the CD163 macrophage Hp-hemoglobin (Hb) receptor is responsible for clearing Hb released after intraplaque hemorrhage. In DM, the Hp 2 allele protein product is defective in its Hb scavenging properties and has been associated with increased iron in the atherosclerotic plaque. Hypothesis: We proposed that the increased iron accumulation in Hp 2-2 plaques and DM was due to an impaired processing of the Hp 2-2-Hb complex within macrophage lysosomes and that this iron was toxic to the macrophage lysosomal membrane thereby resulting in the loss of lyso- somal membrane integrity and cellular injury. Methods and results: Confocal microscopy using lysotropic pH indicator dyes was used to demonstrate that uptake of Hp 2-2-Hb complexes by the macrophage CD163 receptor disrupted the lysosomal pH gradient. Cellular fractionation studies of lysosomes isolated from macrophages incubated with Hp 2-2-Hb complexes demonstrated increased lysosomal membrane oxida- tion and a loss of lysosomal membrane integrity. We found that the lysosomal acidic pH was an essential component responsible for redox active iron generation and iron induced lysosomal oxidative injury associated with Hp 2- 2-Hb uptake. Additionally, several apoptotic markers (including DNA frag- mentation and active caspase-3) were significantly increased in macrophages endocytosing the Hp 2-2-Hb complexes. The oxidative lysosomal injury and macrophage apoptosis associated with Hp 2-2-Hb complexes were dramat- ically enhanced when using glycosylated Hp-Hb complexes. Conclusions: Lysosomal injury and subsequent macrophage apoptosis secondary to uptake of Hp 2-2-Hb complexes may play a major role in the development of diabetic atherosclerosis. SCREENING FOR DIABETES AMONG IN-PATIENTS ADMITTED TO INTERNAL MEDICINE DEPARTMENTS – QUALITY CONTROL SURVEY Eyal Leibovitz, Mona Boaz, Inna Baer, Julio Wainstein The Department of Internal Medicine “E”, The Epidemiology Unit and Diabetes Clinic at the Wolfson Medical Center, Holon, and the Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel Aim: To evaluate the attention of hospital physicians, as expressed in discharge recommendations, to glucose levels above 200 mg/dL in the blood of patients without the diagnosis of diabetes mellitus. Methods: This is a retrospective analysis of patient records. Patients were considered as having pathological random sample glucose (RSG) if one of the first 2 blood glucose levels were above 200 mg%. For patients without known diabetes mellitus, the discharge letters were audited to evaluate the specific recommendations issued. Results: Included were 5274 patients (mean age of 68.2  18.2 years, and 57.3% were males, 28.0% diabetes). Among the 3795 patients without diabetes upon admission, 211 patients (5.4%) had pathological RSG. Compared to the non-pathological RSG, these patients were older (75.8  16.0 Vs. 66.5  19.7 years, p < 0.001), with a higher incidence of renal disease (14.37% Vs. 8.8%, p ¼ 0.003), and malignancy (17.5% Vs, 11.3%, p ¼ 0.006). Of the 211 patients pathological RSG, 156 patients (73.9%) were discharged home or to a nursing home. Among this sub-population, 84% had no recommendation regarding the pathological blood test, 10.3% had a recommendation to start anti-diabetes medications, 4.4% were recom- mended to repeat the lab tests and 1.3% were recommended to change the lifestyle habits. Patients without a recommendation were older and had higher prevalence of mental illness. Conclusion: Elevated random sample glucose above 200 mg/dL is found in 5.4% of non-diabetes patients. Despite the high frequency, more than 80% of them had no recommendation for diabetes screening or treatment in the discharge letter. ASSOCIATION BETWEEN GLUCOSE CONTROL DURING ADMISSION AND IN-HOSPITAL PROGNOSIS AMONG DIABETES PATIENTS ADMITTED TO INTERNAL MEDICINE DEPARTMENTS Massarawa Muhamed, Mona Boaz, Julio Wainstein, Eyal Leibovitz The Department of Internal Medicine “E”, The Epidemiology Unit and Diabetes Clinic at the Wolfson Medical Center, Holon, and the Sackler school of Medicine, Tel Aviv university, Tel Aviv, Israel Abstracts / Atherosclerosis 233 (2014) 326–330 328