MS490 LARGE-SCALE PHENOTYPING OF EMP3-KNOCKOUT MICE AT THE GERMAN MOUSE CLINIC REVEALS A SPECIFIC ROLE OF THE GENE IN IMMUNITY

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MS486 HIERARCHICAL ANALYSIS OF BIOMARKERS’ LEVELS, IN RELATION TO HYPERTENSION STATUS AMONG APPARENTLY HEALTHY PEOPLE V. Bountziouka, M. Bonou, A. Evangelopoulos, N. Vallianou, D. Politou, E. Bathrellou, A. Giotopoulou, D. Panagiotakos. Harokopio University, Athens, Greece Object: To evaluate the ability of various biomarkers in relation to hypertension among apparently healthy adults. Methods: 115 men (50±15 years) and 185 women (48±15 years) were randomly enrolled to the study. Discriminant analysis was used to assess the biomarkers that classify normotensive from treated or untreated hypertensive. Results: 74% (45±14 yrs) of the participants were normotensive, 14% (55±14 yrs) were untreated hypertensive and 12% (64±10 yrs) were hypertensive (i.e. SBP/DBP >140/90 mmHg or use of treatment). Untreated hypertension prevailed in 14% in both genders, while 15% of males and 10% of females were treated hypertensive. Education status was 14±4.7 yrs in normotensive, 10±5.5 yrs in treated and 11±5.7 yrs in untreated hypertensives (p < 0.001). Hierarchical analysis revealed that among the investigated biomarkers, gGt (lambda =0.949, p = 0.001), LDH (lambda =0.951, p = 0.001), uric acid (lambda =0.961, p = 0.004), cysteine (lambda=0.963, p = 0.005), ALP (lambda =0.974, p = 0.026), urea (lambda =0.974, p = 0.027) and C-RP (lambda =0.982, p = 0.078) were the factors that best classified participants in the aforementioned sub-groups. Fisher’s Discriminant equations revealed that cysteine, creatinine and C-RP were the factors that best characterized untreated hypertensives. The explained variability of the developed model was 30% (p < 0.001), while 63% was the correct classification rate. Conclusions: 50% of hypertensive individuals were untreated, with increased levels of biomarkers related to renal function. The latter, in addition to the high prevalence of hypertensives among low educated people, makes this group at extremely high-risk for developing atherosclerotic and other diseases. Health care practitioners should focus their interest in this group. MS487 CORRELATION OF IIB−IIIA INHIBITORS’ ADMINISTRATION AND PROGNOSIS OF PATIENTS WITH ACUTE MYOCARDIAL INFARCTION. AN AMI REGISTRY A. Gotsis, I.-A. Kerasidou, A. Gagalis, A. Risggits, D. Theodoridis, A. Koutsogianni, L. Borghi, P. Bozia, M. Benechoutsou, A. Panagiotidou. Cardiology, General Hospital of Komotini, Komotin´ı, Greece Objectives: The aim of this registry was to study the epidemiological and clinical characteristics of AMI (according AHA-WHF-ESC statement), in order to both prevent AMI and treat patients effectively. The aim of this study was to correlate GP IIb−IIIa inhibitors’ administration and prognosis of these patients. Methods: We studied 724 patients (506 men, aged 66.6 and 216 women, aged 73.3 years old) hospitalized in our department from January 2005 up to December 2007 with discharge diagnosis of AMI. A detailed medical history was taken emphasizing to CAD risk factors and previous cardiovascular events. Statistical analysis was performed by using SPSS 10.0. The methods used were x2 and Mann–Whitney test. Results: GP IIb−IIIa inhibitors were administrated in 41% of patients (29.5% of patients with STEMI and 52% of patients with NSTEMI). GP IIb−IIIa inhibitors’ administration was more frequent among younger patients, men with NSTEMI (p = 0.023), patients without previous stroke and non-hypertensive patients. There is not statistically significant difference in GP IIb−IIIa administration in diabetic and non-diabetic patients and those with or without previous AMI. Furthermore, statistical analysis proved no significant difference in frequency of minor hemorrhage events, after GP IIb−IIIa inhibitors’ administration. A detailed presentation of all above mentioned data appears in the table. Table: AMI and GP IIb−IIIa inhibitors’ administration p-value

Younger patients Without stroke history Without hypertension history Without diabetes history Without previous MI Mortality Minor hemorrhage events

AMI (total), n = 724

NSTEMI, n = 375

<0.001 0.004 0.044 0.152 0.816 <0.001 0.071

<0.001 <0.001 0.002 0.052 0.026 0.012 0.037

Conclusions: GP IIb−IIIa inhibitors were administrated in both high and medium risk patients without statistically significant difference in frequency of use. Patients who received GP IIb−IIIa inhibitors after AMI had better prognosis while no difference in minor hemorrhage events occurrence was observed.

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MS488 EFFECTS OF FLUVASTATIN ON TUMOR DEVELOPMENT AND PARAOXONASE-1 ACTIVITY IN RATS FED WITH CHOLESTEROL-RICH DIET J. Padra1 , P. Kertai2 , G. Foris ´ 1 , P. Ful ¨ op ¨ 1 , I. Seres1 , A. Olah ´ 3 , Z. Balogh1 , G. Paragh1 . 1 First Department of Medicine, Medical and Health Center, University of Debrecen, 2 Institute of Preventive Medicine, Medical and Health Science Center, University of Debrecen, 3 Department of Clinical Biochemistry and Molecular Pathology, Medical and Health Center, University of Debrecen, Debrecen, Hungary Recently human follow-up studies and animal experiments suggested a relationship between hypercholesterolemia (HC) and cancer. Moreover, statins may be beneficial for the prevention and treatment of cancer. Objectives: We aimed to establish an animal model, appropriate for the simultaneous study of HC and tumor development. Methods: The experiments were carried out on male and female FLF1 rats after 6 weeks of cholesterol-rich diet (CRD). Fluvastatin treatment and tumor inoculation were applied 21 days after the beginning of CRD. Tumor cells (He/De hepatocellular cell line) were inoculated under the renal capsule. At the 42th day blood samples were taken and tumor weights, serum lipids and PON1 activities were determined. Results: 1. CRD caused an elevation of LDL-C and HDL-C levels. 2. The PON1 activity was higher in male than in female rats, and the CRD decreased PON1 activity both in male and female rats, which was not altered by gonadectomy. 3. In female F1 hybrids the increase in serum LDL-C and HDL-C levels were more pronounced than in male rats. Gonadectomy had no effect on these sex-dependent differences. 4. CRD caused a significant increase in the development of the primary He/De tumor and its metastases. Tumor development had no effect on the elevated serum LDL-C and HDL-C levels, however, PON1 activity was significantly decreased. 5. Fluvastatin administration decreased both serum LDL-C and HDL-C levels and tumor development in the same rat. Conclusion: Our model is appropriate to study the effect of therapeutics against both hypercholesterolemia and cancer in the same rat. MS489 PLASMA FATTY ACID COMPOSITION IN PANCREATIC CARCINOMA M. Vecka, J. Macaˇ ´ sek, T. Krechler, M. Urbanek, ´ M. Zeman, A. Zˇ ak. ´ IVth Dept Int Med, 1st Medical Faculty, Charles University, Prague, Czech Republic Background: Pancreatic carcinoma (PC) is one of leading causes of gastrointestinal malignities in the Czech Republic. The nutrition factors playing a role in PC onset and development also include higher input of fat with lower intake of polyunsaturated fatty acids (PUFA) of n-3 family. Materials and Methods: The study included 62 patients (33M/29F) with pancreatic carcinoma and 39 control individuals (14M/25F). The fatty acid (FA) profile in plasma lipid classes was analysed by gas chromatography. Results: In plasma phospholipids, we found higher content of saturated and monounsaturated FA in the PC group, caused by higher contents of palmitic and oleic acids; changes in PUFA content included lower ratio of linoleic acid, which was reflected in decreased proportion of PUFA n-6 sum (all p < 0.0001) in PC group. The contents of eicosapentaenoic as well as a-linolenic acids were in PC group lower (both p < 0.0002). Lower content of linoleic acid was observed also in cholesteryl esters, whereas raised content of monounsaturated FA was found in both triacylglycerols and cholesteryl esters. Correlations of FA within the PC group revealed positive relationships between eicosapentaenoic acid and activity of cholinesterase, concentrations of total protein, albumin as well as with prealbumin (all p < 0.05). Conclusions: The changes in FA profile in patients with pancreatic cancer are probably of multifactorial origin. The factors involved are pathogenesis of tumor cachexia, enhanced turnover of free FA, inhibition of lipoprotein lipase and activation of de novo lipogenesis. Study was supported by the Research Project MSM 0021620820 MS490 LARGE-SCALE PHENOTYPING OF EMP3-KNOCKOUT MICE AT THE GERMAN MOUSE CLINIC REVEALS A SPECIFIC ROLE OF THE GENE IN IMMUNITY Z. Aherrahrou1 , T. Adler2,3 , V. Gailus-Durner2 , H. Fuchs2 , D.H. Busch3 , A. Wagner1 , M. Hrabe´ de Angelis2,4 , H. Schunkert1 , J. Erdmann1 . 1 Department 2 ¨ zu Lubeck, of Medicine II, Universitat ¨ Lubeck, ¨ German Mouse Clinic, 3 Institute of Experimental Genetics, Helmholtz Zentrum Munchen, ¨ Institute for 4 ¨ Munchen, Chair of Medical Microbiology, Technische Universitat ¨ Munchen, ¨ Experimental Genetics, Center of Life and Food Sciences Weihenstephan, ¨ Munchen, Technische Universitat ¨ Freising, Germany Introduction: Epithelial membrane protein 3 (EMP3) was previously reported to be involved in immunological reactions and in tumor as potential suppressor, as well as a candidate gene for cardiac dystrophic calcification (DCC). Given

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its assumed role in different disorders, we aimed to generate and perform a generalized phenotypic screening of EMP3-knockout mice. Methods: Knock-out targeting strategy was used to generate knockout mice for EMP3. Large-scale phenotyping screens were performed at the German Mouse Clinic to obtain a standardized and comprehensive way of phenotyping. The phenotype screens involved tests for nearly 320 parameters in different screening areas. For DCC, screening was performed in our laboratory using the freeze-thaw injury method. Results: EMP3-KO mice are viable and fertile. Under baseline conditions, an aberrant immunological phenotype was found with nearly 70% penetrance in mutant male mice. Specifically, a lower frequency of T cells and an inverse trend in B cells was observed. Also a higher frequency of B cells was seen in female mutant mice. For all the remaining screening, no genotype-specific differences were found. Also no calcification deposits were found in the EMP3-KO mice as response to injury. Conclusion: Large-scale phenotypic screening suggests a role of EMP3 under basal conditions in immunity. Differences in the proportion of various leukocyte subsets from the corresponding wild type were found. Further investigation is ongoing to demonstrate the role of EMP3 as tumor suppressor in a sensitized model. Finally, we excluded the EMP3 gene as candidate gene for DCC on the C57BL/6 genetic background. MS491 METABOLISM OF CHROMIUM COMPOUNDS IN RATS AND HUMANS P. Nielsen, K. Blaue, N. Laschinsky, G. Coxilha, N. Friedrich, K. Kottwitz. Department of Biochemistry and Molecular Biology II: Molecular Cell Biology, University Medical Center Hamburg, Hamburg, Germany Objectives: Chromium supplementation is discussed in the literature as a new way to prevent or treat insulin resistance and diabetes type 2. However, little is so far know on the biochemical mechanisms of Cr3+ action, and on the metabolismus of chromium from various pharmaceutical Cr compounds. Methods: Cr compounds used in food supplementation were labelled with 51 Cr and the absorption, retention and organ distribution was studied in rats after oral or parenteral administration. The absorption of Cr-picolinate and Cr-Dphenylalaninate was tested also in human volunteers using 51 Cr-whole-bodycounting. Results: Independent from the dose, the 7-d-whole body retention of 51 Cr after oral application was rather low (< 1%) from all compounds tested. Cr-picolinate, the market leader for Cr supplementation, is absorbed significantly better in rats and in also in humans, however, most of its Cr is directly lost into the urine indicating a different absorption mechanism from this stable Cr-complex. Cell culture experiments indicate a passive uptake of Cr in macrophages and hepatocytes. The 51 Cr whole-body-retention data in rats can be described in a 3-compartment model with typical half-lives of 0.2, 5, 100 days. The longer half-lives were similar for all Cr-compounds indicating the storage of comparable amounts of Cr in liver, muscle, fat, and bone. Conclusions: This study shows the limited bioavailability of oral chromium in rats and humans, especially of Cr-picolinate. This stable complex is absorbed as intact molecule to a higher degree compared to more ionic compounds, however, the retention is lower due to higher urine excretion. MS492 DOES RATIO OF WIDTH TO LENGTH OF LEFT ATRIAL APPENDAGE DETERMINE THE OCCURRENCE OF CEREBROVASCULAR STROKE EVENT? M. Jyotsna1 , S. Kaul2 , R. Narendra Kumar1 , G. Indrani1 . 1 Cardiology, 2 Neurology, Nizam’s Institute of Medical Sciences, Hyderabad, India Objective: To see the Left atrial appendage size (width and length) in ischemic cerebrovascular accident (CVA) patients. Design and Method: Transesophageal echocardiography (TEE) was done in ischemic CVA patients after initial stabilization. We compared width length ratio (WLR) calculated from the average of width and length of LAA observed in earlier study by John P et al as controls. We chosen this autopsy study as controls, as it is not possible to have TEE data in normal subjects. Results: Study group includes 143 patients (F:M::37:106) and control group was 400 subjects (F:M::200:200) from the previous study. The mean±std of width and length of LAA in females and males in study group were 1.99±0.67, 2.397±0.58 and 2.19±0.73, 2.49±0.66 respectively. In females, WLR is more (0.83±0.37) in study group than in control group (0.66±0.26) which is statistically significant (z = 4.06, P < 0.000001). So also for males (study group:control group: 0.88±0.39:0.71±0.28) which is also statistically significant (z = 7.03, P < 0.000001). So, broad and short LAA could determine the formation and dislodgement of thrombus resulting in embolic stroke. Conclusion: Higher the WLR of LAA, greater chances of embolic stroke. Reference(s) [1] John P. Veinot, Phillip J. Harrity, Federico Gentile, Bijoy K. Khandheria et al Anatomy of the Normal Left Atrial Appendage. A Quantitative Study of Age-Related Changes in 500 Autopsy Hearts: Implications for Echocardiographic Examination Circulation. 1997;96:3112–3115.

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MS493 EFFECT OF LOVASTATIN THERAPY AND WITHDRAWAL ON SERUM URIC ACID LEVEL IN TYPE II DIABETIC NEPHROPATHY N. Nezami1 , J. Safa2 , H. Argani1 , A. Ghorbanihaghjo3 , A.M. Vatankhah1 . 1 Drug Applied Research Center, 2 Department of Nephrology, 3 Biotechnology Research Center, Tabriz University (Medical Sciences), Tabriz, Iran Background: Recent studies have demonstrated that high uric acid level is a major risk factor of nephropathy and cardiovascular events in the patients with type II diabetes. We aimed to evaluate the serum uric acid (UA) level changes following treatment and withdrawal of lovastatin in patients with type II diabetic nephropathy (DN II). Methods: Thirty male patients with DN II were enrolled in the study. lovastatin, 20 mg/d, was administered for 90 days. Afterwards, lovastatin was withdrawn for the next 30 days. Blood samples were obtained at baseline, after 90 days of the intervention, and 30 days after withdrawal of lovastatin. Serum level of UA was determined by uricase/PAP method (an enzymatic color test). Lipid profile was assessed using commercial reagents. Results: After 90 days lovastatin intervention, Cholesterol (TC) and low-density lipoprotein (LDL-C) levels significantly decreased and high-density lipoprotein (HDL-C) level increased significantly (p < 0.001, 0.010 & 0.008), despite the unchanged level of triglyceride (TG). After 30 days of withdrawal, TC, TG and LDL-C levels were significantly increased (p < 0.001, 0.010, <0.001), but HDL-C level was not changed. Serum levels of UA were not changed during intervention and withdrawal of lovastatin. Conclusions: Lovastatin have not any effect on serum UA level in patients with DN II, and there is not any correlation between anti-lipidemic effect of lovastatin and its effect on serum UA. MS494 ROSUVASTATIN MODULATES GLUCOSE AND FFA-INDUCED PRO-FIBROTIC PATTERNS IN HUMAN MESANGIAL CELLS A. Solini, E. Santini, C. Rossi, A. Salvati, E. Ferrannini. Department of Internal Medicine, University of Pisa, Pisa, Italy Introduction: Persistent inflammation contributes to the progression of mesangial expansion toward fibrosis. Statins show anti-inflammatory effects in patients with chronic kidney disease. Methods: We explored the effect of rosuvastatin (RSV) on inflammatory and pro-fibrotic responses due to exposure to normal (NG) or high glucose (HG) or free fatty acid (FFA) of human mesangial cells (HMC) treated with Angiotensin-II (Ang-II). We measured MMP-2, MMP-9, TIMP-1, TIMP-2 expression (by realtime-PCR, reported as targeted/reference ratio) and activity (by zymography, expressed as AU), and TGFb, fibronectin and collagen-IV release by ELISA. Results: At NG, Ang-II dose-dependently downregulated MMP-2 (0.67±0.19 and 0.40±0.10 at 0.5 and 1mM, p < 0.01 vs basal); RSV partially reversed this effect (1.67±0.12 and 2.5±0.21 with RSV 1 and 5mM, both p < 0.005 vs Ang-II 1mM). Conversely, TIMP-2 (main MMP-2 inhibitor) and MMP-9 were upregulated by Ang-II (TIMP-2:1.17±0.14 and 1.87±0.18, p < 0.008 vs basal; MMP-9: 2.53±0.50 and 3.33±0.74, p < 0.05 vs basal) and down-regulated by RSV (TIMP-2: 1.03±0.09 and 0.70±0.15, p < 0.05 vs Ang-II 1mM; MMP-9: 3.37±0.14 and 2.20±0.10, p < 0.0005 vs basal); the effect of Ang-II on TIMP-1 was negligible. Some responses were potentiated by HG and FFA, being RSV able to partially revert them. MMP-2 and MMP-9 activities were coherent with their expression. RSV 1 and 5mM upregulated MMP-2 activity (NG: +20±6% and +37±9%), with a less defined effect on MMP-9. Following this MMP– TIMP modulation, TGFb increased after Ang-II (NG: from 217±19 to 337±24 and 391±21) and decreased after RSV (237±22 and 203±24 pg/ml/mg protein). Fibronectin and collagen-IV release showed a similar pattern in all the experimental conditions. Conclusions: Ang-II induces pro-fibrotic responses in HMC, mainly via a dysregulation of the MMP-2/TIMP-2 pattern. These effects, amplified by HG and FFA, were partially reverted by RSV, suggesting a potential new therapeutic application of this HMGCoA-reductase-inhibitor. MS495 PRETREATMENT EFFECT OF Cornus mas L. ON ALLOXAN-INDUCED DIABETES IN RATS S. Asgary1 , F. Shamsi1 , S. Kazemi1 , A. Adelneia1 , M. Rafieian2 . 1 Cardiovascular Research Center/Isfahan University of Medical Sciences, Isfahan, 2 Medicinal Plant Reserch Center Shahrekord, Shahrekord, Iran Background: Diabetes mellitus is a chronic and major endocrine disorder. phytochemicals present in medicinal plant may play a preventive role in the development of diabetes and diabetes-related complication. Cornus mas is one of those promising plants. This study was designed to investigate the pretreatment effects of fruit of cornus in diabetic rats. Methods: Diabetes mellitus was induced in 24 out of 32 male Wistar rats, using intraperitoneal (IP) injection of 120 mg/kg BW Alloxan. The diabetic rats were divided into three groups; (I) diabetic rats, (II) diabetic rats treated with cornus (2 g/daily), and (III) diabetic rats treated with Glibenclamide (0.6 mg/kg BW, daily). The remaining non-diabetic rats received neither alloxan nor the mentioned plants. 28 days before of induction of diabetes, total of groups were treated with ordinary diet except the second group, that received either diet