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Hypertriglyceridemia-induced acute pancreatitis in third trimester of pregnancy: A case report
Abstracts / Atherosclerosis 263 (2017) e111ee282
1924/2006 IN THE FRAMEWORK OF PREVENTION OF DYSLIPIDAEMIA AND CARDIOVASCULAR DISEASES Daniela Martini1, Beatrice Biasini1, Alessandra Dei Cas2, 3, Giorgio Bedogni4, Stefano Rossi1, Carlo Pruneti5, Giovanni Passeri6, Marilena Musci7, Ivana Zavaroni2, 3, Marco Ventura8, Marco Vitale9, Riccardo C. Bonadonna2, 3, Daniele Del Rio1. 1 The Laboratory of Phytochemicals in Physiology, Department of Food Science, University of Parma, Parma, Italy; 2 Department of Clinical and Experimental Medicine, Division of Endocrinology, University of Parma, Parma, Italy; 3 Azienda Ospedaliera Universitaria of Parma, Parma, Italy; 4 Clinical Epidemiology Unit, Liver Research Center, Basovizza, Trieste, Italy; 5 Department of Clinical and Experimental Medicine, Clinical Psychology Unit, University of Parma, Medical School Building, Parma, Italy; 6 Department of Clinical and Experimental Medicine, Building Clinica Medica Generale, University of Parma, Parma, Italy; 7 Department of Food Science, University of Parma, Parma, Italy; 8 Department of Life Sciences, Laboratory of Probiogenomics, University of Parma, Parma, Italy; 9 Department of Biomedical, Biotechnological and Translational Sciences (SBiBiT), Sport and Exercise Medicine Center(SEM), Parma, Italy Aim: To date a high number of requests for authorisation of health claims pursuant to Article 13(5) and 14 of Regulation (EC) No 1924/2006 has received a negative opinion from the European Food Safety Authority (EFSA). One of the most critical limitations is the choice of inappropriate outcome variables (OVs) and of their methods of measurement (MMs) in the studies supporting the substantiation of health claims. The present work reports the results of a critical evaluation of OV and MM used for the substantiation of health claims in the framework of prevention of dyslipidaemia and cardiovascular diseases. Methods: OVs and MMs were extracted from EFSA’s Guidance documents and Scientific Opinions on claim applications. Databases of references were created on PubMed to allow a specific critical analysis of all OVs and MMs. Results: Each OV and related MM was critically classified in one of the following categories: (i) appropriate alone; (ii) appropriate only in combination with other OV/MM; (iii) not appropriate per se; (iv) not appropriate in relation to the specific claimed effect, (v) not appropriate but useful as supportive evidence. Conclusions: The information provided in this document could serve as basis for EFSA to develop further guidance to applicants in the preparation of applications for authorization of health claims proposed under Regulation 1924/2006 in the framework of prevention of dyslipidaemia and cardiovascular diseases. In addition, it could be useful for applicants during the selection/design of human intervention studies aimed to substantiate such health claims. This project has received financial support from EFSA, Grant GP/EFSA/NUTRI/2014/01.
PO302. CYSTATIN C AS PROGNOSTIC MARKER OF CORONARY ARTERY DISEASE S. Panakala1, K. Kalyan Kumar2, K. Rambabu1, S. Venkata Rao1, M. Vijaya Bhaskar3, P. Jagannadha Rao4. 1 Dept of Biochemistry, Mamata medical college, Khammam, India; 2 Dept of Biochemistry Varun Arjun Medical college, Lucknow, India; 3 Dept of Biochemistry Parul Medical College, Vadodara, India; 4 Dept of Biochemistry and Medical Genetics, St.Matthews University, Grand Cayman, Cayman Islands Aim: Introduction: Cystatin C is a 13 KD protein composed of 120 amino acid residues which produce in all nucleate cells of the body. Cystatin C was a recognized marker of renal dysfunction, is gaining importance in dysfunction of other organs as well. Preliminary studies indicated a role for cystatin C as a projecting marker in coronary artery disease (CAD). CAD is the primary cause of death and has emerged as major health burden worldwide. The prevalence is increasing as well and is affecting younger age group. By 2020, 60% of the world's heart disease is expected to arise in India. The fundamental cause of CAD is atherosclerosis. There is a proof that both elastolytic cysteine proteases (cathepsins) and their inhibitors, an important one being cystatin C are implicated in the pathogenesis of atherosclerosis. Objectives: Aim to assess the of serum cystatin C levels in
e203
CAD and its clinical spectrum. The spectrum of CAD is Stable angina (SA), Unstable angina (UA) and Myocardial infarction (MI). Methods: Study group comprised of 145 patients diagnosed as having CAD based on clinical and bio-chemical criteria. Control group included 66 age and sex matched subjects (non CAD cases) using the above mentioned criteria. Results: In this study significant enhance of mean serum cystatin C levels were observed in CAD cases when compared with controls. Highest mean cystatin C values were observed in MI than UA and SA.Conclusion: Increased cystatin C levels may reflect CAD associated with inflammation and atherosclerosis. Serum cystatin C plays a significant role in the progress of CAD and it might have a role as a prognostic marker of CAD. Poster session: lipid and lipoprotein metabolism PO303. TRIGLYCERIDE BLOOD LEVELS AND METALLOPROTEINASES ACTIVITY IN ACUTE MYOCARDIAL INFARCTION PATIENTS Dimitrios Papadopoulos1, Haralambos Grassos2, Elias Sanidas1, John Barbetseas1. 1 Laiko Hospital, Athens, Greece; 2 Kat Hospital, Athens, Greece Aim: Matrix Metalloproteinases MMPs and their inhibitors TIMPs are key enzymes for myocardial collagen degradation in patients (pts) with acute myocardial infarction (AMI). The aim of this study was to investigate the influence of triglyceride blood levels on collagenolytic activity in pts with AMI. Methods: We measured MMP-1, TIMP-1, MMP-1/TIMP-1, plasma levels in 24 pts mean age (58,46+-13,9) yrs with AMI and classified them in two groups according to serum level of triglyceride in TRGL >150 mg%(TRIGL1) and in TRGL <150mg%(TRIGL-2) Plasma samples were collected at the time of admission in the hospital (0 hours) as well as 3h, 6h, 9h, 12h, 18h, 24h, 36h, 48h, 3days, 4d, 5d, 7d, 15d, 30d, thereafter and measured them by relevant ELISA kits. The mean latency between the onset of AMI and the admission in the hospital was 2.0+1.0 hours. All patients had no previous history of any other disease. For statistical analysis ANOVA rp.m.a and unpaired t-test were used. Data are expressed as mean values +-SD in ng/ml. p<0,05 was considered statistically signficant. Results: The mean blood values of collagenolytic enzymes in TRGL -1 group compared to the TRGL -2 were increased average by 10% for MMP-1 and reduced by -20% and by -4% for TIMP-1 and MMP-1/TIMP-1 complex respectively. Statistical significance was found by ANOVA rp.m.a within subjects alone for MMP-1, for TIMP-1 and for MMP-1/TIMP-1 complex respectively with 0,012>p>0,003. Conclusions: Hypertriglyceremia in pts with AMI increase MMP-1 activation and decrease expression of TIMP-1 leading to an imbalance between them making worse the ischemic injury.
PO304. HYPERTRIGLYCERIDEMIA-INDUCED ACUTE PANCREATITIS IN THIRD TRIMESTER OF PREGNANCY: A CASE REPORT Ioanna Polypathelli, Christos Demosthenous, Maria Gavra, Eleni Matsaridou, Glykeria Tzatzagou. Papageorgiou General Hospital, Thessaloniki, Greece Aim: Hypertriglyceridemia-induced acute pancreatitis during pregnancy is a rare condition occurring in approximately 3 in 10000 pregnancies and carries a dismal prognosis. It is known to be associated with an increase in estrogen. The mainstay of treatment includes dietary restriction of fat and lipid-lowering medications. Plasmapheresis has also been described. Methods: A 38-year-old Caucasian woman, Gravida 2, Para 1, was presented at the emergency department at 30 weeks of gestation for acute abdominal pain. The patient had a history of type 2 diabetes mellitus and preeclampsia during her first pregnancy. As the patient showed lipemic in blood on emergency room, tests were performed after high-speed centrifuging.
e204
Abstracts / Atherosclerosis 263 (2017) e111ee282
Results: Laboratory work up revealed triglyceridemia at 14440 mg/dl, total cholesterol of 1600 mg/dl, serum amylase 540 U/l and urine amylase 7620 U/l. Resistant hypocalcemia and exaggerated thrombocytosis at 1010/ul were also observed. Thrombocytosis was attributed to the inflammation. Cesarean section was performed and a healthy male infant was born. Intraoperative findings included milky peritoneal fluid collection. Abdominal ultrasonography showed cholelithiasis, while acute pancreatitis was confirmed by computed tomography scan. The patient was managed with nil per os, LMWH, intravenous fluids and antibiotics. Treatment with fenofibrate, atorvastatin and omega-3 fatty acids was started. The patient was stabilized, whereas her triglyceride levels fell to 521 mg/dl within 9 days. Laparoscopic cholecystectomy was performed on a later stage. Conclusions: Overall, detection of hypertriglyceridemia and pancreatitis in pregnancy can be challenging, while timely diagnosis and management of both conditions can help improve maternal and fetal outcome.
PO305. SPHINGOMYELINASE-INDUCED LDL AGGREGATION IS BLOCKED BY APOLIPOPROTEIN J MIMETIC PEPTIDE D-[113e122]APOJ ~ ez Llanos1, 2, Jose Luis Andrea Rivas Urbina1, Anna Rull1, Jordi Ordon Sanchez Quesada1. 1 Cardiovascular Biochemistry Department - Research Institute of the Hospital Sant Pau (IIB Sant Pau), Barcelona, Spain; 2 noma Biochemistry and Molecular Biology Department - Universitat Auto de Barcelona, Cerdanyola, Spain Aim: The use of mimetic peptides as potential therapeutic agents for atherosclerosis has emerged in the last years as a promising tool. The observation that a 10-residue class G* peptide from apolipoprotein J, namely D-[113e122]apoJ, possesses anti-inflammatory and anti-atherogenic properties prompted us to determine its effect on the aggregation process and structural properties of the LDL particles modified by SMase, an early event in the development of atherosclerosis. Methods: LDL particles with and without [113e122]apoJ peptide were incubated at 37ºC with SMase and their aggregation was analyzed at various time points by size-exclusion chromatography (SEC), native gradient gel electrophoresis (GGE), absorbance at 405 nm and cholesterol precipitation. Circular dichroism was also measured to determine structural changes in apoB and SDS-PAGE was performed to assess apoB degradation. Results: At a molar ratio of [113e122]apoJ peptide to apoB-100 (1:1), [113e122]apoJ blocked SMase-induced LDL aggregation and a full protection against LDL aggregation was observed up to 3h of incubation. All methods showed that [113e122]apoJ retarded the progression of SMaseinduced LDL aggregation with prolonged incubation times. SDS-PAGE showed that [113e122]apoJ also inhibited apoB degradation. No effect of [113 122]apoJ on apoB structure measured by circular dichroism was observed. Conclusions: These results demonstrate that [113e122]apoJ peptide prevents SMaseinduced LDL aggregation at an equimolar ratio and this opens the possibility of the use of this peptide as a therapeutic tool in the future.
PO306. SIALIDASE INHIBITORS SUPPRESS ATHEROGENIC MODIFICATION OF LDL Alexander Orekhov1, Nikita Nikiforov2. 1 Institute of Generel Pathology and Pathophysiology, Moscow, Russia; 2 Institute for Atherosclerosis Research, Moscow, Russia Aim: The crucial processes leading to atherogenic multiple modification of LDL is enzymatic modification (desialylation) of lipoprotein particles. Trans-sialidase discovered in human blood is responsible for desialylation of LDL. In vitro treatment of native LDL with trans-sialidase causes its desialylation and the appearance of atherogenic properties that is the ability of LDL to cause accumulation of intracellular lipids. In this study, we investigated the effect of natural products on the activity of the transsialidase isolated from human blood plasma.
Methods: For determination of trans-sialidase activity 3H-labeled LDL covalently bound to CNBr-activated sepharose was used as sialic acid donor. The intracellular accumulation of cholesterol was induced by incubation of cells with atherogenic modified LDL isolated from the blood of patients with atherosclerosis. Results: We have found that extracts of royal kombu, bladder wrack, yarrow, St. John's wort, onion and honey reduce trans-sialidase activity in vitro. The greatest inhibitory effect (90%) was found for pollen and garlic powder. Oral administration of pollen or garlic powder caused approximately 2-fold reduction in blood trans-sialidase activity determined ex vivo. The reduction of blood trans-sialidase activity was closely correlated with the reduction of the ability of plasma to cause cholesterol accumulation in cultured cells isolated from unaffected human aortic intima. Conclusions: Thus, some natural products are able to inhibit trans-sialidase activity in vitro and ex vivo. The inhibition of trans-sialidase leads to reduction of atherogenic properties of blood plasma. These results could provide a basis for the development of new approaches to prevention and treatment of human atherosclerosis.
PO307. SEASONAL CHANGES OF SIALIDASE ACTIVITY Alexander Orekhov1, Nikita Nikiforov2. 1 Institute of Generel Pathology and Pathophysiology, Moscow, Russia; 2 Institute for Atherosclerosis Research, Moscow, Russia Aim: We have discovered multiple-modified low density lipoprotein (mLDL) in the blood of atherosclerotic patients. mLDL is able to induce accumulation of lipids in human monocytes and in the smooth muscle cells cultured from unaffected intima of human aorta. Along with small size, high density and increased electronegative charge, mLDL is characterized by decreased level of sialic acid. Negative correlation between the level of sialic acid in LDL and its ability to induce accumulation of intracellular cholesterol has been found. Thus, desialylation is an atherogenic modification of LDL. In the blood of atherosclerotic patients we have discovered a soluble enzyme belonging by its donor-acceptor properties to trans-sialidases. Methods: Lipoprotein-deficient serum obtained by ultracentrifugation was used for isolation of trans-sialidase by the affinity chromatography with a-2,8-Neu5Ac-sepharose. After washing, sorbent-bound protein fraction was eluted with 50 mM sialic acid. Results: We have revealed seasonal differences in trans-sialidases activity in blood serum of 61% observed patients. It is known that coronary heart disease risk increases in winter. More than 43% of patients had higher level of trans-sialidase activity in winter as compared with summer period. The opposite situation occurred in 17% of patients. Thereby we could see that the most part of observed people have higher blood serum atherogenicity in winter period. This correlates well with seasonal dynamics of heart disease risk. Conclusions: Thus, trans-sialidases activity and concentration of mLDL in human blood serum can serve as the additional factors for estimation of cardiovascular disease risk.
PO308. THE CHARACTERISTICS OF BLOOD LIPIDS OF MENOPAUSAL WOMEN WITH THE COMPENSATED HYPOTHYROIDISM (TSH ¼0,4 e 2,5 MU/L) WHICH RESULTS FROM AUTOIMMUNE THYROIDITIS Rymar, Svetlana Mustafina, Lilya Yulua Malyshenko, Oksana Shcherbakova. Regional Clinical Hospital of Kaliningrad region, Kaliningrad, Russia Aim: To study the lipid profile, including of non-high-density lipoproteins cholesterol (non-HDL-C), in postmenopausal women with compensated hypothyroidism (TSH ¼0,4 e 2,5 mU/L). Methods: The causes of hypothyroidism included Hashimoto thyroiditis (HT).109 women with HT participated in the study (mean ± sd, age (57.4 ± 7.7) years, disease duration (8.0 ± 6.4) years, the duration of
1924/2006 IN THE FRAMEWORK OF PREVENTION OF DYSLIPIDAEMIA AND CARDIOVASCULAR DISEASES Daniela Martini1, Beatrice Biasini1, Alessandra Dei Cas2, 3, Giorgio Bedogni4, Stefano Rossi1, Carlo Pruneti5, Giovanni Passeri6, Marilena Musci7, Ivana Zavaroni2, 3, Marco Ventura8, Marco Vitale9, Riccardo C. Bonadonna2, 3, Daniele Del Rio1. 1 The Laboratory of Phytochemicals in Physiology, Department of Food Science, University of Parma, Parma, Italy; 2 Department of Clinical and Experimental Medicine, Division of Endocrinology, University of Parma, Parma, Italy; 3 Azienda Ospedaliera Universitaria of Parma, Parma, Italy; 4 Clinical Epidemiology Unit, Liver Research Center, Basovizza, Trieste, Italy; 5 Department of Clinical and Experimental Medicine, Clinical Psychology Unit, University of Parma, Medical School Building, Parma, Italy; 6 Department of Clinical and Experimental Medicine, Building Clinica Medica Generale, University of Parma, Parma, Italy; 7 Department of Food Science, University of Parma, Parma, Italy; 8 Department of Life Sciences, Laboratory of Probiogenomics, University of Parma, Parma, Italy; 9 Department of Biomedical, Biotechnological and Translational Sciences (SBiBiT), Sport and Exercise Medicine Center(SEM), Parma, Italy Aim: To date a high number of requests for authorisation of health claims pursuant to Article 13(5) and 14 of Regulation (EC) No 1924/2006 has received a negative opinion from the European Food Safety Authority (EFSA). One of the most critical limitations is the choice of inappropriate outcome variables (OVs) and of their methods of measurement (MMs) in the studies supporting the substantiation of health claims. The present work reports the results of a critical evaluation of OV and MM used for the substantiation of health claims in the framework of prevention of dyslipidaemia and cardiovascular diseases. Methods: OVs and MMs were extracted from EFSA’s Guidance documents and Scientific Opinions on claim applications. Databases of references were created on PubMed to allow a specific critical analysis of all OVs and MMs. Results: Each OV and related MM was critically classified in one of the following categories: (i) appropriate alone; (ii) appropriate only in combination with other OV/MM; (iii) not appropriate per se; (iv) not appropriate in relation to the specific claimed effect, (v) not appropriate but useful as supportive evidence. Conclusions: The information provided in this document could serve as basis for EFSA to develop further guidance to applicants in the preparation of applications for authorization of health claims proposed under Regulation 1924/2006 in the framework of prevention of dyslipidaemia and cardiovascular diseases. In addition, it could be useful for applicants during the selection/design of human intervention studies aimed to substantiate such health claims. This project has received financial support from EFSA, Grant GP/EFSA/NUTRI/2014/01.
PO302. CYSTATIN C AS PROGNOSTIC MARKER OF CORONARY ARTERY DISEASE S. Panakala1, K. Kalyan Kumar2, K. Rambabu1, S. Venkata Rao1, M. Vijaya Bhaskar3, P. Jagannadha Rao4. 1 Dept of Biochemistry, Mamata medical college, Khammam, India; 2 Dept of Biochemistry Varun Arjun Medical college, Lucknow, India; 3 Dept of Biochemistry Parul Medical College, Vadodara, India; 4 Dept of Biochemistry and Medical Genetics, St.Matthews University, Grand Cayman, Cayman Islands Aim: Introduction: Cystatin C is a 13 KD protein composed of 120 amino acid residues which produce in all nucleate cells of the body. Cystatin C was a recognized marker of renal dysfunction, is gaining importance in dysfunction of other organs as well. Preliminary studies indicated a role for cystatin C as a projecting marker in coronary artery disease (CAD). CAD is the primary cause of death and has emerged as major health burden worldwide. The prevalence is increasing as well and is affecting younger age group. By 2020, 60% of the world's heart disease is expected to arise in India. The fundamental cause of CAD is atherosclerosis. There is a proof that both elastolytic cysteine proteases (cathepsins) and their inhibitors, an important one being cystatin C are implicated in the pathogenesis of atherosclerosis. Objectives: Aim to assess the of serum cystatin C levels in
e203
CAD and its clinical spectrum. The spectrum of CAD is Stable angina (SA), Unstable angina (UA) and Myocardial infarction (MI). Methods: Study group comprised of 145 patients diagnosed as having CAD based on clinical and bio-chemical criteria. Control group included 66 age and sex matched subjects (non CAD cases) using the above mentioned criteria. Results: In this study significant enhance of mean serum cystatin C levels were observed in CAD cases when compared with controls. Highest mean cystatin C values were observed in MI than UA and SA.Conclusion: Increased cystatin C levels may reflect CAD associated with inflammation and atherosclerosis. Serum cystatin C plays a significant role in the progress of CAD and it might have a role as a prognostic marker of CAD. Poster session: lipid and lipoprotein metabolism PO303. TRIGLYCERIDE BLOOD LEVELS AND METALLOPROTEINASES ACTIVITY IN ACUTE MYOCARDIAL INFARCTION PATIENTS Dimitrios Papadopoulos1, Haralambos Grassos2, Elias Sanidas1, John Barbetseas1. 1 Laiko Hospital, Athens, Greece; 2 Kat Hospital, Athens, Greece Aim: Matrix Metalloproteinases MMPs and their inhibitors TIMPs are key enzymes for myocardial collagen degradation in patients (pts) with acute myocardial infarction (AMI). The aim of this study was to investigate the influence of triglyceride blood levels on collagenolytic activity in pts with AMI. Methods: We measured MMP-1, TIMP-1, MMP-1/TIMP-1, plasma levels in 24 pts mean age (58,46+-13,9) yrs with AMI and classified them in two groups according to serum level of triglyceride in TRGL >150 mg%(TRIGL1) and in TRGL <150mg%(TRIGL-2) Plasma samples were collected at the time of admission in the hospital (0 hours) as well as 3h, 6h, 9h, 12h, 18h, 24h, 36h, 48h, 3days, 4d, 5d, 7d, 15d, 30d, thereafter and measured them by relevant ELISA kits. The mean latency between the onset of AMI and the admission in the hospital was 2.0+1.0 hours. All patients had no previous history of any other disease. For statistical analysis ANOVA rp.m.a and unpaired t-test were used. Data are expressed as mean values +-SD in ng/ml. p<0,05 was considered statistically signficant. Results: The mean blood values of collagenolytic enzymes in TRGL -1 group compared to the TRGL -2 were increased average by 10% for MMP-1 and reduced by -20% and by -4% for TIMP-1 and MMP-1/TIMP-1 complex respectively. Statistical significance was found by ANOVA rp.m.a within subjects alone for MMP-1, for TIMP-1 and for MMP-1/TIMP-1 complex respectively with 0,012>p>0,003. Conclusions: Hypertriglyceremia in pts with AMI increase MMP-1 activation and decrease expression of TIMP-1 leading to an imbalance between them making worse the ischemic injury.
PO304. HYPERTRIGLYCERIDEMIA-INDUCED ACUTE PANCREATITIS IN THIRD TRIMESTER OF PREGNANCY: A CASE REPORT Ioanna Polypathelli, Christos Demosthenous, Maria Gavra, Eleni Matsaridou, Glykeria Tzatzagou. Papageorgiou General Hospital, Thessaloniki, Greece Aim: Hypertriglyceridemia-induced acute pancreatitis during pregnancy is a rare condition occurring in approximately 3 in 10000 pregnancies and carries a dismal prognosis. It is known to be associated with an increase in estrogen. The mainstay of treatment includes dietary restriction of fat and lipid-lowering medications. Plasmapheresis has also been described. Methods: A 38-year-old Caucasian woman, Gravida 2, Para 1, was presented at the emergency department at 30 weeks of gestation for acute abdominal pain. The patient had a history of type 2 diabetes mellitus and preeclampsia during her first pregnancy. As the patient showed lipemic in blood on emergency room, tests were performed after high-speed centrifuging.
e204
Abstracts / Atherosclerosis 263 (2017) e111ee282
Results: Laboratory work up revealed triglyceridemia at 14440 mg/dl, total cholesterol of 1600 mg/dl, serum amylase 540 U/l and urine amylase 7620 U/l. Resistant hypocalcemia and exaggerated thrombocytosis at 1010/ul were also observed. Thrombocytosis was attributed to the inflammation. Cesarean section was performed and a healthy male infant was born. Intraoperative findings included milky peritoneal fluid collection. Abdominal ultrasonography showed cholelithiasis, while acute pancreatitis was confirmed by computed tomography scan. The patient was managed with nil per os, LMWH, intravenous fluids and antibiotics. Treatment with fenofibrate, atorvastatin and omega-3 fatty acids was started. The patient was stabilized, whereas her triglyceride levels fell to 521 mg/dl within 9 days. Laparoscopic cholecystectomy was performed on a later stage. Conclusions: Overall, detection of hypertriglyceridemia and pancreatitis in pregnancy can be challenging, while timely diagnosis and management of both conditions can help improve maternal and fetal outcome.
PO305. SPHINGOMYELINASE-INDUCED LDL AGGREGATION IS BLOCKED BY APOLIPOPROTEIN J MIMETIC PEPTIDE D-[113e122]APOJ ~ ez Llanos1, 2, Jose Luis Andrea Rivas Urbina1, Anna Rull1, Jordi Ordon Sanchez Quesada1. 1 Cardiovascular Biochemistry Department - Research Institute of the Hospital Sant Pau (IIB Sant Pau), Barcelona, Spain; 2 noma Biochemistry and Molecular Biology Department - Universitat Auto de Barcelona, Cerdanyola, Spain Aim: The use of mimetic peptides as potential therapeutic agents for atherosclerosis has emerged in the last years as a promising tool. The observation that a 10-residue class G* peptide from apolipoprotein J, namely D-[113e122]apoJ, possesses anti-inflammatory and anti-atherogenic properties prompted us to determine its effect on the aggregation process and structural properties of the LDL particles modified by SMase, an early event in the development of atherosclerosis. Methods: LDL particles with and without [113e122]apoJ peptide were incubated at 37ºC with SMase and their aggregation was analyzed at various time points by size-exclusion chromatography (SEC), native gradient gel electrophoresis (GGE), absorbance at 405 nm and cholesterol precipitation. Circular dichroism was also measured to determine structural changes in apoB and SDS-PAGE was performed to assess apoB degradation. Results: At a molar ratio of [113e122]apoJ peptide to apoB-100 (1:1), [113e122]apoJ blocked SMase-induced LDL aggregation and a full protection against LDL aggregation was observed up to 3h of incubation. All methods showed that [113e122]apoJ retarded the progression of SMaseinduced LDL aggregation with prolonged incubation times. SDS-PAGE showed that [113e122]apoJ also inhibited apoB degradation. No effect of [113 122]apoJ on apoB structure measured by circular dichroism was observed. Conclusions: These results demonstrate that [113e122]apoJ peptide prevents SMaseinduced LDL aggregation at an equimolar ratio and this opens the possibility of the use of this peptide as a therapeutic tool in the future.
PO306. SIALIDASE INHIBITORS SUPPRESS ATHEROGENIC MODIFICATION OF LDL Alexander Orekhov1, Nikita Nikiforov2. 1 Institute of Generel Pathology and Pathophysiology, Moscow, Russia; 2 Institute for Atherosclerosis Research, Moscow, Russia Aim: The crucial processes leading to atherogenic multiple modification of LDL is enzymatic modification (desialylation) of lipoprotein particles. Trans-sialidase discovered in human blood is responsible for desialylation of LDL. In vitro treatment of native LDL with trans-sialidase causes its desialylation and the appearance of atherogenic properties that is the ability of LDL to cause accumulation of intracellular lipids. In this study, we investigated the effect of natural products on the activity of the transsialidase isolated from human blood plasma.
Methods: For determination of trans-sialidase activity 3H-labeled LDL covalently bound to CNBr-activated sepharose was used as sialic acid donor. The intracellular accumulation of cholesterol was induced by incubation of cells with atherogenic modified LDL isolated from the blood of patients with atherosclerosis. Results: We have found that extracts of royal kombu, bladder wrack, yarrow, St. John's wort, onion and honey reduce trans-sialidase activity in vitro. The greatest inhibitory effect (90%) was found for pollen and garlic powder. Oral administration of pollen or garlic powder caused approximately 2-fold reduction in blood trans-sialidase activity determined ex vivo. The reduction of blood trans-sialidase activity was closely correlated with the reduction of the ability of plasma to cause cholesterol accumulation in cultured cells isolated from unaffected human aortic intima. Conclusions: Thus, some natural products are able to inhibit trans-sialidase activity in vitro and ex vivo. The inhibition of trans-sialidase leads to reduction of atherogenic properties of blood plasma. These results could provide a basis for the development of new approaches to prevention and treatment of human atherosclerosis.
PO307. SEASONAL CHANGES OF SIALIDASE ACTIVITY Alexander Orekhov1, Nikita Nikiforov2. 1 Institute of Generel Pathology and Pathophysiology, Moscow, Russia; 2 Institute for Atherosclerosis Research, Moscow, Russia Aim: We have discovered multiple-modified low density lipoprotein (mLDL) in the blood of atherosclerotic patients. mLDL is able to induce accumulation of lipids in human monocytes and in the smooth muscle cells cultured from unaffected intima of human aorta. Along with small size, high density and increased electronegative charge, mLDL is characterized by decreased level of sialic acid. Negative correlation between the level of sialic acid in LDL and its ability to induce accumulation of intracellular cholesterol has been found. Thus, desialylation is an atherogenic modification of LDL. In the blood of atherosclerotic patients we have discovered a soluble enzyme belonging by its donor-acceptor properties to trans-sialidases. Methods: Lipoprotein-deficient serum obtained by ultracentrifugation was used for isolation of trans-sialidase by the affinity chromatography with a-2,8-Neu5Ac-sepharose. After washing, sorbent-bound protein fraction was eluted with 50 mM sialic acid. Results: We have revealed seasonal differences in trans-sialidases activity in blood serum of 61% observed patients. It is known that coronary heart disease risk increases in winter. More than 43% of patients had higher level of trans-sialidase activity in winter as compared with summer period. The opposite situation occurred in 17% of patients. Thereby we could see that the most part of observed people have higher blood serum atherogenicity in winter period. This correlates well with seasonal dynamics of heart disease risk. Conclusions: Thus, trans-sialidases activity and concentration of mLDL in human blood serum can serve as the additional factors for estimation of cardiovascular disease risk.
PO308. THE CHARACTERISTICS OF BLOOD LIPIDS OF MENOPAUSAL WOMEN WITH THE COMPENSATED HYPOTHYROIDISM (TSH ¼0,4 e 2,5 MU/L) WHICH RESULTS FROM AUTOIMMUNE THYROIDITIS Rymar, Svetlana Mustafina, Lilya Yulua Malyshenko, Oksana Shcherbakova. Regional Clinical Hospital of Kaliningrad region, Kaliningrad, Russia Aim: To study the lipid profile, including of non-high-density lipoproteins cholesterol (non-HDL-C), in postmenopausal women with compensated hypothyroidism (TSH ¼0,4 e 2,5 mU/L). Methods: The causes of hypothyroidism included Hashimoto thyroiditis (HT).109 women with HT participated in the study (mean ± sd, age (57.4 ± 7.7) years, disease duration (8.0 ± 6.4) years, the duration of