172 Post-congress. Kos

Poster session

30% TO 40% FAT IN THE DIET OF NORMOLIPIDEMIC MEN BY SUPPLEMENTATION WITH MONO AND/OR POLYUNSATURATED OILS: WHERE ARE THE LIMITS FOR BENEFICIAL EFFECTS ON LIPEMIA?

B. Delplanque, B. Le Roy, I. Jusselin, A. Ruelland, C. Morta. Laboratoire de Physiologie de la Nutrition. Universit~ Paris Sud 91405 ORSAY; Facult~ des Sciences Pharmaceutiques. Uniuersit~ de Rennes I 35000 Rennes, France

Four unsaturated diets at 2 levels o f fat [30% and 40%] were studied in 29 control mazes (BMi: 22+3, TC: 190+28 mg/dl, TG: 80+30 mg/dl, HDLC: 44+30 mg/dl) after specific modification of the visible fat consumption by using different vegetable oils in the diet: -two Oleic acid rich diets, by using oils containing more than 70%) MUFA: OL-I 74% and OL-I! 80%, but differing by the proportions of PUFA: 17% and 6% respectively. -one Linoleic acid rich diet, using Sunflower Oil (Sun: 22% MUFA, 65% PUFA) and -one Oleic/Linoleic intermediate diet, using a blend of oils (Isio: 41% MUFA, 47% PUFA).

The 30% fat basal diet was characterized by a low caloric intake (2022 +210 Kcal), and a consumption of 66 g fat/day, including 30 g of the specific vegetable oil studied. Supplementation with the corresponding oil to obtain a 40% fat in the diet (109 g/day), induced an increase of the caloric intake to 2411+410 Kcal, an increase of both Unsaturated (70 g vs 40 g/day) and Saturated FA (39 g os 26 g/day), and finally resulted in a 20% increase of the Unsaturated (MUFA + PUFA)/Saturated ratio ( 1.8 os 1.5). Subjects were assignated to six weeks study periods for each fat level and oil.

Results indicate that in these normal subjects, an increased consumption of unsaturated vegetable oils to reach 40°/, of fat in the diet, did not increase plasma cholesterol, nor triglycerides, but induced an increase of HDLC (HDL3), and a decrease of LDLC.

Considering the intake of MUFA and PUFA expressed in grammes per day, independently of the % (30 or 40) of total fat consumption, we verified the strong negative correlation between PUFA and LDLC (r = -0.82, p < 0.00I) and the positive one between MUFA and HDLC (r = +0.80 p < 0.001 ). Considering the specific effect of each type of oil in these diets -1) The most favorable effects on plasma lipids were observed with 30% and 40% lsio diet, which showed TC and LDLC levels as low as those seen with the high PUFA diet, but kept HDLC levels as high as those seen with the high MUFA diets, inducing the best HDLC/LDLC ratio. -2) The high MUFA OL-I diet induced lower levels of TC and LDLC than the other high MUFA diet OL-II, and this can be related to the higher level of PUFA provided by OL-L -3) The minimum daily MUFA intake level needed to obtain a beneficial effect on HDLC is reached with diets providing 28 g of MUFA, whatever could be the levels of PUFA (even with high PUFA Sun 40%) fat diet) -4) The minimum daily PUFA intake level needed to obtain a beneficial effect on LDLC is reached with diets providing 13.5 g of PUFA, even in the presence of high levels of MUFA (OL-I 40% fat diet) -5) Then, the most beneficial effects on lipemia obtained with the lsio diets could be explained partly, by a daily intake higher than 28 g of MUFA and 14g of PUFA, inducing the best Anti-Atherogenic plasma HDLC/LDLC Index, at any level of total fat consumption: 30% or 40%. This work was supported by grants from LESIEUR

P o s t e r s e s s i o n

BIOLOGICALLY ACTIVE LIPIDS FROM OLIVE OIL WITH ANTIATHEROGENETIC ACTION

C.A. Demopoulos 1 , S. Antonopoulou 2, T.N. Nomikos 1 . 1Faculty o f Chemistry, National & Kapodistrian Unioersity o f Athens, Athens; 2Department o f Dietetics, Harokopio University o f Home Economics, Athens. Greece

In recent studies we have shown that Platelet-Activating Factor (PAF) is implicated in atheromatosis generation and its inhibitors display protective action against atherogenesis. The objective of this work was to investigate the existence of biologically active lipids in olive oil in comparison with seed oils in order to explain the protective role of olive oil against cardiovascular diseases. Olive oil samples were from the same Greek area but extracted with different ways. The seed oils tested were soybean, corn, sunflower, cottonseed and sesame oils purchased from Greek market. Total lipids were extracted from oils with organic solvents, separated into classes and further more in species by HPLC. The HPLC columns used were normal and reverse phase as well as cation exchange with the appropriate solvent systems. Each lipid fraction from HPLC separation along with total, polar and neutral lipid samples from oils were tested in vitro for their ability

to induce washed rabbit platelet aggregation and/or to inhibit PAF and thrombin induced aggregation in aggregometer. Additionally, desensitization and cross-desensitization experiments along with experiments with specific inhibitors of each aggregation way -namely, BN 52021, indomethacine and the enzymic system ofcreatine phosphate/kinase of crcatine phosphate-were also used in order to specify the way by which biologically active lipids induce aggregation. In the olive oils, we detected inhibitors as well as lipid fractions which induced aggregation in neutral, glyco- and phospholipids but none of them was acting through PAF way only. The most potent agonist was found in phospholipids having similar chromatographic behavior with the one of PAF and its structure was further studied with electrospray mass spectrometry. This agonist was not found in the seed oils tested with the exception of soybean oil (about 20 times less active) and sesame oil (about 10 times less active). Comparison of the IC50 values (concentration which induce 50% inhibition against PAF I × 10 -l° M, final concentration) expressed in Itl o f initial sample showed that IC5o values for total and polar lipids of olive oils, 0.18 and 25 p.l, respectively, were lower than the ones of seed oils, 0.27 and 137.5 Ittl, respectively. The IC50 values for neutral lipids were approximately the same for olive and seed oils. The above results shows that olive oils in comparison to seed oils contain lipids with more potent activity to inhibit PAF action, as inhibitors or weak agonists. These findings support the hypothesis that the existence of these lipid compounds plays important role in the protective effect of olive oil in atheromatosis generation.

This work was supported by the General Secretiate for Research and Technology of the Ministry of Development.

TYROSOL, AN OLIVE OIL BIOPHENOL, PROTECTS INTESTINAL CULTURED CELLS CACO-2 AGAINST OXIDIZED-LOW DENSITY LIPOPROTEIN-INDUCED INJURY

C. Giovannini 1, E. Straface 3, E. Coni 2, M. De Vincenzi 2, R. Masella 1 . /Metabolism and Pathological Biochemistry. Lab.: "Food Lab.: 3 Ultrastructures Lab., Istituto Superiore di SanitY-Rome. ltaly

Low-density lipoproteins, undergoing oxidative modifications (ox-LDL), lead to the production of cytotoxic reactive species, e.g. lipoperoxides, caus- ing tissue injury and subsequent various pathologies including atherosclero- sis and intestinal diseases. The antioxidant intake with the diet presumably plays a relevant role in the prevention of oxidative damage. In particular. phenolic antioxidants, present in virgin olive oil, can protect against oxidative stress. Thus, in order to analyze the oxidative damage induced by ox- LDL to intestinal mucosa, we evaluated morphological and functional changes induced in the human colon adenocarcinoma cell line Caco- 2. Cytotoxicity was evaluated considering proliferation cell activity ([4C thymidine incorporation), cell membrane integrity (lactate dehydrogenase release), cell morphological and ultrastructural alterations (scanning electron microscopy), cytoskeleton organizzation (fluorescence microscopy obser- vation) and evaluation of apoptotic cells (Hoechst staining). Furthermore, we examined the protective effects exerted on these cells by tyrosol, (p - hydroxyphenylethanol), the major phenolic compound present in olive oil. Caco-2 cell treatment with ox-LDL resulted in cytostatic and cytotoxic effects characterized by a series of morphological and functional alterations: membrane damage, modifications of cytoskeleton network, microtubular disorganization, loss of cell-cell and cell-substrate contacts, cell detachment and cell death. The ox-LDL-induced alterations were almost completely prevented by preincubating Caco-2 cells with tyrosol. The results obtained. provide evidence that physiologic oxidative-stress inducers such as ox-LDL exert a cytotoxic effect on an in vitro enterocyte-like cell system, and that phenolic compounds with antioxidant properties, such as those contained in virgin olive oil, may exert a protective effect. Our results confirm the relevance of dietary intake of olive oil biophenols based on their capability to lower the risk of reactive oxygen metabolite-mediated diseases such as atheroselerosis and inflammatory bowel disease.

PALEONUTRITION AND MEDITERRANEAN DIET ORIGIN

A. Corominas. Hospital Unioersitari Germans Trias i Pujol, L C.S. Departament Biologia CeLlular i Fisiologia Unioersitat Autfnoma de Barcelona, Spain

Introduction: To implant a reflection over the origin of the Mediterranean diet base on the evidences of the archaeological findings in a Mediter- ranean site in Catalona called Seriny/t (Girona), where since 1924 a lot of material has been collected. The material has been widely published by

71st EAS Congress and Satellite Symposia

Post-congress, Kos Poster session 173

J.M. Corominas who studied the problem through the descriptive, cultural, paleoanthropological and partially paleopathologic point of view.

Material: Moreover the temporary dating realized with CI4, it is im- portant to consider the concept of paleonutrition, lifestyle of the habitants (Homo Sapiens Sapiens) from the excavations realized and attempt to establish the possible onset of the Mediterranean diet. We took nutritional reference points on the aboriginal people, and have been considered the studies performed by STINGER over the genetic basis protein polimor- phisms, Mytocondrial and nuclear DNA. EATON in 1996 reported that our genetic inheritance (99%) proceed from 2 million years, and that a 99.9% was constituted before the development of agriculture, about 10.000 years ago. So that nowadays our genes are similar to those from the Neolitic habitants of the Mediterranean see with a different lifestyle, who did not complaint about the so called Civilization Diseases such as acute myocardial infarct, atherosclerosis, pancreatitis hypertension, diabetes mellitus !I, and some type of cancer. We could say that we have genes from yesterday and lifestyle from today (hunter-gatherer lifestyle vs sedentary consumer- civility), in the area of Catalonia exists rests in Puig d'en Roca, Tautaou Coon de Mollet, Reclau-oiver. the mandible of Banyoles. Since this data, started in the whole Iberian Peninsula and the rest of the Mediterranean see the practice of the agriculture, cattle raising. It is essential for the onset o f the Mediterranean diet the appearance of ceramics, that represents the onset of the culinary practice. Palatal and sybaritic gastronomy are considered to be here introduced for the first time and we could also consider the appearance of the chronobiological phenomena and circadian rhythm of meals.

Conclusions: We could consider the following requirements to accept the existence of the Mediterranean Diet: I) Established circadian rhythm, 2) 75% of vegetables in meals 3) Cereals with bread grains partially 4) Use of Olive oil, 5) Vine consumption

A table: with the stages of the Mediterranean Diet is included.

References

It] Corominas JM, Corominas A. La alimentacitn del hombre prehisttrico. Unpub- lished.

[2] Eaton S, Konner M. Paleolithic nutrition. Ntnv Engl. Jour. Med, 312: 283-89. Jan 1983

THE STRESS TEST BASED OBJECTIVIZATION OF THE MYOCARDIAL ISCHEMIA OF DIFFERENT GENESIS

V.K. Tashchuc, S.I. Grechko, L.P. Sydorchuk, O.Yu. Polishchuk. Bucovinian State Medical Academy, Cherniotci, Ukraine

60 patients of 24 to 67 years old (mean age - 44.6 + 1.9) complaining of thoracic pain with changes in ending part o f ventricular complex or rhythm disorders on ECG were observed. 22 of them had stable angina (SA), 19 patients with unstable angina (UA) 23 patients with non-Q-wave myocardial infarction (MI), All patients underwent transoesophageal cardiostimulation (TECS) according to traditional protocol. The stimulation halting frequency and quantity figures of myocardial ischemia were analyzed: total ST segment depression (EST), number of ischemically changed leads (NST) and the average depth of ST segment depression (AST). The average stimulation halting frequency index in none-Q-wave MI patients was 133.9 + 3.7 imp/min, in UA patients - 138.0 + 6.4 imp/min (p > 0.5) and SA patients - 156.3 + 2.5 imp/rain respectively (p < 0.001). EST in female patients with UA was lower (4.65 + 0.21 mm) in comparison with the figures of myocardial infarction patients - 5.76 + 0.8 mm (p < 0.001). The same difference in male was between the MI - 4.78 + 0.61 mm and UA - 5.24 + 0.12 (p > 0.05) and significant difference in cases o f S A - 2.78 + 0.47 mm (p > 0.05). Similar tendencies were observed in male patients with higher figures. The most significant decrease of EST within SA - 2.39 + 0.24 mm, less significant in UA patients - 5.45 + 0.35 mm (p < 0.001) and M1 - 6.08 + 0.31 mm (p > 0.001). Average ST segment dislocation depth exceeded in female patients with MI - 1.26 + 0.08 mm and UA - 1.23 + 0.98 (p > 0.5), lower figures were observed in SA patients - 1.24 + 0.05 mm (p > 0.5). In male patients the corresponding figures were SA - 1.00 + 0.09, Mi - 1.33 + 0.05 (p > 0.001) and UA - 1.37 + 0.07 (p > 0.001). Differences in the stress-test results in M! and UA patients and functional disorders may be used for differential diagnosis.

HYPOLIPIDEMIC REMEDY VITOL IN CORONARY HEART DISEASE TREATMENT ASSOCIATED WITH NON-INSULIN DEPENDENT DIABETES MELLITUS (NIDDM)

I.V. Medvedeva, E.E Dorodneva, M.A. Ushakova, E.O. Gerasimenko, E.A. Butina. Tyumen medical Academy, Tyumen, Russia

One of the promising methods of coronary heart disease (CHD) treatment Associated with NIDDM is the correction measures which decrease the cholesterol level in plasma and cellular structures, in particular dietary therapy.

Metods: We applied phospholipid product Vitol which is characterized by high content of choline fraction as well as exta'eme low content of lysoforms that determines it s hypocholesterolemic and antioxidant properties. 67 patients with CHD Associated with NIDDM (men aged 52 on average) were examined. Vitol WAS administered a daily dose equal to 5 g for 21 days together with antianginous remedies.

Results: The tests and clinical examinations exhibit the fact that phos- pholipid concentrate Vitol effects significantly upon patients with CHD, stable angina on exertion Associated with NIDDM with no ghost effect. The characteristic feature of this very remedy is it s structural influence at the level of platelet membrane which results in decrease of ratio total cholesterol/phospholipids in blood platelet membranes in patients with CHD Associated with NIDDM as well in decrease of oxidizing metabolism. All the above significantly improves functional properties of thrombocytes.

OBJECTIFICATION OF THERAPY EFFICIENCY "THE HEART AFTER INFARCTION": BETA-BLOCKERS AGAINST CALCIUM ANTAGONISTS

V. Tashchuk, O. Polyanska, R. Chaplinsky, T. Kulyk, A. EI-Telbany. Bukooinian Medical Academy, Chernivtsi, Ukraine

With the purpose of objectification of efficiency of therapy with beta- blockers ( I-st group) and calcium antagonists - Nifedipine (2-nd group) and Verapamil (3-d group) we investigated 58 patients in 2 months, half a year and I year after acute myocardial infarction. All patients were provided with Echocardiography, Stress Tests (transesophageal electrncardiostimulation (TEES)) and bicycle ergnmetry (BE), holter monitoring (HM) on the therapy background with nitrates, aspirin and angiotensin converting inhibitors. It was determined, accordingly, in the I, 2, 3, groups the redaction of total cholesterol (-3.0%, p > 0.5; -26.2%, p < 0.02; -2.8%, p > 0.5), the increase of ejection fraction (+3.8%, p < 0.05; +2.8%, p~< 0.05; +3.2%, p < 0.05), treshold loading under BE (+17.7%, p < 0.02; +27.4%, p < 0.02; +28.0%, p < 0.02) and frequency of TEES stop (+10.7%, p < 0.01; +6.7%, p < 0.01; +4.2%, p < 0.05), the decreasing of silent ishemia probability under HM (-20.7%, p, 0.05; -23.5%, p < 0.05; -53.4%, p < 0.001). Thus group of beta-blockers is assisted with the greatest increasing of TEES stop frequency, nifedipine group is connected with the maximal reduction of cholesterol level and threshold loading under BE; verapamil group caused the maximal reduction of silent ishemia in patients of remote period (half a year) after myocardial infarction.

THE TOXICOLOGICAL CHARACTERISTIC OF A NEW UKRAINIAN HYPOLIPIDEMIC DRUG UFIBRATE

S.A. Oliynik, N.A. Gorchakova, S.O. Gavriliuk, V.A. Tumanov, P.S. Nazar. National Medical University, Kiev, Ukraine

The primary toxicological characteristic of a new Ukrainian hypolipidemic drug - uflbrate - is given. By researches of acute toxicity of a drug on white mice, rats and cats is established, that it concerns to low-toxic substances. By long (6-month's) researches is established, that ufibrate in conditional- therapeutical and in conditional-toxic dozes did not cause changes a picture of blood at rabbits and rats. The clinical supervision which have been carried out on the patients with ischaemic disease of heart, have confirmed non-toxic characteristic of a drug. So, after trea~nent (reception of ufibrate tablets on 0.05 g three times per day after meal within 30 days) were not marked changes of a picture of blood and collateral reactions (including allergic). The presence of positive influence of ufibrate on some param- eters of a functional condition of a liver, in particular, authentic changes of parameters aspartate- and alanintransaminases, lactatdehydrogenase and cholineaesterase activities, is established. There were no one case of revealed deterioration of functional tests o f a liver and of nitrogenium excreting function of kidneys. The patients well transferred a combination of ufibrate with antianginal drugs and antagonists o f calcium channels.

71st EAS Congress and Satellite Symposia

( f,