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Biomedical subjects

A Gil

Publications and source records attributed to A Gil.

At least 109 records · Page 6Linked to original sources
PubMed2000

High frequencies of telomeric associations, chromosome aberrations, and sister chromatid exchanges in ulcerative colitis.

OBJECTIVE: Chromosome instability provides a predisposing background to malignancy, contributing to the crucial genetic changes in multistep carcinogenesis. The aim of this work was to analyze chromosome instability in patients with ulcerative colitis (UC) to achieve a better understanding of the increased risk for colorectal cancer. METHODS: Peripheral blood lymphocyte cultures from 20 untreated UC patients and 24 controls were used to study chromosome instability by assessing telomeric associations (TAS), chromosome aberrations (CA), and sister chromatid exchanges (SCE). RESULTS: Mean frequencies of TAS and CA were significantly increased in UC patients compared to controls (p < 0.001). Chromosomes 10, 11, 21, 16, and 19 were the most frequently involved in TAS. A total of 104 CA clustered in 66 breakpoints could be exactly localized. Seven nonrandom bands significantly affected in UC patients were found (p < 0.004), showing a significant correlation with the location of cancer breakpoints (p < 0.003), particularly with colorectal carcinoma rearrangements. SCE analysis showed higher levels in patients compared to controls (p < 0.006), but no differences were observed in cell cycle kinetics. CONCLUSIONS: Our results demonstrate the presence of an unstable genome in UC patients that could be related to the cancer development observed in this disease.

Adolescent↗
PubMed2000

Dietary trans fatty acids affect docosahexaenoic acid concentrations in plasma and liver but not brain of pregnant and fetal rats.

The aim of the present study was to investigate the maternal-fetal transport, incorporation, and effects on liver delta-6 fatty-acid desaturase activity of dietary trans fatty acids in pregnant rats. Three groups of six rats each were fed three experimental diets containing approximately 0%, 15%, and 30% of trans fatty acids but containing the same proportion of linoleic (18:2 n-6) and a-linolenic (18:3 n-3) acids for 10 wk. On d 20 of pregnancy, the animals from each group were killed. We determined the fatty acid profiles in plasma, brain, and liver microsomes of pregnant rats, as well as in placenta and fetal liver and brain. No changes were found in the number of fetuses of the pregnant rats. Trans fatty acids were incorporated in high concentrations in placenta and in maternal and fetal tissues, except brain, strongly elevating the linoleic acid proportion and lowering that of docosahexaenoic acid. The delta-6 fatty-acid desaturase activity in the liver microsomes of the pregnant rats was inhibited by trans isomers. In conclusion, high intakes of trans fatty acids partially inhibit liver delta-6 fatty-acid desaturase in pregnant rats, which may explain, in part, the low concentrations of docosahexaenoic acid in pregnant and fetal tissues. However, the fatty acid composition of both fetal and pregnant rat brain remains mostly unaffected regardless of the dietary trans fatty acid content.

Animals↗
PubMed2000

Potential use of spectral image analysis for the quantitative evaluation of estrogen receptors in breast cancer.

Evaluation of estrogen receptor (ER) content is an important factor in the choice of therapy and prognosis of breast cancer patients. In this study, we demonstrate a new spectral image analysis technique for objective and quantitative evaluations of stained specimens. The SpectraCube system was used to analyze nuclear antigens in thirteen cases of breast cancer stained by the immunoperoxidase method with hematoxylin counterstain. Spectral imaging segregated the spectrum of diaminobenzidine (DAB) from the background color of hematoxylin and a spectral index was calculated. The spectral index essentially agreed with the pathologist's index (on a scale of 0 to 3) in seven out of the thirteen cases. A substantial number of ER positive pixels was detected in the two cases scored as 0 by the pathologist's index. In a test case scored as 1 by the pathologist's index we detected a significant number of pixels, representing 47% of the nuclei, with DAB-intensity values higher than the cut-off value of 1.2. These data suggest that spectral image analysis is a sensitive method providing intensive information with high reproducibility. Our spectral imaging method is highly flexible, enabling the user to define the spatial resolution of the analyzed specimen by choosing the number of pixels per one nucleus.

3,3'-Diaminobenzidine↗
PubMed1999

A single amino acid near the C terminus of the synaptosomeassociated protein of 25 kDa (SNAP-25) is essential for exocytosis in chromaffin cells.

Amperometry in chromaffin cells expressing green fluorescent protein (GFP) fused to synaptosome-associated protein of 25 kDa (SNAP-25) have been used to test the involvement of single amino acids in exocytotic function, overcoming some of the limitations of studies based on Botulinum neurotoxin cleavage, as this occurs at defined sites of the protein. Constructs containing either the whole SNAP-25 polypeptide or several deleted forms lacking its C-terminal domain were heavily overexpressed in transfected cells. All GFP-fusions were located in both the cytoplasm and the plasma membrane. Although a construct containing complete SNAP-25 sustained normal secretion, removal of four or more amino acids of its C terminus greatly altered the overall rate and extent of exocytosis. Further mutational analysis proved that Leu203, the fourth residue from the C terminus, is critical for secretion. Kinetics of single granule fusions from cells expressing truncated forms showed slow onset and decay times when compared with control cells expressing full SNAP-25. Thus, these data provide direct evidence for the involvement of a specific residue of SNAP-25 in exocytosis and show that overexpression of GFP-SNAP contructs combined with single vesicle fusion measurements constitutes a powerful approach to dissect the structural elements playing a role in individual steps of the exocytotic cascade.

Animals↗
PubMed1999

Signal to noise analysis of multiple color fluorescence imaging microscopy.

BACKGROUND: Various approaches that were recently developed demonstrate the ability to simultaneously detect all human (or other species) chromosomes by using combinatorial labeling and fluorescence in situ hybridization (FISH). With the growing interest in this field, it is important to develop tools for optimizing and estimating the accuracy of different experimental methods. METHODS: We have analyzed the principles of multiple color fluorescence imaging microscopy. First, formalism based on the physical principles of fluorescence microscopy and noise analysis is introduced. Next, a signal to noise (S/N) analysis is performed and summarized in a simple accuracy criterion. The analysis assumes shot noise to be the dominant source of noise. RESULTS: The accuracy criterion was used to calculate the S/N of multicolor FISH (M-FISH), spectral karyotyping, ratio imaging, and a method based on using a set of broad band filters. Spectral karyotyping is tested on various types of samples and shows accurate classifications. We have also tested classification accuracy as a function of total measurement time. CONCLUSIONS: The accuracy criterion that we have developed can be used for optimizing and analyzing different multiple color fluorescence microscopy methods. The assumption that shot noise is dominant in these measurements is supported by our measurements.

Female↗
PubMed1999

Effect on platelet FXIII and partial characterization of Lonomin V, a proteolytic enzyme from Lonomia achelous caterpillars.

Contact with Lonomia achelous caterpillars venom induces a severe bleeding syndrome in humans. A constant finding in all reported cases is a marked decrease of blood coagulation factor XIII (FXIII), which has been attributed to the presence of a proteolytic enzyme, isolated and named Lonomin V, in the hemolymph and hair secretion. In this study, the effect of Lonomin V on transglutaminase activity from human plasma, rabbit plasma, and platelet FXIII was analyzed. The decrease of activity was more pronounced in platelet (A2) when compared with rabbit plasma (AB) and human plasma FXIII (A2B2). This finding might be explained by the differences in FXIII molecular structure. In addition, platelet FXIII molecule was degraded by Lonomin to several fragments of low molecular mass. Lonomin V was stable over a wide range of pH (6-8.5) and temperatures of -70 degrees C, -20 degrees C and between 4 to 24 degrees C, with a progressive decrease at 37 degrees C and total inactivation at 60 degrees C after 2 hours incubation. Diisopropyl fluoro-phosphate, phenylmethylsulfonyl fluoride, tosyl-l-lysine chloromethyl ketone, and iodoacetamide abolished the effect of Lonomin V on FXIII; in contrast dithiothreitol and EDTA-Na enhance the activity. We concluded that Lonomin V is a serine proteinase with a free Cys essential for the enzymatic activity. Due to its proteolytic activity on FXIII, with concomitant impairment of fibrin cross-linking, Lonomin V might be useful in association with thrombolytic drugs for preventing rethrombosis.

Animals↗
PubMed1999

Whole cell lysate enzyme immunoassays vs. recombinant glycoprotein G2-based immunoassays for HSV-2 seroprevalence studies.

Seroepidemiology studies of herpes simplex virus type 2 (HSV-2) infections have been difficult to carry out because antibodies to HSV type 1 (HSV-1) show an extensive cross-reactivity with HSV-2 antigens. Many kits available currently are not entirely type specific for serodiagnosis of HSV-2 infections and therefore do not allow reliable discrimination of past exposure to these closely related alphaherpes viruses. Attempts to develop type-specific antigens have focused on the envelope glycoproteins, particularly glycoprotein G (gG). A cross-sectional study was carried out to examine the seroprevalence of antibodies to HSV-2 among healthy university students, using different methods: a whole cell lysate enzyme-linked immunosorbent assay (ELISA), two different ELISAs, and a newly developed immunoblot assay, the last three based on recombinant gG2. HSV-2 prevalence was 24 times higher with the whole cell lysate ELISA (31%; 95% confidence interval [CI]: 27-35%) than the ELISAs and the immunoblot assay based on recombinant gG2 (1.3%; 95% CI: 0.1-2.5%), thus showing the inaccuracy of commercial tests based on whole-antigen preparations for epidemiological studies. Laboratories should be cautious and ensure that commercial tests for HSV typing are based on type-specific glycoproteins.

Adolescent↗
PubMed1999

Influence of dietary nucleotides on plasma immunoglobulin levels and lymphocyte subsets of preterm infants.

We examined the effects of nucleotide supplementation to a preterm adapted milk formula on the lymphocyte subsets and plasma IgG, IgM and IgA levels in preterm infants for the first three months of life. Two groups of preterm infants received a milk formula or the same formula supplemented with CMP, AMP, UMP, GMP and IMP to mimic the concentration of acid-soluble nucleotides found in human milk. Blood samples were obtained at birth, 10 days, 20-30 days and 3 months of age. Preterm infants fed the nucleotide formula exhibited higher plasma levels of IgM in all postnatal study periods than neonates fed the standard formula; moreover, IgA was also higher at 3 months of age in nucleotide formula fed infants. No major differences were seen between groups for IgG levels and lymphocyte subsets. Thus, dietary nucleotides appear to exert actions on immature human neonate lymphocytes enhancing the in vivo production of Ig which may have a role in the defense capacity of neonates.

Aging↗
PubMed1999

Addition of vitamin E to long-chain polyunsaturated fatty acid-enriched diets protects neonatal tissue lipids against peroxidation in rats.

BACKGROUND: Tissue 10:4(n-6) and 22:6(n-3) status have been correlated with neonatal development and growth. Artificial formulas for neonates have been supplemented with long chain polyunsaturated fatty acids (LCP) from animal and marine sources which may enhance sensitivity of cellular membranes to oxidative damage. Diet-derived antioxidants like vitamin E play a key role in the protection of tissue lipids against oxidation. AIM OF THE STUDY: We seek to determine the influence of dietary vitamin E on tissue sensitivity to oxidative stress in rats fed for 4 weeks on diets enriched in (n-3) and (n-6) long-chain polyunsaturated fatty acids. METHODS: Weanling rats received 10% fat diets that provided 18:1(n-9), 18:2(n-6) and 18:3(n-3) in a similar ratio to that of rat milk (group A), supplemented with fish oil (groups B and B+E) and supplemented with (n-6) and (n-3) LCP from an animal phospholipid concentrate (groups C and C+E). Vitamin E (500 mg vitamin E/kg fat) was added to diets B+E and C+E. Tissue fatty acid content and the activities of catalase, superoxide dismutase, glutathione transferase und glutathione peroxidase in liver and brain were measured. Glutathione status, vitamin E and the production of thiobarbituric acid reactive substances (TBARs) after incubation of erythrocyte, liver and brain lipids with inducers of enzymatic or non-enzymatic lipid peroxidation was measured. RESULTS: Group B registered significantly lower total superoxide dismutase activity than group B+. Catalase activity was significantly higher in group C than in group C+E. Hepatic total and reduced glutathione levels were decreased in vitamin E supplemented groups compared to unsupplemented ones. TBARs production in erythrocyte lipids was significantly higher in groups B and C compared to vitamin E supplemented groups B+E and C+E. CONCLUSIONS: This study shows that the addition of vitamin E protected erythrocyte and liver microsome lipids enriched in (n-3) and (n-6) LCP from lipid peroxidation during the postnatal development of rats. The protection was more effectively in group C+E than in group B+E.

Animals↗
PubMed1999

Positional analysis of triglycerides and phospholipids rich in long-chain polyunsaturated fatty acids.

Four sources of long-chain polyunsaturated fatty acids (LCP) differing in their chemical structure (triglycerides or phospholipids) and in their origin (tuna triglycerides, fungal triglycerides, egg phospholipids, and pig brain phospholipids) were analyzed to determine the distribution of the component fatty acids within the molecule. Lipase and phospholipase A2 hydrolysis was performed to obtain 2-monoacylglycerols and lysophospholipids, respectively, which allowed us to determine the distribution of fatty acids between the sn-2 and sn-1,3 positions of triglycerides or between the sn-1 and sn-2 position of phospholipids. Fatty acids in the LCP sources analyzed were not randomly distributed. In tuna triglycerides, half of the total amount of 22:6n-3 was located at the sn-2 position (49.52%). In fungal triglycerides, 16:0 and 18:0 were esterified to the sn-1,3 (92.22% and 91.91%, respectively)18:1 and 18:2 to the sn-2 position (59.77% and 62.62%, respectively), and 45% of 20:3n-6 and only 21.64% of 20:4n-6 were found at the sn-2 position. In the lipid sources containing phospholipids, LCP were mainly esterified to the phosphatidylethanolamine fraction. In egg phospholipids, most of 20:4n-6 (5.50%, sn-2 vs. 0.91%, sn-1) and 22:6n-3 (2.89 vs. 0.28%) were located at the sn-2 position. In pig brain phospholipids, 22:6n-3 was also esterified to the sn-2 (13.20 vs. 0.27%), whereas 20:4n-6 was distributed between the two positions (12.35 vs. 5.86%). These results show a different fatty acid composition and distribution of dietary LCP sources, which may affect the absorption, distribution, and tissue uptake of LCP, and should be taken into account when supplementing infant formulas.

Animals↗
PubMed1999

Oral administration of a turmeric extract inhibits LDL oxidation and has hypocholesterolemic effects in rabbits with experimental atherosclerosis.

The oxidation of low-density lipoproteins (LDL) plays an important role in the development of atherosclerosis. Curcumin is a yellow pigment obtained from rhizomes of Curcuma longa and is commonly used as a spice and food colouring. Curcumin and turmeric extracts have several pharmacological effects including antitumour, anti-inflammatory, antioxidant and antiinfectious activities although the precise mechanisms involved remain to be elicited. We evaluated the effect of an ethanol-aqueous extract obtained from rhizomes of C. longa on LDL oxidation susceptibility and plasma lipids in atherosclerotic rabbits. A total of 18 rabbits were fed for 7 weeks on a diet containing 95.7% standard chow, 3% lard and 1. 3% cholesterol, to induce atherosclerosis. The rabbits were divided into groups, two of which were also orally treated with turmeric extract at doses of 1.66 (group A) and 3.2 (group B) mg/kg body weight, respectively. A third group (group C) acted as a control. Plasma and LDL lipid composition, plasma alpha-tocopherol, plasma retinol, LDL TBARS, LDL lipid hydroperoxides and analysis of aortic atherosclerotic lesions were assayed. The low but not the high dosage decreased the susceptibility of LDL to lipid peroxidation. Both doses had lower levels of total plasma cholesterol than the control group. Moreover, the lower dosage had lower levels of cholesterol, phospholipids and triglycerides in LDL than the 3.2-mg dosage. In conclusion, the use of this extract could be useful in the management of cardiovascular disease in which atherosclerosis is important.

Administration, Oral↗
PubMed1999

Prevalence of hepatitis A in an institution for the mentally retarded in an intermediate endemicity area: influence of age length of institutionalization.

OBJECTIVE: This study aimed to examine the prevalence of antibodies against hepatitis A (anti-HAV) in a population of institutionalized mentally retarded persons compared with that of institutionalized non-mentally retarded persons in an area with moderate endemicity of HAV infection. METHODS: The study population was a group of mentally retarded persons aged between 10 and 30 years, institutionalized in one residence in Madrid, Spain. A group of non-mentally retarded subjects in the same age range was chosen as controls. Information about demography and duration of institutionalization was obtained by the investigator. Four drops of whole blood were obtained from each person by finger-stick, collected on filter paper, air-dried and stored at +4 degrees C until antibody determination. Eluates were tested by an ELISA method. RESULTS: A total of 314 institutionalized subjects were included: 157 mentally retarded (mean age and mean duration of institutionalization were 24.4+/-3 and 9.7+/-5 years respectively) and 157 non-mentally retarded (mean age and mean duration of institutionalization were 19.2+/-5 and 4.6+/-3 years respectively). The prevalence of anti-HAV antibody was 54% (95% confidence interval (CI) 50-58) in mentally retarded and 22% (95% CI 19-25) in non-mentally retarded [P<0.001; odds ratio (OR): 4.2 (95% CI 2.5-7)]. In both groups, these differences were not statistically significant for anti-HAV antibody prevalence between persons institutionalized for >5 years compared with those institutionalized for < or = 5 years. CONCLUSIONS: There results indicate that in an area of moderate endemicity the institutionalized mentally retarded are at increased risk of having acquired hepatitis A infection compared to the non-mentally retarded. In these institutionalized persons, regardless whether they are mentally retarded or not, prevalence is not influenced by age or length of stay in institutions.

Adolescent↗
PubMed1999

Lifestyle changes in free-living patients with peripheral vascular disease (Fontaine stage II) related to plasma and LDL lipid composition: a 15 month follow-up study.

Peripheral vascular disease (PVD) is characterized by arteriosclerosis and lower extremity ischemia which cause intermittent claudication. Patients grouped in the Fontaine stage II have more than 75% organic stenosis in their large coronary arteries and exhibit a number of alterations in blood coagulation and plasma lipids. The aim of this study was to evaluate an intervention program of lifestyle habits including dietary recommendations, moderate exercise and decreased smoking in a population of patients with PVD for a period of 15 months, with respect to plasma-lipid and lipoprotein composition as well as LDL susceptibility to peroxidation. These parameters are well known risk indicators of arteriosclerosis and coronary heart disease. A total 13 subjects diagnosed with PVD (Fontaine stage II) were selected, while a healthy age-matched group (n=20) was used as a reference. This study design was an uncontrolled trial of lifestyle interventions. The group of patients was examined at 0, 3, 6, 9, 12 and 15 months. Patients smoking one or more packets of cigarettes per day at the beginning of the study (54.2%) decreased smoking by as much as 7.7% 15 months later. In addition, physical activity intensified significantly (walking > 1 km: 13.1-77%) and treadmill running increased over the study period while the energy intake decreased by 10%. The percentage of saturated fat in the diet decreased by 10% while the intake of polyunsaturated fat rose, and monounsaturated-fat intake showed a parallel trend to increase; the average intake of cholesterol also fell by 10% and plasma triglycerides and HDL-cholesterol showed a trend to decrease and increase, respectively. No permanent changes in LDL lipid fractions for patients were detected during the follow-up period and no differences between patients and the age-matched reference group were found. The macrophage uptake of plasma-oxidized LDL was significantly higher in patients than in the reference group and no differences due to the intervention period were detected. In conclusion, the education in lifestyle and nutritional habits of patients with PVD led to reduced energy intake parallel with augmented physical activity as well to a fall in plasma triglycerides and a rise in HDL-cholesterol, which are good indicators of a reduced risk of vascular and myocardial complications.

Analysis of Variance↗
PubMed1999

Effect of extra-virgin olive oil and fish-oil supplementation on plasma lipids and susceptibility of low-density lipoprotein to oxidative alteration in free-living spanish male patients with peripheral vascular disease.

The aim of this study was to analyse the in vivo effect of a diet rich in extra-virgin olive oil and a fish-oil supplement on plasma and lipoprotein fatty-acid composition and on LDL susceptibility to oxidative modification in free-living Spanish male patients with peripheral vascular disease. A total of 12 patients were included in the experimental group which received extra virgin olive oil and a fish oil supplement (group OF). On the other hand, 13 patients which had refined olive oil as the main visible fat were considered as Control group. Plasma triglycerides decreased significantly after three months of dietary intervention. The LDL and plasma fatty-acid pattern in the group OF was characterized by a significant increase in n-3 polyunsaturated fatty acids, mainly 20:5 n-3 and 22:6 n-3. The slopes of LDL oxidative susceptibility were similar between baseline and endpoint values in both groups. However, the uptake of oxidized LDL by macrophages was significantly reduced in OF patients in comparison with the Control group. In conclusion, the daily intake of about 40 g/d of extra-virgin olive oil in combination with a daily supplement of 16 g of fish oil for 3 months in patients with peripheral vascular disease leads to a plasma-lipid profile less atherogenic than in patients having refined olive oil as the main visible food fat. The simultaneous consumption of alpha-tocopherol and natural antioxidants provided by extra-virgin olive oil seems to have a protective effect on the LDL susceptibility to oxidative modifications in spite of a higher proportion of n-3 polyunsaturated fatty acids.

Cholesterol, HDL↗
PubMed1999

Effects of dietary polyunsaturated fatty acids and nucleotides on tissue fatty acid profiles of rats with carbon tetrachloride-induced liver damage.

The deficiency of polyunsaturated fatty acids (PUFA) that occurs in plasma of patients with liver cirrhosis has been assessed in rats with severe steatosis and mild liver necrosis induced by repeated administration of low doses of carbon tetrachloride (CCl(4)). The contribution of both dietary (n-3) long-chain PUFA and nucleotides to the recovery of the altered fatty acid profiles of tissue lipids of these rats has also been studied. Two groups of rats were used. The first was intraperitoneally injected 0.15 ml of a 10% (v/v) CCl(4)solution in paraffin per 100 g of body weight, three times a week for 9 weeks; the second received paraffin alone. After the treatment, six rats of each group were killed. Afterwards, the remaining controls were fed a semipurified diet (SPD) for 3 weeks, and the remaining rats in the CCl(4)group were divided into three new groups: the first was fed the SP diet; the second was fed the SP diet supplemented with 1% (n-3) polyunsaturated fatty acids (PUFA diet); and the third was fed the SP diet supplemented with 250 mg nucleotides per 100 g diet (NT diet). Fatty acids of plasma, erythrocyte membranes and liver microsomes were analyzed. Decreases in linoleic and arachidonic acids in both total plasma lipids and liver microsomal phospholipids were the main findings due to CCl(4)treatment. The rats that received CCl(4)and the PUFA diet showed the lowest levels of (n-6) PUFA and the highest levels of (n-3) PUFA in liver microsomal phospholipids, as well as a significant increase of (n-3) PUFAs in total plasma lipids. The animals that received the NT diet showed no signs of fatty infiltration and exhibited the highest levels of (n-6) PUFAs in liver microsomal phospholipids. These results show that CCl(4)affects fatty acid metabolism which is accordingly reflected in altered tissue fatty acid profiles, and that balanced diets containing PUFA and nucleotides are important for the recovery of the damaged liver in rats.

Animals↗
PubMed1999

[The 4 Provinces Study: its principal objectives and design. The Researchers of the 4 Provinces Study].

INTRODUCTION AND OBJECTIVES: Spain shows an important variation in the geographical distribution of ischaemic heart disease and cerebrovascular disease mortality. In this article, the primary objectives and design of the Four Provinces study are described. In this study we analyzed the contribution of environmental factors (diet, lipid profile and plasma antioxidants), acting in childhood, to explain differences in cardiovascular mortality rates between different provinces in Spain. METHODS: An ecological design was projected in which the units to study were four Spanish provinces with a wide variation in cardiovascular mortality in adulthood. The design compares diet, anthropometric variables and biological markers (particularly plasma lipids and antioxidant levels) in 6-7-year-old children, between the two provinces with the highest cardiovascular mortality rates and the two provinces with the lowest. The information for each province is collected in a cross-sectional manner in a representative sample of children from each province. DISCUSSION: Evidence from the literature concerning Northern European countries suggest the contribution of environmental factors during early age in the development of cardiovascular disease in adulthood. The "Four Provinces" study will provide, for the first time, information about the influence of factors in early childhood of cardiovascular risk in a Mediterranean country. The study will also offer interesting data about food intake during school age in four provinces and it will allow us to estimate values of population of the variables of interest in those provinces.

Antioxidants↗
PubMed1999

Olive oil- and fish oil-enriched diets modify plasma lipids and susceptibility of LDL to oxidative modification in free-living male patients with peripheral vascular disease: the Spanish Nutrition Study.

The present study describes a clinical trial in which Spanish patients suffering from peripheral vascular disease (Fontaine stage II) were given specific lipid supplements. Designed as a longitudinal intervention study, patients were provided with olive oil for 3 months, followed by a 3 month wash-out period, then supplemented with a combination of fish oil and olive oil for the final 3 months. Changes in plasma and lipoprotein fatty acid composition and susceptibility of LDL to in vitro oxidation were examined. Furthermore, lipid-supplement-induced changes in LDL properties were measured as relative electrophoretic mobility and macrophage uptake. In addition, thirteen patients not provided with olive oil and fish oil were included as a control group and twenty healthy age-matched individuals were used as a reference group. A complete clinical study and a nutritional survey concerning food habits and lifestyle were performed every 3 months. Yao indices and claudicometry did not change significantly with dietary intervention although changes in plasma lipid composition suggested an improvement in the condition of the patients. The intake of the fish-oil supplement resulted in significantly increased plasma levels of eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) in comparison with baseline concentrations, olive-oil and control groups. Fish-oil consumption significantly decreased plasma triacylglycerol levels compared with the olive-oil period, control and reference groups. The susceptibility of LDL to Cu-mediated oxidation was lower in the patients consuming olive oil and the fish-oil supplement than in the control group, and the uptake of LDL by macrophages was significantly lower in the group supplemented with fish oil. In conclusion, consumption of olive oil together with a dietary supplement of fish oil may be useful in the nutritional management of patients suffering from peripheral vascular disease in terms of increasing plasma n-3 long-chain polyunsaturated fatty acids and decreasing susceptibility of LDL to oxidation.

Cholesterol, LDL↗