FO Groups Research Articles

Unsaturated Fatty Acids in Fish Oil Play a Role in Adequate Fat Distribution to Plasma, Liver and White Adipose Tissue

The components bringing the effects of fish oil on glucose and lipid metabolism are unclear. We used hydrogenated fish oil, which has saturated fatty acids with the same carbon chain lengths as the unsaturated fatty acids in fish oil, to clarify the functions of these unsaturated fatty acids on the improvements in lipid and glucose metabolism in mice. Mice divided into 3 groups were fed different diets: fish oil diet (FO), hydrogenated fish oil diet (HFO), and soybean oil diet (SBO) as a control. Body weight gain and white adipose tissue weight in the HFO and FO groups were significantly decreased compared with those in the SBO group. However, in the HFO group, the triglyceride (TG) levels in plasma were significantly decreased, while the lipids levels in the liver were remarkably increased compared with those in the FO group. Regarding the fatty acid composition in the liver and white adipose tissue in the HFO group, in parallel with the up-regulation of stearoyl-CoA desaturase 1 mRNA, relative amounts of C16 : 1 and C18 : 1 were significantly increased. By contrast, blood glucose levels in the oral glucose tolerance test had not deteriorated in the HFO group. Our results indicate that unsaturated fatty acids in the FO diet decrease lipid levels in the liver and maintain the balance of lipid levels in plasma, liver and white adipose tissue; in addition, in the HFO group, C16 : 1 and C18 : 1 synthesized in the liver and white adipose tissue may improve glucose tolerance and lipid metabolism.

Effects of High Dose Fish Oil Supplementation on Delayed Onset Muscle Soreness and Inflammatory Markers

Delayed muscle soreness (DOMS) is caused, in part, by muscle damage associated with the eccentric component of exercise and this muscle damage can result in an inflammatory response by the body. Fish oil rich in eicosapaetonic acid (EPA) and docosahexeoric acid (DHA) have been shown to have anti-inflammatory properties. However, studies have not investigated the impact of consuming a high dose offish oil for a prolonged period of time on DOMS. The purpose of this study was to determine the effects of high dose fish oil supplementation on DOMS and the associated muscle inflammatory response. The control group (CON; n=10) consumed 3.45 grams of wheat germ and the experimental group (FO; n=10) received 3.0 grams fish oil per day for 65 days. On Day 60, participants performed eccentric muscle contractions at an angular velocity of 60°-sec -1 through 20° to 90° of knee flexion until exhaustion. Perceived muscular soreness, isometric strength, and muscle inflammatory markers were measured before exercise and at 8, 24,48, 72, 96 and 120 hours post-exercise. We found no significant differences between the CON and FO groups for all variables. The data suggests the fish oil had no effect on inflammatory markers and the signs and symptoms of DOMS when compared to a placebo.

The peroxynitrite evoked contractile depression can be partially reversed by antioxidants in human cardiomyocytes

In this study, we aimed to determine the contribution of peroxynitrite-dependent sulfhydryl group (SH) oxidation to the contractile dysfunction in permeabilized left ventricular human cardiomyocytes using a comparative approach with the SH-oxidant 2,2'-dithiodipyridine (DTDP). Additionally, different antioxidants: dithiothreitol (DTT), reduced glutathione (GSH) or N-acetyl-L-cysteine (NAC) were employed to test reversibility. Maximal isometric active force production (F(o)) and the maximal turnover rate of the cross-bridge cycle (k(tr,max)) illustrated cardiomyocyte mechanics. SH oxidation was monitored by a semi-quantitative Ellman's assay and by SH-specific protein biotinylation. Both peroxynitrite and DTDP diminished F(o) in a concentration-dependent manner (EC(50,peroxynitrite) = 49 microM; EC(50,DTDP) = 2.75 mM). However, k(tr,max) was decreased only by 2.5-mM DTDP, but not by 50 microM peroxynitrite. The diminution of F(o) to zero by DTDP was paralleled by the complete elimination of the free SH groups, while the peroxynitrite-induced maximal reduction in free SH groups was only to 58 +/- 6% of the control (100%). The diminutions in F(o) and free SH groups evoked by 2.5-mM DTDP were completely reverted by DTT. In contrast, DTT induced only a partial restoration in F(o) (DeltaF(o,): approximately 13%; P < 0.05) despite full reversion in protein SH content after 50 microM peroxynitrite. Although, NAC or DTT were equally effective on F(o) after peroxynitrite exposures, NAC or GSH did not restore F(o) or k(tr,max) after DTDP treatments. Our results revealed that the peroxynitrite-evoked cardiomyocyte dysfunction has a small, but significant component resulting from reversible SH oxidation, and thereby illustrated the potential benefit of antioxidants during cardiac pathologies with excess peroxynitrite production.

Effects of Yogurt Supplemented with Fish Oil on Plasma Lipid and Glucose Concentrations, and Liver Lipid Contents in Mice

The aim of this study is to clarify the effects of yogurt supplemented with fish oil on plasma lipid and glucose concentrations, and hepatic lipid contents in mice. Male Crlj:CD-1 (ICR) mice were fed five experimental diets for 12 weeks. The experimental diets were as follows: without yogurt and fish oil (control diet); 10% (w/w) yogurt without fish oil [10% FO(-)]; 10% yogurt with fish oil [10% FO(+)]; 30% yogurt without fish oil [30% FO(-)]; 30% yogurt with fish oil [30% FO(+)]. Plasma triacylglycerol concentrations in the 10% FO(+) and 30% FO(-) groups were significantly lower than that in the control diet group (p < 0.05 and p < 0.05, respectively). Plasma total cholesterol and phospholipid concentrations were significantly lower in the 30% FO(+) group than in the control diet group (p < 0.005). Concentrations tended to be lower with supplementation with fish oil. Plasma glucose concentrations in the 10% FO(+) and 30% FO(+) groups were significantly lower than those in the control diet group (p < 0.005 and p < 0.01, respectively). Hepatic triacylglycerol and total cholesterol contents in the 30% FO(+) group were significantly lower than those in the control diet group (p < 0.05 and p < 0.005, respectively). These results suggest that plasma triacylglycerol and glucose concentrations are effectively decreased by supplementation of yogurt with fish oil.

Correlation between changes in blood fatty acid composition and visual sustained attention performance in children with inattention: effect of dietary n−3 fatty acids containing phospholipids

BackgroundIncreasing evidence supports n−3 fatty acid (FA) supplementation for patients with psychiatric disorders, such as attention deficit hyperactivity disorder. However, the exact metabolic fate of dietary eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on different glyceride carriers remains unclear. ObjectiveWe investigated whether conjugation of EPA and DHA to phospholipid (PL-n−3) or to triacylglycerol (fish oil; FO) affects their incorporation in blood compartments and influences executive functioning. DesignChildren aged 8–13 y with impaired visual sustained attention performance received placebo, 250 mg/d EPA + DHA esterified to PL-n−3 (300 mg/d phosphatidylserine), or FO for 3 mo in a randomized double-blind manner. Main outcome measures included plasma and erythrocyte FA profile and continuous performance test results (Test of Variables of Attention; TOVA). ResultsSixty of the 83 children enrolled completed the interventions (n = 18–21 per group). There was an enrichment of EPA (1.5–2.2-fold), docosapentaenoic acid (DPA; 1.2-fold), and DHA (1.3-fold) in the PL fraction in the plasma of FO- and PL-n−3–fed children. In erythrocytes, only PL-n−3 resulted in a significant reduction (≈30%) of very-long-chain saturated FAs (C20-24) and in an increase (1.2- and 2.2-fold, respectively) in linoleic acid and DPA. Total TOVA scores increased in the PL-n−3 (x̄ ± SD: 3.35 ± 1.86) and FO (1.72 ± 1.67) groups but not in the placebo group (−0.42 ± 2.51) (PL-n−3 > FO > placebo; P < 0.001). A significant correlation between the alterations in FAs and increased TOVA scores mainly occurred in the PL-n−3 group. ConclusionConsumption of EPA+DHA esterified to different carriers had different effects on the incorporation of these FAs in blood fractions and on the visual sustained attention performance in children. This trial was registered at clinicaltrials.gov as NCT00382616.

Maternal fish oil supplementation benefits programmed offspring from rat dams fed low-protein diet

We hypothesized that maternal fish oil supplementation would prevent the programming of offspring cardiovascular alterations. Forty rats were fed during the perinatal period with normal-protein (NP) diet (19%) or low-protein (LP) diet (5%) and also received daily fish oil supplement (Fo). Offspring were divided into male and female NP, NP plus Fo, and LP and LP plus Fo groups (n = 5, each) until 6 months old. Cardiac structure was examined. Both LP sexes had mild hypertension, but Fo supplementation minimized hypertension. The left ventricle was thicker in the LP groups and less thick in the LP plus Fo groups. The male LP had significantly fewer intramyocardial microcirculation, whereas the male LP plus Fo had 90% more microcirculation than the LP group. The LP group had significantly higher interstitial fibrosis, compared with the NP groups. Postnatal hypertension and collateral cardiovascular adverse remodeling were programmed by perinatal LP diet, whose outcomes were alleviated by maternal fish oil supplementation.

Fish oil infusion reverses 5-fluorouracil-induced impairments in mucosal immunity in mice

Anticancer drugs frequently have deleterious effects on host defense against infection, limiting their clinical application. We previously demonstrated continuous infusion of 5-fluorouracil (5FU) to reduce gut associated lymphoid tissue (GALT) mass and secretory IgA levels. This study was designed to examine the effects of concomitant infusion of fish oil on gut mucosal immunity in mice receiving 5FU. Male ICR mice were randomized to the control (n=12), 5FU (n=12), or 5FU+FO (n=10) group. The 5FU and 5FU+FO groups received continuous IV infusion of 5FU at 10 mg/kg for 5 days. The 5FU+FO group was given a simultaneous infusion of 10 ml/kg of a 10% fish oil emulsion. The controls received normal saline at 0.3 ml/h. During these treatments, all mice were allowed free access to chow and water ad libitum. Then, the mice were sacrificed and GALT lymphocytes were isolated from Peyer's patches (PPs), the intraepithelial space (IE), and the lamina propria (LP). Small intestinal, nasal and broncho-alveolar (BALF) washings were also obtained. Lymphocyte yields from each site and phenotypes (CD4, CD8, alphabetaTCR, gammadeltaTCR, B220) were determined. IgA levels in the washings were measured with ELISA. The 5FU group had significantly lower IE and LP lymphocyte numbers and small intestinal and BALF IgA levels than the control group, with no differences in the percentages of any phenotypes. However, fish oil infusion restored IE and LP lymphocyte numbers and BALF IgA to control group levels. Fish oil infusion along with 5FU preserves GALT lymphocyte numbers and respiratory IgA levels.

Dietary fatty acids modulate chronic colitis, colitis-associated colon neoplasia and COX-2 expression in IL-10 knockout mice

Objective The effects of different dietary oils on the development of colitis-associated colon cancer have not been studied. The present study examined the effect of different dietary oils on the severity of chronic colitis, development of colitis-associated premalignant changes, and colonic expression of cyclooxygenase-2 (COX-2) in interleukin-10 knockout (IL-10 −/−) mice. Methods IL-10 −/− mice were fed chow supplemented with corn oil (CO; control, n = 28), olive oil (OO; n = 29), or fish oil (FO; n = 35) for 12 wk and their colons were studied for colitis score, premalignant changes, and COX-2 expression. Results The average colitis score was higher in the FO than in the CO group. Similarly, the incidence of severe colitis (score ≥ 3) was significantly higher in the FO than in the CO and OO groups (50% versus 7.7% and 3.7%, respectively, P < 0.05). Dysplasia was more frequent in the FO and less frequent in the OO than in the CO group (47% and 4% versus 15%, respectively, P < 0.05). Conversely, aberrant crypt foci and crypt index were significantly higher in the FO than in the CO group. Colitis score, aberrant crypt foci, and crypt index did not differ between the OO and CO groups. COX-2 immunostaining was significantly lower in the OO than in CO group ( P < 0.05) but not different between the FO and CO groups. Conclusions In IL-10 −/− mice, fish oil exacerbates chronic colitis and colitis-associated premalignant changes. Conversely, olive oil inhibits COX-2 immunostaining and decreases the risk of neoplasia associated with chronic colitis.

The inhibitory effect of flaxseed on the growth and metastasisof estrogen receptor negative human breast cancer xenograftsis attributed to both its lignan and oil components

Our previous studies have shown that dietary flaxseed (FS) can reduce the growth and metastasis of human estrogen receptor negative (ER-) breast cancer in nude mice. The aims of our study were to determine (i) whether the tumor inhibitory effect of FS was due to its oil (FO), lignan secoisolariciresinol diglycoside (SDG), or both components, and (ii) whether the effect on tumor growth was related to increased lipid peroxidation. Athymic nude mice were orthotopically injected with ER- breast cancer cells (MDA-MB-435) and 8 weeks later were fed either the basal diet (BD) or BD supplemented with 10% FS, SDG, FO, or combined SDG and FO (SDG + FO) for 6 weeks. The SDG and FO levels were equivalent to the amounts in the 10% FS. Compared to the BD group, the tumor growth rate was significantly lower (p < 0.05) in the FS, FO, and SDG + FO groups, in concordance with decreased cell proliferation and increased apoptosis; however, these did not significantly relate to the lipid peroxidation, indexed as malonaldehyde (MDA), in the primary tumors. Lung metastasis incidence was reduced (16-70%) by all treatments, significantly in the FS and SDG + FO groups. The distant lymph node metastasis was significantly decreased (52%) only in the FO group. Although the total metastasis incidence was lowered (42%) significantly only in the SDG + FO group, all treatment groups did not differ significantly. In conclusion, FS reduced the growth and metastasis of established ER- human breast cancer in part due to its lignan and FO components, and not to lipid peroxidation.

Lack of Apoptosis in Listeria monocytogenes -Infected Thymocytes from Mice Fed with Dietary Lipids

The potential action of certain fatty acids has been studied since the early 1970s. Numerous effects on immune system functions have been related to dietary lipid administration; therefore, several of them have been applied in the treatment of inflammatory disorders. Nevertheless, n-3 polyunsaturated fatty acids may affect host resistance to infectious diseases. In addition, several studies have demonstrated that certain fatty acids are involved in apoptosis induction. Here, we have examined the action of different dietary lipids on the promotion of apoptosis in thymocytes from mice fed with dietary lipids and infected with Listeria monocytogenes. Thus, L. monocytogenes promoted an important cytotoxic effect in all of the groups, but it did not increase the percentage of DNA fragmentation. Similarly, an important increase of caspase-3 activity was demonstrated in OO and FO groups, but infection with L. monocytogenes exerted an inhibitory effect. Finally, L. monocytogenes did not modify proteasome activity among groups fed with dietary lipids. On the basis of this preliminary study, we can state that the infection of thymocytes from mice fed with dietary lipids does not promote a synergistic effect in the induction of apoptosis. Hence, these results may partially serve to elucidate the immune mechanisms involved in cells from mice fed with dietary lipids in an infectious process.

Dietary fatty acid composition affects the repeat swimming performance of Atlantic salmon in seawater

Repeated critical swimming performance trials ( U crit) were performed on Atlantic salmon ( Salmo salar) to test the null hypothesis that the source of dietary lipids (fish-based, poultry-based, and plant-based) does not influence exercise and recovery performance. Four diets were prepared by extensively replacing supplemental lipid from anchovy oil (AO; 100% AO at 150 g/kg) with cold pressed flaxseed oil (FO; 25% AO, 75% FO), sunflower oil (SO; 25% AO, 75% SO), or poultry fat (PF; 25% AO, 75% PF). These diets had equivalent protein and energy concentrations, but due to the different supplemental lipid sources, varied widely in their fatty acid composition. Fish fed AO had a significantly higher ( P<0.05) first U crit (2.62±0.07 body lenght s −1) than those fed PF (2.22±0.12 body lenght s −1) that had low muscle ratios of n-3 highly unsaturated fatty acids (n-3 HUFA) to saturated fatty acids (SFA) and arachidonic acid (AA), and high levels of oleic acid. Fish in the FO and SO diet groups swam as well as AO-fed fish in both swimming trials. The performance of fish fed AO decreased significantly ( P<0.05) during the second swimming trial (i.e. U crit2/ U crit1=0.92±0.02). No significant differences occurred between diet groups for the second swim trial. There was a positive correlation between both n-3 HUFA/SFA and n-3 HUFA/AA ratios, and U crit1. A negative correlation was found between dietary AA and oleic acids, and U crit1. The present study suggests that low dietary n-3 HUFA/ SFA and n-3 HUFA/AA ratios may negatively affect swimming performance. The former possibly can be offset by increasing linoleic acid in the presence of nutritionally adequate n-3 HUFA (e.g. SO diet). Lipid supplements consisting largely of vegetable oils did not compromise fish cardiorespiratory physiology under the conditions of this study.

Prenatal serotonin depletion changes the behavioral response in the nociceptive formalin test in rats.

In the past years, the attention of researchers is attracted to studying the mechanisms of the development of the central nervous system during prenatal ontogeny, when its major structural and functional characteristics are formed. A substantiated hypothesis assumes that serotonin (5-HT) is involved in these processes. It is known that, at early stages of embryonic development, 5-HT serves as an inducer of cellular and tissue processes and later acquires the properties of a neurotransmitter [1]. Change in the 5-HT content in the prenatal period induces disturbances in neuroembryogenesis leading to abnormalities in the brain development, which may be retained in postnatal ontogeny [1, 2]. The role of 5-HT in the prenatal development of the nociceptive system, in which it acts first as a morphogen and later as a key neurotransmitter, remains to be understood. In this study, we attempted to estimate, using the standard nociceptive formalin test [3, 4], the effect of prenatal 5-HT depletion on the parameters of pain sensitivity of Wistar rats after birth. Formalin test is an adequate model of a prolonged tonic pain, whose modulation in CNS, unlike the modulation of acute phasic [5] pain, is determined by other, unknown, mechanisms and which allows objective quantitative estimation of pain sensitivity. Earlier, in experiments with 25-day-old rats, we described in detail the behavioral response to formalin injection [6] (which already at this age was represented by two phases characteristic of mature rats [7]). This study was performed using young rats of the same age. The 5-HT depletion was performed by injecting an aqueous solution of an inhibitor of the 5-HT synthesis, parachlorphenylalanine (pCPA; ICN, United States) at a dose of 400 mg/kg (2 ml) intraperitoneally to the rat on day 9 of pregnancy. The term of injection was selected taking into account the published data that the maximum effect of pCPA (the greatest decrease in the 5-HT content not only in the mother, but also in the embryo) is expressed with in two to three days, i.e., in the crucial period (day 11‐12) of the formation of the serotonergic system [1, 8]. In the study, we used the fo lowing groups of female and male rats of offspring: (1) intact females; (2) control females that were injected with saline during pregnancy; (3) females that were subjected to 5-HT depletion during pregnancy. At the age of 25 days, the rat was placed into a chamber (25 〈 25 〈 25 cm) with transparent walls, and its twophase behavioral response (TPBR) to the formalin injection (2.5%, 10 μ l) into the plantar pad of the left hind foot was studied. We recorded the number of flexions and shakes patterns (the spinal level) and the duration (in second) of the licking (the supraspinal level) of the injected foot during each minute, until the response was finished (for 60 min). We analyzed the intensity and duration (in minutes) of the first and second phases and the duration of the period of rest (an interphase interval during which specific responses are absent). The formalin injection caused the above specific

Perfluoroalcohols

Perfluorinated primary alcohol adducts with triethylamine were prepared from lower perfluorinated acid fluorides and triethylamine monohydrofluoride. They are stable compounds and may be used as a source of R FO groups in organic syntheses.

단식중인 흰쥐에서 지방섭취가 생존 대사에 미치는 영향

완전 단식군의 흰쥐(FO)와 단식중에 소기름(FT), 옥수수기름(FC), 들깨기름(FP)만을 각각 투여하고 비교하니 FO군은 대부분 6일까지 생존하고 FT, FC, FP군은 대부분 16일까지 생존하였다. 폐사 직전의 모든 흰쥐 체중은 -24.5～-25%로 감소되었다. 단식중 유지 섭취는 다소 생명을 연장하나 식이 유지 종류에 따른 생존기간과 체중 변화에 미치는 영향에는 괄목할 차이가 없었다. 간장과 신장 무게 변화도 체중변화와 유사한 양상을 보였다. FO군의 혈중 총 cholesterol 농도, 중성지방 농도, 혈당은 대폭 감소되고 GPT 활성값과 BUN 농도는 상당 수준으로 증가되었다. FC군과 FP군에서 유지의 n-6/n-3비율 혹은 불포화도 특성 차이가 총 cholesterol농도, 중성지방농도, 혈당농도, BUN농도, 단식 초기의 GPT활성도에 영향을 주지 못했다. 그러나 기아사 직전의 FP군에서 GPT활성도가 FT, FC군보다 높았다. 본 연구에서 결론은 기아 상태의 흰쥐에 유지 단독 섭취는 유지 종류에 관계없이 생존일수를 다소 비슷하게 연장하였고 n-6/n-3비율, 불포화도 특성차이가 전반적으로 단식중의 대사에 미치는 영향에서 별다른 차이가 없었음을 알 수 있었다.

Relevance of dietary lipids as modulators of immune functions in cells infected with Listeria monocytogenes.

Nutritional status may have significant importance for the immune system, and particularly, unsaturated fatty acids may serve as modulators of immune functions. Clinical and epidemiological studies have demonstrated that fatty acids are involved in the reduction of the inflammatory processes that occur in diseases characterized by an overactivation of the immune system. At the same time, an increase in susceptibility to infection has also been reported. The importance of immune system modulation by dietary lipids in the presence of an intracellular bacterial pathogen, such as Listeria monocytogenes, was evaluated in the present study. BALB/c mice were divided into four groups which were each fed a low-fat (2.5% by weight) diet, an olive oil (OO; 20% by weight) diet, a fish oil (FO; 20% by weight) diet, or a hydrogenated coconut oil (HCO; 20% by weight) diet for 4 weeks. In each group, lymphocye proliferation was measured, and a reduction in the stimulation index was observed in the FO and HCO groups. Cytotoxicity exerted by L. monocytogenes was increased in the groups fed diets containing OO and FO after 6 h of incubation with the bacterium. An important increase in the production of reactive oxygen species was found in the groups fed the HCO diet after 12 h of incubation with L. monocytogenes. Finally, invasion and adhesion factors were not modified substantially by the action of dietary lipids, although these factors were reduced in cells from mice fed an FO diet. These results underline the importance of several dietary lipids as biological modulators of immune functions and their crucial role in the alteration of host natural resistance.

Dietary Supplementation with γ-Linolenic Acid or Fish Oil Decreases T Lymphocyte Proliferation in Healthy Older Humans

Animal and human studies have shown that greatly increasing the amounts of flax seed oil [rich in the (n-3) polyunsaturated fatty acid (PUFA) alpha-linolenic acid (ALNA)] or fish oil [FO; rich in the long chain (n-3) PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] in the diet can decrease mitogen-stimulated lymphocyte proliferation. The objective of this study was to determine the effect of dietary supplementation with moderate levels of ALNA, gamma-linolenic acid (GLA), arachidonic acid (ARA), DHA or FO on the proliferation of mitogen-stimulated human peripheral blood mononuclear cells (PBMC) and on the production of cytokines by those cells. The study was randomized, placebo-controlled, double-blinded and parallel. Healthy subjects ages 55-75 y consumed nine capsules/d for 12 wk; the capsules contained placebo oil (an 80:20 mix of palm and sunflower seed oils) or blends of placebo oil with oils rich in ALNA, GLA, ARA or DHA or FO. Subjects in these groups consumed 2 g of ALNA or 770 mg of GLA or 680 mg of ARA or 720 mg of DHA or 1 g of EPA plus DHA (720 mg of EPA + 280 mg of DHA) daily from the capsules. Total fat intake from the capsules was 4 g/d. The fatty acid composition of PBMC phospholipids was significantly changed in the GLA, ARA, DHA and FO groups. Lymphocyte proliferation was not significantly affected by the placebo, ALNA, ARA or DHA treatments. GLA and FO caused a significant decrease (up to 65%) in lymphocyte proliferation. This decrease was partly reversed by 4 wk after stopping the supplementation. None of the treatments affected the production of interleukin-2 or interferon-gamma by PBMC and none of the treatments affected the number or proportion of T or B lymphocytes, helper or cytotoxic T lymphocytes or memory helper T lymphocytes in the circulation. We conclude that a moderate level GLA or EPA but not of other (n-6) or (n-3) PUFA can decrease lymphocyte proliferation but not production of interleukin-2 or interferon-gamma.

Effects of dietary polyunsaturated fatty acids and hepatic steatosis on the functioning of isolated working rat heart under normoxic conditions and during post-ischemic reperfusion.

The purpose of this study was to modify the amount of 22:4 n-6, 22:5 n-6 and 20:5 n-3 in cardiac phospholipids and to evaluate the influence of these changes on the functioning of working rat hearts and mitochondrial energy metabolism under normoxic conditions and during postischemic reperfusion. The animals were fed one of these four diets: (i) 10% sunflower seed oil (SSO); (ii) 10% SSO + 1% cholesterol; (iii) 5% fish oil (FO, EPAX 3000TG, Pronova) + 5% SSO; (iv) 5% FO + 5% SSO + 1% cholesterol. Feeding n-3 PUFA decreased n-6 PUFA and increased n-3 PUFA in plasma lipids. In the phospholipids of cardiac mitochondria, this dietary modification also induced a decrease in the n-6/n-3 PUFA ratio. Cholesterol feeding induced marked hepatic steatosis (HS) characterized by the whitish appearance of the liver. It also brought about marked changes in the fatty acid composition of plasma and mitochondrial phospholipids. These changes, characterized by the impairment of deltaS- and delta6-desaturases, were more obvious in the SSO-fed rats, probably because of the presence of the precursor of the n-6 family (linoleate) in the diet whereas the FO diet contained large amounts of eicosapentaenoic and docosahexaenoic acids. In the mitochondrial phospholipids of SSO-fed rats, the (22:4 n-6 + 22:5 n-6) to 18:2 n-6 ratio was decreased by HS, without modification of the proportion of 20:4 n-6. In the mitochondrial phospholipids of FO-fed rats, the amount of 20:5 n-3 tended to be higher (+56%). Cardiac functioning was modulated by the diets. Myocardial coronary flow was enhanced by HS in the SSO-fed rats, whereas it was decreased in the FO-fed animals. The rate constant k012 representing the activity of the adenylate kinase varied in the opposite direction, suggesting that decreased ADP concentrations could cause oxygen wasting through the opening of the permeability transition pore. The recovery of the pump function tended to be increased by n-3 PUFA feeding (+22%) and HS (+45%). However, the release of ascorbyl free radical during reperfusion was not significantly modified by the diets. Conversely, energy production was increased by ischemia/reperfusion in the SSO group, whereas it was not modified in the FO group. This supports greater ischemia/reperfusion-induced calcium accumulation in the SSO groups than in the FO groups. HS did not modify the mitochondrial energy metabolism during ischemia/reperfusion. Taken together, these data suggest that HS- and n-3 PUFA-induced decrease in 22:4 and 22:5 n-6 and increase in 20:5 n-3 favor the recovery of mechanical activity during post-ischemic reperfusion.

Measurement of free cholesterol, cholesteryl esters and cholesteryl linoleate hydroperoxide in copper-oxidised low density lipoprotein in healthy volunteers supplemented with a low dose of n-3 polyunsaturated fatty acids

A study was designed to evaluate the effect of supplementation with a low dose (0.9 g/d) of n-3 polyunsaturated fatty acids ( n-3 PUFA) in fish oil on the oxidative modification of low-density lipoprotein (LDL) in a group of healthy volunteers. Eight volunteers were randomly selected from a larger fish oil supplementation study, and were required to take either 0.9g n-3 PUFA as fish oil (FO group) or 0.9g olive oil [control oil (CO group)] for 16 weeks. Oxidative modification of LDL was assessed by measuring concentrations of free cholesterol (FC), cholesteryl esters (CE) and cholesteryl linoleate hydroperoxide (Ch18:2-OOH) in LDL following copper-induced lipid peroxidation for 0, 2, 3 and 4 h. The composition of LDL fatty acids over 4 h of copper-induced oxidation was also investigated. LDL eicosapentaenoic acid (C20:5 n-3) and docosahexaenoic acid (C22:6 n-3) compositions were significantly ( P < 0.05) higher in the FO compared with the CO group, following supplementation. Linoleic acid (C18:2 n-6), arachidonic acid (C20:4 n-6), C20:5 n-3 and C22:6 n-3 were significantly ( P < 0.05) oxidised in LDL following 4 h copper-oxidation. The proportions of palmitic acid (C16:0) ( P < 0.05), palmitoleic acid (C16:1) ( P < 0.05), stearic acid (C18:0), and oleic acid (C18:1) increased in the FO and CO groups, after 4 h of copper-oxidation. Concentrations of cholesteryl oleate (Ch18:1), cholesteryl linoleate (Ch18:2 n-6), cholesteryl arachidonate (Ch20:4 n-6) and cholesteryl docosahexanoate (Ch22:6 n-3) were significantly ( P < 0.05) reduced following copper-stimulated oxidation, in both groups. Ch18:2-OOH concentrations were significantly increased ( P < 0.05) following 3 h oxidation in both groups compared with 0 h copper-oxidation, but decreased after 4 h. There was no significant difference in concentrations of Ch18:2-OOH between the groups over the time-course of copper-mediated oxidation. The results of this study suggest that moderate dietary intakes of n-3 PUFA do not significantly influence the susceptibility of LDL to copper-induced oxidation in vitro.

Difference Spatial Distribution Function Analysis of Methanol and Ethanol Solutions.

Spatial distribution functions (SDFs), gOO(x, y, z) and gOH(x, y, z), which show the probability of atom-atom pair distribution between solute and solvent atoms, have been used to characterize the anisotropic structure of solutions in computer simulations. In this article, a new method for structural change analysis is proposed based on the difference spatial distribution function (DSDF) ΔgOO(x, y, z), and is applied to liquid water, and to infinitely dilute aqueous solutions of methoanol and ethanol. Based on the results of SDF in methanol and ehtanol solutions, the distribution of solvent water can be classified into three groups : hydrogen acceptor, hydrogen donor, and hydrophobic hydration regions. Regarding the effect fo hydrophobic groups, the DSDF (i.e. ΔgOO(x, y, z)=gOO(x, y, z)EtOH-gOO(x, y, z)MeOH) results indicated that the distribution volume of hydration water surrounding the alcohol molecule in the hydrogen acceptor region decreases with increasing steric bulk of the hydrophobic group. However, binding energy in this region is stabilized by an increase in the coordination number. Moreover, from the results of DSDF at 298 and 273 K (i.e. Δg<OO>(x, y, z, ΔT)=g<OO>(x, y, z, 298 K)-g<OO>(x, y, z, 273 K)), the distribution of hydration water spreads out over the region surrounding the hydrogen acceptor and donor. This result is reasonable since an increase in the fluctuation of the network structure results from a rise in temperature. The new DSDF method presented in this work should be widely applicable to structural change analysis of hydration structure for anisotropic solutions by computer siulation.

P53 tumor suppressor gene mutation and prognosis in 105 cases of bladder cancer--the relationship between mutation of the p53 gene with clinicopathological features and smoking

Alterations of the p53 tumor suppressor gene are the most common genetic change detected in human cancers. The incidence of p53 gene mutation in bladder tumor patients were studied and were compared with clinicopathological findings, smoking history and prognosis. Polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) was used for analysis from exon 4 to 9 of p53 gene in 105 cases of primary bladder tumors. p53 matations were detected in 38 or 105 patients (36.2%). Kaplan-Meier Survival curves fo groups wit or without p53 gene mutation show a statistically significant difference (p = 0.0098). The mutation of p53 gene in stages pT 1 pT 1. pT 2, pT 3, pT 4 was found in 2 of 12 (16.7%), 8 of 32 (25.0%), 10 of 25 (40.0%), 12 of 20 (60.0%), 6 of 16 (37.5%) and in grades I, II, III, was noted in 1 of 17 (5.9%), 16 of 49 (32.7%), 21 of 39 (53.8%) cases, respectively. Significant differences were found for groups with grade I and grade II-III (p = 0.0045) cancers and in cases of superficial (stage pTa-1) and muscle-invasive (pT 2-4) tumors (p = 0.0148). However, mutation of p53 was not related to either age or sex in 105 patients. Recurrence rates in stage pTa-1 superficial tumor group with or without p53 mutation showed a statistically significant difference (p = 0.0419). No statistically significant difference was noted between p53 mutation and habitual smoking as well as durations of smoking. p53 mutations occur more commonly in higher grades and later stages of bladder tumors. Our results suggest that the prognostic factor is linked to not only histological findings but also to the presence of p53 mutation. The mutations of the p53 gene may be involved in the late events of tumorigenesis and might be used as good molecular markers for prognosis in bladder tumor.