Adventitial cells do not contribute to neointimal mass after balloon angioplasty of the rat common carotid artery.

BACKGROUND: Previous studies suggest that the migration of adventitial cells into the neointima after balloon angioplasty might have an important role in vascular lesion formation. The current experiments were designed to study the migration of adventitial cells in response to mechanical injury of the rat carotid artery. METHODS AND RESULTS: Adventitial cells were stained in situ with PKH26, a fluorescent dye, after balloon angioplasty of the rat common carotid artery. Animals were killed at different time points, and tissue sections were examined under light and fluorescence microscopy. PKH26-labeled cells were detected exclusively in the adventitia. No labeled cells were present in the media or the neointima at any time point examined. A highly cellular neoadventitial layer composed of myofibroblasts exhibited an extensive proliferative response 3 days after injury over the entire adventitial circumference. CONCLUSIONS: Despite the prominent role that adventitial myofibroblasts seem to have in the postangioplasty remodeling process, they do not migrate to the medial or intimal layers in the rat carotid artery angioplasty model.

Transformation of nonvascular acellular tissue matrices into durable vascular conduits.

BACKGROUND: Prosthetic grafts commonly used for vascular reconstruction are limited to synthetics and cross-linked tissue grafts. Within these devices, graft infections are common, compliance mismatch is significant, and handling qualities are poor. Natural biological tissues that are unfixed have been shown to resist infections and be durable and compliant. A natural biological matrix that could be remodeled appropriately after implantation would be a desirable graft for vascular reconstruction. METHODS: SynerGraft tissue engineering strategies have been used to minimize antigenicity and produce stable unfixed vascular grafts from nonvascular bovine tissues. These grafts have replaced the abdominal aortas of 8 dogs that have been followed for up to 10 months. RESULTS: Early evaluation indicates rapid recellularization by recipient smooth muscle actin positive cells, which become arranged circumferentially, into the media. Arterioles were present in the adventitial areas and endothelial cells were seen to cover lumenal surfaces. After 10 months, grafts were patent and not aneurysmal. CONCLUSIONS: These data indicate that SynerGraft processing of animal tissues is capable of producing stable vascular conduits that exhibit long-term functionality in other species.

Recellularization of heart valve grafts by a process of adaptive remodeling.

The objective of this study was to investigate if function and durability of connective tissue grafts stems from in vivo revascularization and recellularization. Viability is important for durable valve performance, demonstrated by pulmonary autografts. A pattern of in vivo recellularization occurs in xenogeneic or allogeneic heart valves decellularized prior to implantation, dictated by the tissue matrix and functional biomechanics. Porcine or sheep heart valves were decellularized with the SynerGraft antigen reduction process (a common treatment process to remove all histologically demonstrable leaflet cells), and implanted as pulmonary (n = 11) or aortic valve (n = 9) replacements in sheep. Sheep allograft pulmonary valves (n = 4) were implanted as pulmonary valve replacements. Recellularization was evaluated histologically after 3, 4, 5, 6, and 11 months, with cell phenotypes identified using specific antibodies. SynerGraft heart valves were progressively recellularized beginning with an initial cellular infiltrate, and subsequent repopulation with mature interstitial cells. This process occurs in the conduit and then in the leaflet, and is associated with revascularization of the graft. Functional, fully developed fibrocytes, actively synthesizing type I procollagen (antibody probe) were present within 3 months. As the process matured cell density and distribution became similar to native valve leaflets with localization of smooth muscle actin positive cells at the ventricularis/spongiosa interface. After 11 months, leaflet explants had no detectable inflammatory cells, were as much as 80% repopulated, and had a distribution of smooth muscle actin positive cells similar to that of the natural leaflet. SynerGraft- treated heart valve implants are repopulated by a process typical of adaptive remodeling following implantation. This antigen reduction treatment is the first successful tissue engineering effort obtaining an implant with mature recipient cells capable of matrix protein synthesis. Normal early valve function and durability is maintained.

Simulating ozone detoxification in the leaf apoplast through the direct reaction with ascorbate.

This paper presents a mathematical model which enables the semi-quantification of ozone (O3) detoxification, based upon the direct reaction of the pollutant with ascorbate (ASC) located in the aqueous matrix associated with the cell wall (i.e. the apoplast). The model describes the uptake of ozone into the leaf and its direct reaction with ASC, taking into consideration the regeneration of dehydroascorbic acid in the cytosol, the rate of replenishment of cell wall ASC and the distribution of ASC between sub-cellular compartments based upon the permeability of biomembranes to the neutral species, ascorbic acid and the pH of various sub-cellular compartments. The importance of various physico-chemical characteristics (e.g. stomatal conductance, mesophyll cell wall thickness and tortuosity, chloroplast volume, apoplast pH, ASC:O3 reaction stoichiometry) in mediating the flux of ozone to the plasmalemma is analysed. Model simulations, supported by experimental observations, suggest that the ASC concentration in the leaf apoplast is high enough to scavenge a significant proportion of the O3 taken up into the leaf interior, under environmentally relevant conditions. However, there is considerable variation between taxa in the potential degree of protection afforded by apoplastic ASC, emphasizing the need for an improved understanding of the reaction chemistry of O3 in the cell wall.

Construction of phase cycles of minimum cycle length: MakeCycle

An algorithm for the generation of a phase cycle of minimum length for a pulse sequence is developed from the basic requirement that only specified coherence transfer pathways will be accumulated. The efficacy of the algorithm is shown by determining the phase cycles of minimum length for DQFCOSY, GHMBC, and INEPT pulse sequences. Copyright 2000 Academic Press.

Impacts of ozone on the growth and yield of field-grown winter wheat.

Seed of winter wheat (Triticum aestivum L. cv Riband) was sown on 29 August 1992 in eight field plots. Four plots were exposed to elevated ozone (O(3)) concentrations on 16 days between 29 August and 2 October 1992, for 6 h day(-1), and on 27 days between 29 March and 24 August 1993, for 7 h day(-1). Mean daily O(3) concentrations were approximately 30 and 80 nmol mol(-1) in ambient and fumigated plots, respectively. Plants were sampled on 5 November (1992), 14 January, 16 February, 1 April, 25 May, 23 June and 24 August (1993). No visible symptoms of O(3) damage or premature senescence were observed at any time over the course of the experiment. Exposure to elevated O(3) decreased the above ground biomass by reducing plant density and individual plant relative growth rate. However, there was no significant influence of the pollutant on the growth of the root relative to the shoot. Assessment of yield characteristics at the final harvest revealed an O(3)-induced decrease in the number of grains per ear, as a result of fewer grains per spikelet and an increase in the number of infertile florets per spikelet. No significant effects of the pollutant on the number of ears per plant, spikelets per ear, or 1000 grain weight were found. As a result of the combined effects on the number of grains per ear and the decrease in plant density and growth rate, O(3) exposure reduced grain and straw yields (tonnes ha(-1)) by 13 and 8%, respectively. However, no significant change in the partitioning of dry matter between the grain and the straw was observed in fumigated plots. The findings are discussed within the context of United Nation Economic Commission for Europe critical level guidelines for the protection of crop yields, in relation to their application to winter-sown crops.

Successful small diameter arterial grafting using cryopreserved allograft arteries.

Intimal hyperplasia (IH) limits the long-term success of veins as arterial grafts. IH occurs in veins partly as an adaptive process to arterial pressure conditions. The authors have previously reported early success with cryopreserved (CP) saphenous veins as aortocoronary bypass grafts, and they have hypothesized that CP arterial segments were already structurally adapted for arterial conditions. Six femoral arterial segments were harvested from three adult donor dogs, and cryopreserved. The segments were thawed and implanted into six recipient dogs, in end-to-end fashion, as interpositional grafts in the femoral artery. A similar length of native femoral artery was removed from the implant site and grafted in the contralateral femoral artery of the same animal to serve as native autograft-matched controls. Grafts were harvested bilaterally after 2 (n = 3) and 4 weeks (n = 3), perfusion fixed (80 mmHg, 15 min), and analyzed histologically. All grafts were patent at harvest, and flows distal to the grafted segments were not significantly different between grafts within an animal either at implant or subsequent harvest. Although CP arterial grafts still showed slight but significant dilation compared with native autograft, the dilation was much less than seen previously with either CP or native venous segments. No evidence of inflammation or IH was seen in CP arterial grafts. The absence of early IH or inflammation suggests that CP small diameter arteries may perform better than many currently available allograft tissues and synthetic prosthetics.

The effects of different ozone exposures on three contrasting populations of Plantago major.

Plantago major grows throughout Britain in a range of ozone climates. Because populations have been shown to differ in ozone resistance, the aim of the experiment was to compare the reaction of populations from contrasting ozone climates to different types of ozone exposure. Three populations were grown under controlled conditions in five different ozone treatments (including controls for 10 wk. Development, growth, stomatal conductance and seed production were recorded. Populations were from the south coast of England (Lullington), near a mountain summit (Great Dun Fell) and lowland Scotland (Bush). Ozone treatments were: charcoal and Purafil filtered air (CF); 35 nl l(-1) for 24 h every day: 70 nl l(-1) h for 7 h everyday: CF then three episodes each week of 70 nl l(-1) for 7 h; and 35 nl l(-1) continuously plus three 7 h episodes each week of 70 nl l(-1) . The different ozone treatments resulted in different responses in each population. Ozone promoted senescence in the Great Dun Fell population but not in the others; it reduced root growth more in the Lullington population than in the others but those from Lullington and Great Dun Fell maintained seed production to a much greater extent than the Bush population. The reproductive effort (number of seeds g(-1) of vegetative weight) actually increased in ozone in the Lullington and Great Dun Fell populations. It is suggested that this might he a general stress response rather than being specifically related to ozone. Effects on stomatal conductance were similar to those previously reported and the converse of effects on seed production. The relative responses of the populations varied according to the ozone treatment. Continuous exposure to 35 n1 l(-l) reduced leaf size only in the Great Dun Fell population, but seed output was reduced in the Bush population. In some cases, giving 3-d episodes of 70 n1 l(-1) had a greater effect than giving the dose every day but the effects varied with the population. This greater effect was considered to be a result of the time it takes for a plant to develop maximum anti-oxidant defence, which is lost when the ozone decreases after the episode. A plant exposed to episodes might have to re-induce defence with each exposure. Although it is reported frequently that ozone favours allocation of resources to the shoot over the root, it is concluded that this is an over-simplification of the response. Even within a species there is a complex suite of responses that varies with the population and with ozone exposure. Describing a population as resistant or sensitive is also an over-simplification.

The use of an unenclosed field fumigation system to determine the effects of elevated ozone on a grass-clover mixture.

A mixture of Trifolium repens L. (var. Grasslands Huia) and Lolium perenne L. (var. Melle) was established in an area with low ambient ozone concentrations and exposed to episodes of ozone using a simple, unenclosed field fumigation system. Control plots were exposed to ambient air. There were two cutting frequencies to simulate grazing and silage production respectively. This paper reports on the performance of the system and the effects on yield and species composition in the first year of fumigation. The system operated satisfactorily when the wind speed was between 1 and 5 m s-1 . When the wind dropped suddenly below 1 m s-1 there were short periods with high concentrations but these events were rare and did not last more than a few minutes. Above 5 m s-1 there was lateral downwind spillage of ozone and at higher wind speeds it was not possible to maintain the target concentration of 50-70 nl 1-1 . The plots were fumigated only when the weather was suitable for ozone formation and when the wind speed was stable. Because the weather was poor, fumigation was restricted to 22 d between July and September. The mean 4-7 h concentrations were 50-70 nl l-1 . Seven-hour mean ambient concentrations over the same period were 10-28 nl 1-1 . There were significant effects of ozone on yield and herbage composition. The effects varied between harvests, probably because of differences in the timing of cutting relative to the episodes, and to the differences in phenology between the species. Most of the loss of yield was due to effects on Trifolium, which was much more sensitive than Lolium. There was a persistent effect of ozone on the stolon density of Trifolium which lasted into the following year. It is suggested that this arose because the clover replaced damaged leaves very quickly, maintaining the canopy at the expense of the stolons. Persistent effects on stolons might lead to poorer winter survival and lower nitrogen fixation. Gaps in the sward as a result of loss of Trifolium might allow rapid weed invasion. There was no interaction between cutting frequency and ozone, which may have been due to the fact that the poor summer restricted the number of cuts and therefore decreased the difference between the two cutting frequencies.

Characterization of thrombin receptor expression during vascular lesion formation.

Blood vessels respond to injury by initiating cell proliferation and migration that result in vascular lesion formation. To determine the roles of thrombin and the thrombin receptor in this process, we characterized thrombin receptor expression in normal and injured arteries, thrombin receptor-mediated smooth muscle cell mitogenesis, and the regulation of thrombin receptor mRNA expression in vitro. Thrombin receptor mRNA was not detected in normal rat or baboon arteries by in situ hybridization. Immunohistochemistry using an antithrombin receptor antibody (TR-R9), directed against the thrombin cleavage site of the rat aortic smooth muscle cell thrombin receptor, revealed low-level staining for thrombin receptor protein in endothelial cells and smooth muscle cells of normal arteries. In contrast, balloon catheter injury increased thrombin mRNA expression in medial smooth muscle cells within 6 hours. This increased thrombin receptor expression continued within the media and in neointimal cells throughout vascular lesion formation, predominantly in areas of active cell proliferation. In vitro, alpha-thrombin stimulates rat aortic smooth muscle cell proliferation in a concentration-dependent manner. That thrombin receptor activation is required for the mitogenic response was confirmed by demonstrating that the polyclonal antibody TR-R9 inhibits thrombin-induced cell proliferation. Thrombin receptor mRNA synthesis was induced by both basic fibroblast growth factor (maximal stimulation of 1.8-fold at 1 hour) and platelet-derived growth factor (maximal stimulation of 2.4-fold at 8 and 24 hours) in quiesced cultured rat aortic smooth muscle cells. In summary, upregulation of smooth muscle cell thrombin receptor expression occurs very early after vascular injury and continues throughout neointimal development.(ABSTRACT TRUNCATED AT 250 WORDS)

Agonist-induced phosphorylation of the vascular type 1 angiotensin II receptor.

Agonist-induced receptor phosphorylation plays a role in transmembrane signal transduction systems. Although the cDNA for the rat vascular type 1 angiotensin II receptor (AT1AR) encodes a G protein-coupled receptor with several potential phosphorylation sites for serine/threonine and tyrosine kinases, little is known about the phosphorylation of this receptor. The aim of this study was to determine the effects of angiotensin II (Ang II) on phosphorylation of the AT1AR in rat aortic vascular smooth muscle cells. Using [32P]orthophosphate-labeled cells, immunoprecipitates with anti-AT1AR antibody revealed a labeled band of molecular weight 52 kD, corresponding to the Ang II receptor. Ang II induced a rapid and significant increase in phosphorylation of the Ang II receptor, with a peak at 20 minutes. Phosphoamino acid analysis showed that the major phosphoamino acid is serine, in both the basal and Ang II-stimulated states. Constitutive and agonist-stimulated tyrosine phosphorylation is also observed to a lesser extent. Immunoblotting of anti-phosphotyrosine immunoprecipitates with anti-AT1AR antibody showed that Ang II caused a delayed tyrosine phosphorylation of the receptor with a peak at 20 minutes in a dose-dependent manner. Forskolin increased total phosphorylation of AT1AR but had no effect on tyrosine phosphorylation. Neither phorbol 12-myristate-13-acetate nor ionomycin altered receptor phosphorylation. These findings suggest that Ang II induces the phosphorylation of its own G protein-coupled receptor through both serine and tyrosine kinases and raise the possibility that phosphorylation of the AT1AR is an important regulator of receptor function.

Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells.

The signaling pathways involved in the long-term metabolic effects of angiotensin II (Ang II) in vascular smooth muscle cells are incompletely understood but include the generation of molecules likely to affect oxidase activity. We examined the ability of Ang II to stimulate superoxide anion formation and investigated the identity of the oxidases responsible for its production. Treatment of vascular smooth muscle cells with Ang II for 4 to 6 hours caused a 2.7 +/- 0.4-fold increase in intracellular superoxide anion formation as detected by lucigenin assay. This superoxide appeared to result from activation of both the NADPH and NADH oxidases. NADPH oxidase activity increased from 3.23 +/- 0.61 to 11.80 +/- 1.72 nmol O2-/min per milligram protein after 4 hours of Ang II, whereas NADH oxidase activity increased from 16.76 +/- 2.13 to 45.00 +/- 4.57 nmol O2-/min per milligram protein. The NADPH oxidase activity was stimulated by exogenous phosphatidic and arachidonic acids and was partially inhibited by the specific inhibitor diphenylene iodinium. NADH oxidase activity was increased by arachidonic and linoleic acids, was insensitive to exogenous phosphatidic acid, and was inhibited by high concentrations of quinacrine. Both of these oxidases appear to reside in the plasma membrane, on the basis of migration of the activity after cellular fractionation and their apparent insensitivity to the mitochondrial poison KCN. These observations suggest that Ang II specifically activates enzyme systems that promote superoxide generation and raise the possibility that these pathways function as second messengers for long-term responses, such as hypertrophy or hyperplasia.

Angiotensin-II-and endothelin-induced protein phosphorylation in cultured vascular smooth muscle cells.

In cultured vascular smooth muscle cells, angiotensin II and endothelin stimulate a variety of intracellular signals, including generation of inositol trisphosphate and diacylglycerol, mobilization of intracellular calcium, and activation of protein kinase C. These latter two events have been shown to mediate the phosphorylation of numerous proteins, but these substrates and the specific pathways mediating their phosphorylation have not been identified in vascular smooth muscle. Angiotensin II (100 nM, 10 min) induced a characteristic pattern of protein phosphorylation, which included the phosphorylation of many proteins, ranging in molecular mass from 20 to 76 kD. Three of these proteins have been identified as vimentin (M(r) 57,000), a specific protein kinase C substrate (M(r) 76,000) and the myosin light chain (M(r) 20,000). The 76-kD protein was one of the most highly phosphorylated proteins after agonist treatment. Endothelin-1 produced an identical pattern of phosphorylation. Five of these substrates were also phosphorylated by phorbol-12-myristate-13-acetate, and 5 were also phosphorylated after treatment with ionomycin. In general, the protein-kinase-C-dependent phosphorylations were sustained, while those mediated by calcium were rapid. Since these experiments were performed in cultured, phenotypically modulated cells stimulated with agents that promote cellular hypertrophy or hyperplasia, this pattern of phosphorylation may be representative of that seen during the growth response.

Physiological effects of ozone on cultivars of watermelon (Citrullus lanatus) and muskmelon (Cucumis melo) widely grown in Spain.

Two cultivars of watermelon (Citrullus lanatus) and muskmelon (Cucumis melo), which are widely grown in Spain, were exposed to ozone (70 nl litre(-1), 6 h d(-1)) for 21 days. Ozone sensitivity was assessed by recording the extent of visible injury, changes in fast-fluorescence kinetics, the relative-growth rate (R) of root (RR) and shoot (RS), and effects on the number of flowers produced per plant. Leaf gas exchange was measured in order to provide some indication of the factors underlying differential response to ozone. After 9-10 days of fumigation, all the cultivars developed typical visible symptoms of zone injury on the older leaves. However, significant (P < 0.05) changes in fast-fluorescence kinetics were detected prior to the development of visible foliar injury, indicating that detectable effects of ozone on primary photochemical processes supersede the appearance of visible symptoms. In both muskmelon and watermelon, there was a marked reduction in the rate of CO(2) assimilation as a result of exposure to ozone, and this was accompanied by a parallel decrease in stomatal conductance. Mean plant-relative-growth rate (R) was markedly (P < 0.01) suppressed by ozone in the two cultivars of watermelon, but there were no significant effects on R in muskmelon. Ozone reduced root growth relative to the shoot in three out of four cultivars-an effect that may be of considerable ecological significance. Moreover, exposure to ozone reduced flower production in both muskmelon and watermelon, which indicated effects on yield. There was no correlation between a variety of methods used to assess ozone sensitivity and visible injury, and reasons for this are discussed. This observation draws clear attention to the dangers in ranking plants for ozone sensitivity purely on the basis of visible symptoms. It is concluded from this study that ozone-insensitive genotypes should be identified and considered for planting in the major areas of melon production concentrated on the Mediterranean coast of Spain.

The effect of short chain fatty acid supplementation on membrane electrolyte transport and blood pressure.

Eleven normotensive subjects with no family history of essential hypertension took part in a double-blind randomized placebo-controlled crossover study to examine the effects of supplementing a normal omnivore diet with miglyol. This resulted in a fall in diastolic blood pressure in both the supine and standing positions, achieving statistical significance for the standing diastolic pressures, following miglyol treatment. Miglyol is rich in caprylic (8:0) and capric acids (10:0), both short chain saturated fatty acids, and supplementation with this produced a significant fall in erythrocyte membrane oleic and linoleic acid (P less than 0.01 compared to placebo for each fatty acid), as well as a fall in the saturated fat palmitic acid (16:0) (P less than 0.01). These changes were not associated with any alterations in total erythrocyte sodium influx, bumetanide sensitive influx or sodium red cell intracellular or potassium content. In addition, body weight and urinary excretion of sodium and potassium did not change. These data indicate that this dietetic manipulation with an oil rich in short chain saturated fatty acids lowers diastolic blood pressure but not as a result of changes in membrane sodium handling. It is possible that the short chain fats displace the longer carbon chain fatty acids which are metabolically important to cellular integrity and it is in this way that blood pressure falls.

Resistance artery phosphoinositide metabolism in genetic hypertension.

Precapillary resistance arteries from spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive rats (WKY) were found to contain three inositol lipids and to produce five inositol phosphate peaks. These were assessed by a highly sensitive procedure which involved the separation of radiolabelled inositol-containing components by anion-exchange high-performance liquid chromatography. Basal levels of radiolabelled inositol lipids were found to be significantly increased in SHR at 5 weeks of age, and also increased at 12 weeks, although this was only statistically significant for glycerophosphoinositol. At 5 weeks of age, exposure to a maximal concentration of noradrenaline brought about a significant increase in lipid radioactivity in WKY and in glycerophosphoinositol and glycerophosphoinositol 4-phosphate in SHR. The levels of these lipids remained significantly raised in the SHR at this time. At 12 weeks of age, exposure to noradrenaline produced reductions in radioactivity associated with inositol lipids. These new levels were elevated in SHR and significant for glycerophosphoinositol 4-phosphate. Basal levels of four inositol phosphates were not different between the two rat strains at 5 weeks of age. Inositol 1,3,4-trisphosphate was only present at minute levels and could not be measured. At 12 weeks of age, basal radiolabelling of inositol phosphates was not different between the two strains. The application of noradrenaline caused age-dependent changes in arterial inositol-containing compounds in both strains, which resulted in increased levels of inositol 1,4,5-trisphosphate in young SHR and also in adult rats in which the blood pressure level had become established.(ABSTRACT TRUNCATED AT 250 WORDS)

Agonist-induced production of 1,2-diacylglycerol and phosphatidic acid in intact resistance arteries. Evidence that accumulation of diacylglycerol is not a prerequisite for contraction.

The production of total amounts of 1,2-diacylglycerol as well as those specifically derived from inositol lipid hydrolysis was studied in intact rat resistance arteries stimulated with either noradrenaline, vasopressin, or angiotensin II at 20 s when the onset of contraction would be nearing its maximum, and at 5 min during the sustained phase of contraction. Total amounts of 1,2-diacylglycerol were not altered by any agonist at 20 s, or at 5 min. However, arachidonate-containing species of 1,2-diacylglycerol were differentially influenced being increased at 5 min by noradrenaline, and decreased at 20 s and 5 min by vasopressin. Only angiotensin II produced substantial increases in this class of 1,2-diacylglycerol at both time points. In order to investigate the fate of this second messenger total and inositol lipid derived phosphatidic acids were then measured at both 20 s and 5 min. Noradrenaline induced a rise in both total and arachidonate-containing phosphatidic acid at both times as did vasopressin. Only small increases were induced by angiotensin II at 20 s. These data demonstrate that the accumulation of 1,2-diacylglycerol generated from inositol lipid breakdown is only observed with activation by angiotensin II. Other agonists produced phosphatidic acids with time and the rate of generation of these lipids is agonist-specific. Thus phosphatidic acid may play a more prominent role during the sustained phase of contraction than previously anticipated.

Linoleic acid supplementation, membrane lipids and leucocyte sodium transport in normotensive humans.

Ten normotensive subjects had their omnivore diet supplemented with increasing doses of linoleic acid in the form of safflower seed oil in order to examine the effects of this polyunsaturated fat upon leucocyte sodium transport. Increasing the dose ingested to the limits of tolerance produced a significant decrease in ouabain resistant sodium efflux (P less than 0.05) but no significant change in total or ouabain-sensitive sodium extrusion. Intraleucocytic sodium content was correlated with erythrocyte membrane oleic acid % (r = 0.368, P less than 0.05); leucocyte ouabain resistant flux was correlated with oleic acid % (r = 0.453, P less than 0.01) and linoleic acid % (r = -0.319, P less than 0.05). No such associations were observed with ouabain sensitive sodium extrusion. No changes in body weight or urinary electrolyte excretion were observed. It is concluded that, at physiological concentrations, membrane linoleic acid content influences transmembrane sodium fluxes but not through modulation of sodium pump activity. Furthermore, the beneficial effect of this dietary manoeuvre, observed previously with small increments of safflower seed oil, was not seen in this experiment so the hypotensive activity of this manipulation must be regarded as limited.

The effect of dietary linoleic acid on blood pressure and erythrocyte sodium transport.

The influence of an increase in the polyunsaturated fat linoleic acid on blood pressure and erythrocyte membrane sodium transport was investigated in normotensive first degree relatives of hypertensive patients and controls by the double blind administration of safflower oil or paraffin oil (placebo) capsules for four weeks separated by a four week washout period. Systolic blood pressure fell in the controls with linoleic acid supplementation but there was no significant change in total sodium efflux rate constant. When the pattern of response was compared the changes in supine systolic blood pressure, plasma renin activity, total and ouabain-sensitive sodium efflux rate constant were significantly different in the controls compared to the relatives. These results show that dietary linoleic acid supplementation may have effects on ionic fluxes across cell membranes and cause a modest fall in blood pressure. In addition, since the response to the change in fat intake was different in the relatives and the controls, this provides further evidence of differences in the physicochemical structure of the plasma membrane in hypertensive subjects and their offspring.