The interindividual variation in ApoE plasma concentration is considerable, mainly determined by apoE genotype and sex. However, a large amount of variability remains unexplained by these factors. We have evaluated whether the quantity and quality of dietary fat interacts with the apoE genotype and sex modifying ApoE plasma levels in young healthy subjects. Eighty-four volunteers (sixty-six apoE3/3, eight apoE4/3 and ten apoE3/2) were subjected to three dietary periods, each lasting 4 weeks. The first was a SFA-enriched diet (38 % fat and 20 % SFA), which was followed by a carbohydrate (CHO)-rich diet (30 % fat, < 10 % SFA and 55 % carbohydrate) or a MUFA-rich diet (38 % fat and 22 % MUFA) following a randomised crossover design. apoE2 carriers have the highest ApoE levels, whereas apoE4 individuals show the lowest concentration after the SFA, CHO and MUFA diets. Women had significantly higher ApoE concentration than men only after the consumption of the SFA diet. The SFA diet increased the ApoE plasma concentration when compared with the CHO- and MUFA-rich diets in women, but not in men. In women, but not in men, the shift from the SFA- to CHO- or MUFA-rich diets significantly decreased the ApoE concentration in apoE3/2 and apoE3/3 subjects, whereas no differences were observed in women with the apoE4/3 genotype. Sex and apoE genotype determine ApoE plasma levels; however, this effect is dependent on dietary fat.

Bowman–Birk inhibitor (BBI) from soyabeans is a naturally occurring protease inhibitor with potential anti-inflammatory and chemopreventive properties within the gastrointestinal tract (GIT). In a previous paper, we reported that significant amounts of BBI-related proteins reach the terminal ileum functionally and biologically active. We have now investigated: (a) if soyabean BBI is biotransformed by faecal microbiota which would reduce its potential colorectal chemopreventive properties and (b) the potential influence of this protease inhibitor on the modulation of faecal microbiota. In vitro incubation studies of native soyabean BBI at a physiological level (93 μm) with mixed faecal samples of pigs for 24 h at 37°C demonstrated that BBI remains active and its intrinsic trypsin and chymotrypsin inhibitory activities were not significantly influenced by the enzymic or metabolic activity of faecal microbiota. Soyabean BBI did not affect the growth of the different bacterial groups studied (lactobacilli, bifidobacteria, bacteroides, coliforms, enterobacteria, clostridia and total anaerobes). It was concluded that protease inhibitory activities, intrinsically linked to the chemopreventive properties of soyabean BBI, were largely unaffected by faecal microbiota in vitro. BBI retains significance, therefore, as a bioactive compound in the human GIT.

The effect of amylose content on digestibility of starch in the small intestine and on the concentration of plasma lipid were studied in ileorectostomized rats and in ovariectomized rats, respectively. Seven kinds of starch with different amylose content (0, 27, 54, 62, 76, 79, 86 %) were used as test starch, which contained 0·4, 5·6, 37·1, 40·2, 45·6, 36·9 and 36·1 % resistant starch (RS), respectively. Rats were fed one of test diets containing 30 % test starch with different amylose content for 14 d in ileorectostomized and for 21 d in ovariectomized rats. Food intake was not significantly different among the groups. In ileorectostomized rats, the small intestinal starch digestibility decreased with increasing intakes of amylose and RS. In ovariectomized rats, body weight gain was lower on the higher amylose maize starch diets. The concentrations of plasma TAG and cholesterol decreased with increasing intake of RS. The concentrations of liver total lipids and TAG decreased with increasing intake of RS, but that of liver cholesterol did not. There was significant positive correlation between the level of sterol regulatory element-binding protein-1c mRNA and concentration of liver TAG. Total SCFA amount in the caecum increased logarithmically with increasing dry weight of caecal contents. The amount of bile acids in the small intestinal content and the excretions of bile acids and neutral steroids in faeces increased with increasing RS intake. These results show that starch rich in RS is more effective in preventing ovarian hormone deficiency-induced hyperlipidaemia.

Contrasting data are published on the effects of high salt intake (between 300 and 660 mmol/d) on Na balance and fluid retention. In some studies high levels of NaCl intake (400, 440, 550 and 660 mmol/d) led to positive Na balances without fluid retention. To test the relevance of different baseline NaCl intake levels on changes in metabolic water, Na, K, chloride and acid–base balance, a 28 d clinical trial (‘Salty Life 6’) was carried out in a metabolic ward. Nine healthy male volunteers (aged 25·7 (sd 3·1) years; body mass (BM) 71·4 (sd 4·0) kg) participated in the present study. Four consecutive levels of NaCl intake: low (6 d, 0·7 mmol NaCl/kg BM per d), average normal (6 d, 2·8 mmol NaCl/kg BM per d), high (10 d, 7·7 mmol NaCl/kg BM per d), and low again (6 d, 0·7 mmol NaCl/kg BM per d) were tested. Urine osmolality, extracellular volume (ECV) and plasma volume (PV), cumulative metabolic Na, K, chloride and fluid balances, mRNA expression of two glycosaminoglycan (GAG) polymerisation genes, capillary blood pH, bicarbonate and base excess were measured. During average normal NaCl intake, 193 (sem 19) mmol Na were retained and ECV (+2·02 (sem 0·31) litres; P < 0·001) and PV (+0·57 (sem 0·13) litres; P < 0·001) increased. During high NaCl intake, 244 (sem 77) mmol Na were retained but ECV did not increase (ECV − 0·54 (sem 0·30) litres, P = 0·089; PV +0·27 (sem 0·25) litres, P = 0·283). mRNA expression of GAG polymerisation genes increased with rise in NaCl intake, while pH (P < 0·01) and bicarbonate (P < 0·001) levels decreased. We conclude that a high NaCl intake may increase GAG synthesis; this might play a role in osmotically inactive Na retention in humans.

Animals may have target levels for lipid and protein stores which they try to maintain by feedback mechanisms. Thus, variation in initial body composition may be related to subsequent feed utilisation, for animals to maintain body composition in homeostasis. We assessed whether such relationships are genetically determined within a farmed population of European whitefish (Coregonus lavaretus) grown either on fishmeal or soyabean-meal diets. Soyabean meal is an increasingly-used ingredient in aquaculture feeds. Fish from thirty-five paternal families were analysed for initial body lipid and protein content, and for subsequent daily weight gain, daily feed intake, feed efficiency and their lipid and protein components. The results showed that none of the correlations of initial body lipid percentage with subsequent growth and feed utilisation were statistically significant. In contrast, low initial protein percentage was related to increased subsequent weight gain, protein gain and protein retention efficiency. This led to reversed ranking of families during growth for body protein percentage. Thus, mechanisms maintaining stable body lipid percentage across the population were weak, whereas the mechanisms stabilising body protein percentage were strong and successful. This explains the observations that cascades of lipid deposition occur during fish growth, leading to high amounts of phenotypic and genetic variation for percentage body lipid. In contrast, protein percentage remains phenotypically and genetically more invariable, reducing the potential for selective breeding. The soyabean-meal diet, in turn, induced only weak genotype × diet interactions, aiding in the genetic improvement of farmed fish to adapt to future feeds.

Dietary patterns have been related to health outcomes and morbi-mortality. Mediterranean diet indexes are correlated with adequate nutrient intake. The objective of the present study was to analyse the adequacy of nutrient intake of a posteriori defined Mediterranean (MDP) and Western (WDP) diet patterns in the Seguimiento Universidad de Navarra (SUN) cohort. A sample of 17 197 subjects participated in the study. Participants completed a 136-item validated semi-quantitative FFQ. Principal component analysis was used to define dietary patterns. Individuals were classified according to quintiles of adherence based on dietary pattern scores. Non-dietary variables, such as smoking and physical activity habits, were also taken into account. The probability approach was used to assess nutrient intake adequacy of certain vitamins (vitamins B12, B6, B3, B2, B1, A, C, D and E) and minerals (Na, Zn, iodine, Se, folic acid, P, Mg, K, Fe and Ca). Logistic regression analysis was used to assess the adequacy of nutrient intake according to adherence to dietary patterns. WDP and MDP were defined. A higher quintile of adherence to an MDP was associated to a lower prevalence of inadequacy for the intake of Zn, iodine, vitamin E, Mg, Fe, vitamin B1, vitamin A, Se, vitamin C and folic acid. The adjusted OR for not reaching at least six (or at least ten) nutrient recommendations were 0·09 (95 % CI: 0·07, 0·11) (and 0·02 (95 % CI: 0·00, 0·16)) for the upper quintile of MDP and 4·4 (95 % CI: 3·6, 5·5) and 2·5 (95 % CI: 1·1, 5·4) for the WDP. The MDP was associated to a better profile of nutrient intake.

Increasing evidence suggests a role for prenatal environment in the onset of cardiovascular and metabolic disease in later life. In the rat, undernutrition in utero and a postnatal high-fat diet gives rise to a phenotype similar to the metabolic syndrome. As endothelial dysfunction is a feature of both CVD and the metabolic syndrome we investigated the impact of maternal undernutrition and/or postnatal high-fat on endothelial function. Virgin Wistar rats were mated and randomly assigned to groups to receive food either ad libitum (control) or at 30 % of ad libitum intake throughout gestation. At postnatal day 250, a cohort from each group was challenged with a high-fat diet (D12451, 45 % energy from fat; Research Diets, Inc., New Brunswick, NJ, USA) for the remainder of the study. At 1 year of age, small mesenteric arteries were dissected and mounted on a wire myograph and responses to phenylephrine, endothelin, acetylcholine, leptin and sodium nitroprusside assessed. Vasoconstriction to endothelin was significantly enhanced in all groups compared with controls (-log effective concentration equal to 50 % of the maximal response (pEC50); P < 0·001). Endothelium-dependent vasodilatation to acetylcholine was significantly blunted in all groups compared with controls (% maximum response; P < 0·01), while dilatation to leptin and sodium nitroprusside was similar in all groups. These data demonstrate that both maternal undernutrition and postnatal high fat lead to vascular alterations and suggest that maternal undernutrition alone is at least as detrimental to offspring endothelial function as a long-term exposure to a high-fat diet in the offspring.

The aim of this work was to compare the efficacy of two strategies designed to improve folate status: increasing the intake of vegetables, and the consumption of a folic acid-fortified food. Residents (126) from three old people's homes in the Madrid region (Spain) were studied. To each centre a dietary intervention was assigned to be followed for 6 months: (1) the consumption of margarine fortified with 200 μg folic acid/10 g portion (centre M), (2) increasing the consumption of vegetables to three servings per day (centre V), (3) control (centre C). At the beginning and end of the intervention period the subjects' intakes, serum and erythrocyte concentrations of folate were measured. The use of fortified margarine (centre M) led to a significant increase in folate intake (260·9 μg/d), serum concentration (10·3 (sd 8·3) nmol/l) and erythrocyte concentration (638·4 nmol/l). At centre V the increase in total vegetable intake achieved was very poor; these foods met with very poor acceptance, although the intake of certain vegetables particularly rich in folate improved. Therefore, the intake of this vitamin increased a little (26·7 (sd 33·0) μg/d); erythrocyte folate concentration also increased somewhat (460·5 nmol/l), althought less than centre M. The daily consumption of margarine fortified with folic acid was the more effective strategy for improving the folate status of the study subjects.

Dietary cholesterol may influence Alzheimer's disease risk, because it regulates the synthesis of amyloid-β. It was recently demonstrated in enterocytes of wild-type mice that intracellular amyloid-β expression is enhanced in response to a high-fat diet made up of SFA and cholesterol. Intestinally derived amyloid-β may be associated with postprandial lipoproteins in response to dietary fats and could be a key regulator in chylomicron metabolism. The present study was designed to investigate the role of cholesterol in modulating amyloid-β abundance in enterocytes. Wild-type mice were fed a low-fat diet supplemented with 2 % (w/w) cholesterol. The effects of cholesterol absorption inhibition and cholesterol biosynthesis inhibition utilising ezetimibe and atorvastatin, respectively, were also studied. Quantitative immunohistochemistry was utilised to determine enterocytic amyloid-β homeostasis. We found that enterocytic amyloid-β concentration was significantly attenuated in mice fed the 2 % (w/w) cholesterol diet. However, blocking cholesterol absorption reversed the cholesterol-feeding effect. Consistent with a suppressive effect of cholesterol on enterocytic amyloid-β abundance, atorvastatin, an inhibitor of cholesterol biosynthesis, enhanced amyloid-β. However, providing exogenous cholesterol abolished the atorvastatin-induced effect. In contrast to the suppression of enterocytic amyloid-β by dietary cholesterol, mice fed a diet enriched in SFA had markedly greater abundance. Collectively, the findings suggest that exogenous and endogenous cholesterol reduce amyloid-β concentration in enterocytes by suppressing production, or enhancing secretion associated with postprandial lipoproteins. Intestinally derived amyloid-β will contribute to the pool of plasma protein and may influence cerebral amyloid homeostasis by altering the bi-directional transfer across the blood–brain barrier.

Inflammatory bowel disease (IBD) is often associated with malnutrition. The aim of this study was to compare the body composition of outpatients with IBD during remission and active phase. In order to evaluate disease activity we used Crohn's Disease Activity Index for Crohn's disease (CD) patients and Lichtiger's Index for ulcerative colitis (UC) patients. All patients underwent the analysis of BMI, arm muscle area (AMA) and triceps plus subscapula skinfold thickness (TST+SST) to identify total, muscle and fat mass, respectively. In total 102 patients were evaluated (CD, n 50; UC, n 52) and the majority was young women. Malnutrition according to BMI was found in 14·0 % of patients with CD and 5·7 % of UC patients. Muscle mass depletion was detected in more than half of the CD and UC patients. The BMI, TST+SST and AMA values were lower in the active phase only in CD patients (P < 0·05). Fat mass depletion was associated with active phase in both CD and UC patients. Body composition parameters obtained using BMI, TST+SST and AMA were not correlated with the presence of fistula in CD patients (P>0·05). In conclusion, patients without signs of malnutrition had fat mass depletion especially in the active phase and muscle mass depletion occurred both in CD and UC patients.

EURopean micronutrient RECommendations Aligned is a network of excellence funded by the European commission, and established to address the problem of differences between countries in micronutrient recommendations as well as to understand how nutritional information including requirements and adequacy of intake is processed among different population groups. The aims of the present paper were to review the methods used for the adequacy assessment of the intake of six micronutrients of public health concern (vitamin A, folate, vitamin B12, Fe, Zn and iodine) in non-European and European nutrition surveys carried out on the apparently healthy population and to compare in particular the adequacy across surveys for folate intake. A systematic literature review was conducted to identify nutrition surveys that assessed micronutrient intake adequacy. The search yielded 9049 records, out of which 337 were eligible for the selected micronutrients. The majority (83·9 %) of the European surveys compared the adequacy of the nutrient intake against the Recommended Dietary Allowances (RDA); only a few surveys (8·0 %) used the estimated average requirement cut-point method, while none of them used the probability approach. The comparison of folate inadequacy across eight countries revealed that about 25 % of the adult female population had inadequate intakes when judged against the different recommendations used by the respective investigators, but nearly 75 % had inadequate intakes when compared against the estimated average requirement cut-point value of 320 μg/d. The present review showed that different methods were applied across Europe to estimate the adequacy of micronutrient intake, which led to different prevalence estimates of micronutrient inadequacy.

Four replicated groups of sea bass (Dicentrarchus labrax) larvae were fed diets containing an extra-high level of highly unsaturated fatty acids (HUFA) (XH; 3·7 % EPA+DHA), a high level of HUFA (HH; 1·7 %), a low level of HUFA (LH; 0·7 %) or an extra-low level of HUFA (XLH; 0·5 %) from day 6 to day 45 (experiment 1; XH1, HH1, LH1, XLH1). After a subsequent 1-month period feeding a commercial diet (2·7 % EPA+DHA), the capacity of the four initial groups to adapt to an n-3 HUFA-restricted diet (0·3 % EPA+DHA; R-groups: XH2R, HH2R, LH2R, XLH2R) was tested for 35 d. Larval dietary treatments had no effect on larval and juvenile survival rates. The wet weight of day 45 larvae was higher in XH1 and HH1 (P < 0·001), but the R-juvenile mass gains were similar in all treatments. Δ-6-desaturase (Δ6D) mRNA level was higher in LH1 and XLH1 at day 45 (P < 0·001), and higher in LH2R and XLH2R, with a significant increase at day 118.Concomitantly, PPARα and PPARβ mRNA levels were higher in XLH1 at day 45, and PPARβ and γ mRNA levels were higher in XLH2R at day 118, suggesting possible involvement of PPAR in stimulation of Δ6D expression, when drastic dietary larval conditioning occurred. The low DHA content in the polar lipids (PL) of LH1 and XLH1 revealed an n-3-HUFA deficiency in these groups. Larval conditioning did not affect DHA content in the PL of R-juveniles. The present study showed (i) a persistent Δ6D mRNA enhancement in juveniles pre-conditioned with an n-3 HUFA-deficient larval diet, over the 1-month intermediate period, and (ii) brought new findings suggesting the involvement of PPAR in the Δ6D mRNA level stimulation. However, such nutritional conditioning had no significant effect on juvenile growth and lipid composition.

This study was performed to investigate whether dietary lysine concentration influences the carnitine status of pigs. Therefore, an experiment with twenty young pigs with an average body weight of 21 kg was performed which were fed either a control diet (9·7 g lysine/kg) or a diet with a moderate excess of lysine (16·8 g lysine/kg). Concentrations of all the other amino acids did not differ between the diets. Pigs fed the high-lysine diet had lower concentrations of free and total carnitine in plasma, liver, kidney and skeletal muscle than control pigs (P < 0·05). Pigs fed the high-lysine diet moreover had an increased concentration of trimethyllysine (TML), a reduced mRNA abundance of TML dioxygenase and reduced concentrations of γ-butyrobetaine (BB) in muscle, indicating that the conversion of TML into BB in muscle was impaired. Concentrations of BB, the metabolic precursor of carnitine, in plasma, liver and kidney were also reduced in pigs fed the high-lysine diet while the activity of BB dioxygenase in kidney was not different and that in liver was even increased compared to control pigs (P < 0·05). In conclusion, this study shows that a moderate dietary excess of lysine lowers plasma and tissue carnitine concentrations in pigs. Reduced concentrations of BB in liver and kidney suggest that the depressed carnitine status was likely caused by a decreased rate of carnitine synthesis due to a diminished availability of carnitine precursor, probably mainly as a result of an impaired BB formation in muscle.

The effects of ascorbic acid (AA), phytate and tannic acid (TA) on Fe bioavailability from Fe supplied as reconstituted ferritin were compared with FeSO4 using an in vitro digestion–Caco-2 cell model. Horse spleen apoferritin was chemically reconstituted into an animal-type ferritin (HSF) and a plant-type ferritin (P-HSF) according to the typical ratios of Fe:P found in these molecules. In the presence of AA (Fe:AA molar ratio of 1:20), significantly more Fe was absorbed from FeSO4 (about 303 %), HSF (about 454 %) and P-HSF (about 371 %) when compared with ferrous sulfate or ferritin without AA. Phytic acid (PA; Fe:PA molar ratio of 1:20) significantly reduced Fe bioavailability from FeSO4 (about 86 %), HSF (about 82 %) and P-HSF (about 93 %) relative to FeSO4 and the ferritin controls. Treatment with TA (Fe:TA molar ratio of 1:1) significantly decreased Fe bioavailability (about 97 %) from both FeSO4 and the ferritin samples. AA was able to partially reverse the negative effect of PA (Fe:PA:AA molar ratio of 1:20:20) on Fe bioavailability but did not reverse the inhibiting effect of TA (Fe:TA:AA molar ratio of 1:1:20) on Fe bioavailability from ferritin and FeSO4. Overall, there were no significant differences in bioavailable Fe between P-HSF, HSF or FeSO4. Furthermore, the addition of AA (a known promoter) or the inhibitors, PA and TA, or both, did not result in significant differences in bioavailable Fe from ferritin relative to FeSO4. The results suggest that Fe in the reconstituted ferritin molecule is easily released during in vitro digestion and interacts with known promoters and inhibitors.

Vegetables may protect against colorectal cancer (CRC) via changes in gene expression involved in anticarcinogenic mechanisms. There is considerable evidence that aberrant DNA methylation plays an important role in carcinogenesis. Furthermore, DNA methylation can be affected by dietary components. Therefore, in the present study, we investigated the DNA methylation status of CpG dinucleotides within the promoter region of the four genes protein kinase C β 1, ornithine decarboxylase 1, fos proto-oncogene and 5,10-methylenetetrahydrofolate reductase in the colon of female sporadic adenoma patients and healthy controls. These genes were chosen because their expression was modulated in response to altered vegetable intake, they are functionally relevant for CRC; they have CpG islands in their promoter region, and a methylation-specific restriction enzyme is available to permit quantitative assay. No significant differences in extent of methylation in colon DNA were detected for any of the four genes in both adenoma polyp patients and healthy controls after altering vegetable intake. Interestingly, before the intervention, ornithine decarboxylase 1 promoter methylation was lower in the colonic mucosa of the adenoma polyp patients when compared with healthy control subjects, which may explain the increased ornithine decarboxylase 1 activity in CRC reported in the literature. In conclusion, we found no evidence that changes in promoter methylation were responsible for differences in expression of four genes in the human colonic mucosa in response to altered vegetable intake. The mechanism(s) responsible for this altered gene expression and, indeed, potential effects on methylation of other genes remain to be determined.

The aim of the present study was to examine the effects of cobalamin (Cbl) on the activity and expression of l-methylmalonyl-CoA mutase (MCM) in rat liver and cultured COS-7 cells. The MCM holoenzyme activity was less than 5 % of the total (holoenzyme+apoenzyme) activity in the liver although rats were fed a diet containing sufficient Cbl. When weanling rats were maintained on a Cbl-deficient diet, the holo-MCM activity became almost undetectable at the age of 10 weeks. In contrast, a marked increase in the total-MCM activity occurred under the Cbl-deficient conditions, and at the age of 20 weeks it was about 3-fold higher in the deficient rats than in the controls (108 (sd 14·5) v. 35 (sd 8·5) nmol/mg protein per min (n 5); P < 0·05). Western blot analysis confirmed that the MCM protein level increased significantly in the Cbl-deficient rats. However, the MCM mRNA level, determined by real-time PCR, was rather decreased. When COS-7 cells were cultured in a medium in which 10 % fetal bovine serum was the sole source of Cbl, holo-MCM activity was barely detected. The supplementation of Cbl resulted in a large increase in the holo-MCM activity in the cells, but the activity did not exceed 30 % of the total-MCM activity even in the presence of Cbl at 10 μmol/l. In contrast, the total-MCM activity was significantly decreased by the Cbl supplementation, indicating that Cbl deficiency results in an increase in the MCM protein level in COS-7 cells as well as in rat liver.

Indices for lean and fat mass adjusted for height derived from bioelectrical impedance for children aged 7 years have been published previously and their usefulness in the clinical assessment of undernutrition has been demonstrated. However, there is a need for norms that cover a wider age range and to explore their functional significance. The aim of the present study is to derive lean and fat indices for children aged 7–11 years and investigate associations with objective measures of cardio-respiratory fitness and grip strength. Subjects were 9574 children participating in the Avon Longitudinal Study of Parents and Children (ALSPAC). Bioelectrical impedance analysis (BIA) data collected longitudinally between ages 7 and 11 were used to derive lean and fat indices using the method of standardised residuals. Cardio-respiratory fitness (CRF) (9 years) and grip strength (11 years) were also measured. Correlation coefficients and 95 % CI were calculated to assess the strength of association between lean index, fat index and CRF and grip strength. Equations for calculating lean and fat indices in children aged 7–11 years relative to the ALSPAC population are presented. Lean index was linearly associated with CRF (rboys 0·20 (95 % CI 0·15, 0·25), rgirls 0·26 (95 % CI 0·22, 0·30)) and grip strength (rboys 0·29 (95 % CI 0·26, 0·32), rgirls 0·26 (95 % CI 0·23, 0·29)). BMI showed slightly weaker associations, while fat index was unrelated to either CRF or grip strength. Lean indices relate to muscle function and fitness while fat index does not.

The aim of the present research was to study if the β-blocker propranolol, which is known to increase bone mass, could reverse the adverse skeletal effects of mild chronic food restriction in weanling rats. Male Wistar rats were divided into four groups: control, control+propranolol (CP), nutritional growth retardation (NGR) and nutritional growth retardation+propranolol (NGRP). Control and CP rats were fed freely with the standard diet. NGR and NGRP rats received, for 4 weeks, 80 % of the amount of food consumed by the control and CP rats, respectively. Results were expressed as mean values and sem. Food restriction induced detrimental effects on body and femur weight and length (P < 0·05) and bone structural and geometrical properties (P < 0·001), confirming results previously shown in our laboratory. However, the β-blocker overcame the deleterious effect of nutritional stress on load-bearing capacity, yielding load, bone stiffness, cross-sectional cortical bone area and second moment of inertia of the cross-section in relation to the horizontal axis without affecting anthropometric, histomorphometric and bone morphometric parameters. The results suggest that propranolol administration to mildly chronically undernourished rats markedly attenuates the impaired bone status in this animal model of growth retardation.

The present results are the last part of a series of experiments conducted in this laboratory on metabolism of major individual rumen ciliate protozoa species and their combinations in sheep. These species were inoculated into the rumen of fauna-free (FF) sheep to study digestibility of dietary fibre and duodenal flow of N. Sixteen rumen- and duodenum-cannulated FF sheep were divided into four equal groups and used in the present experiment. One group remained FF, while the other three were each inoculated intraruminally with the Holotrich protozoa sp. (HT), Entodinium sp. (EN) or total mixed fauna (TF). In comparison with the FF group, the HT did not affect (P>0·05) the duodenal flow of non-NH3-N and bacterial-N, while the flow was decreased (P < 0·05) by both the EN and TF populations. The digestibility of fibre was increased (P < 0·05) by both the EN and TF populations, but the results for the HT group were mostly similar to those for the FF sheep. The fibre digestibility in the stomach for TF was intermediate between that for EN and FF. It was concluded that except when fed high-carbohydrate diets, the presence in the rumen of the HT protozoa are of no significant value or major detriment to ruminant production.

Hydroxytyrosol acetate (HT-AC) is a polyphenol present in virgin olive oil (VOO) at a proportion similar to hydroxytyrosol (HT) (160–479 μmol/kg oil). The present study was designed to measure the in vitro platelet antiaggregating activity of HT-AC in human whole blood, and compare this effect with that of HT and acetylsalicylic acid (ASA). The experiments were designed according to the standard procedure to investigate the activity of ASA. HT-AC and HT inhibited platelet aggregation induced by ADP, collagen or arachidonic acid in both whole blood and platelet-rich plasma (PRP). ASA and HT-AC had a greater effect in whole blood than in PRP when ADP or collagen was used as inducer. ASA and HT-AC had a greater effect in PRP+leucocytes than in PRP alone. All three compounds inhibited platelet thromboxane B2 and leucocyte 6-keto-prostaglandin F1α (6-keto-PF1α) production. The thromboxane/6-keto-PGF1α inhibition ratio (as an indirect index of the prostanoid equilibrium) was 10·8 (se 1) for HT-AC, 1·0 (se 0·1) for HT and 3·3 (se 0·2) for ASA. All three compounds stimulated nitric oxide production, although HT was a weaker effect. In our experiments only concentrations higher than 500 μm (HT) or 1 mm (HT-AC and ASA) inhibited 3-nitrotyrosine production. All three compounds inhibited the production of TNFα by leucocytes, with no significant differences between them. In quantitative terms HT-AC showed a greater antiplatelet aggregating activity than HT and a similar activity to that of ASA. This effect involved a decrease in platelet thromboxane synthesis and an increase in leucocyte nitric oxide production.