β-Cryptoxanthin (β-CX), a provitaminic carotenoid of potential interest for health, is found principally in Citrus fruit in both free and esterified forms. Little is known about the intestinal absorption of β-CX especially with regard to the esterified forms. The aim of this study was to evaluate the absorption of free and esterified β-CX using simulated digestion coupled with the Caco-2 model. Bioaccessibility was investigated by measuring the transfer of carotenoids from different citrus juices into micelles using an in vitro digestion system. Then, carotenoid uptake was evaluated by adding carotenoid-rich micelles (from the in vitro digestion) or synthetic micelles (made from synthetic lipids and carotenoids purified from citrus juice) to human intestinal cells (Caco-2 TC7 clone). Our results showed that β-cryptoxanthin esters (β-CXE) were partially hydrolysed during the in vitro digestion. The bioaccessibility of free β-CX measured was significantly higher (40 (sd 1·05) %) than that of β-carotene (30 (sd 1·9) %) and β-CXE (16 (sd 1·5) %). In the same way, the incorporation of free β-CX (27 (sd 1·01) %) into synthetic micelles exceeded (P < 0·05) that of β-carotene (10 (sd 0·7) %) and β-CXE (8·8 (sd 0·4) %). In the case of micelles from in vitro digestion, the uptake of β-carotene, free β-CX and β-CXE forms by Caco-2 cells was 14·3 (sd 1·8), 3·9 (sd 1·3), and 0·7 (sd 0·08) % respectively. These results showed a preferential uptake by Caco-2 cells of β-carotene and free β-CX compared with the two esters of β-CX.

Dietary soya protein substitution for casein initiated at weaning slows disease progression in animal models of chronic renal disease. As there is increasing evidence that fetal programming can have a significant impact on kidney physiology and function in offspring, the objective of the current study was to determine whether exposure to soya protein in the diet earlier than weaning would have further benefits. Han:SPRD-cy (cy/+) breeder rats were fed a casein-based or soya protein-based diet 2 weeks prior to mating, throughout pregnancy and during lactation. Following this maternal period, 3-week-old pups were given either the same or the alternate diet for a 7-week weaning period. Dietary soya protein compared with casein in the maternal or weaning period both independently resulted in less renal inflammation (macrophage infiltration lower by 24 % (P = 0·0003) and 32 % (P < 0·0001), respectively). When soya protein was given in both feeding periods, the effect was additive. Soya protein substitution for casein resulted in less oxidative damage as indicated by 28 % lower oxidized-LDL staining (P = 0·013) when present in the maternal period, or in the weaning period (by 56 %, P < 0·0001). Renal cell proliferation was reduced by 29–33 % (P < 0·05) in rats given soya protein whether the exposure was during the maternal or weaning period. Soya protein compared with casein in the maternal period also resulted in 33 % (P = 0·0013) less proteinuria, indicating superior renal function. Dietary soya protein during pregnancy and lactation represents a potential preventative approach in treating for those with congenital kidney diseases.

The methionine cycle and methyl group metabolism are implicated in the long-term programming of metabolism. Diets deficient in folic acid, methionine and choline have been fed to pregnant rats to examine the effects on amino acid metabolism, choline reserves and DNA methylation in dam and fetuses. Animals were fed folate-deficient, folate-deficient with low methionine, folate-deficient with low choline and folate-deficient, low-methionine, low-choline diets starting 2 weeks before mating. The dams and their fetuses were subsequently killed on day 21 of gestation for analysis. Diets low in methionine reduced fetal and maternal weight. Folate deficiency increased the concentrations of homocysteine, glycine, serine and threonine in the maternal plasma, and this was exacerbated by the low-methionine diets. The changes in the amino acid profile in the fetal serum were similar but less pronounced. This result suggests that fetal metabolism was less perturbed. Folate deficiency increased free choline in the maternal liver at the expense of phosphocholine stores. It has been suggested that a deficiency in methyl donors in the diet during pregnancy may impact on key methylation reactions, including the methylation of DNA. Despite widespread changes in the metabolism of choline and amino acids, there was no change in the global methylation of cytosine in DNA from either maternal or fetal livers. This suggests a more indirect mechanism in which gene–nutrient interactions modify the process of differential methylation during development

Forty-eight, 21-d-old pigs were used to examine the effects of different types of cooked white rice on starch digestion, digesta and fermentation characteristics, shedding of β-haemolytic Escherichia coli and performance after weaning. Pigs received one of three rice-based diets: (i) medium-grain Amaroo (AM), (ii) long-grain Doongara (DOON), and (iii) waxy (WAXY). The remainder of the diet consisted predominantly of animal proteins. A fourth diet contained mainly wheat, barley and lupins (WBL). On days 1, 3, 7 and 9 after weaning, a faecal swab was taken for assessment of β-haemolytic E. coli and faecal consistency. Apparent digestibility of starch measured in the ileum 14 d after weaning was highest (P = 0·004) in AM and WAXY and lowest, but the same (P>0·05), in DOON and WBL. Starch digestibility in the rectum was highest in all rice diets (P < 0·001). Digesta viscosity was highest in pigs fed WBL in both the ileum (P < 0·001) and caecum (P = 0·027). Pigs fed rice generally had lighter (P < 0·05) gastrointestinal organs than pigs fed WBL. Performance of pigs was similar for all treatments; however, pigs fed rice-based diets had a higher (P < 0·001) carcass percentage than pigs fed WBL. Pigs fed WBL produced more acid (P < 0·05) but had lower molar proportions of acetate (P < 0·05), isobutyrate (P < 0·01) and isovalerate (P < 0·001) and a higher molar proportion of butyrate (P < 0·01) in the large intestine than pigs fed rice. Shedding of E. coli was low; however, pigs fed AM and WBL shed less E. coli than pigs fed other diets.

Recent cross-sectional data suggest that better vitamin K status in young girls (aged 3–16 years) is associated with decreased bone turnover, even though it is not associated with bone mineral content (BMC). The objective of the present study was to investigate the relationship between serum percentage of undercarboxylated osteocalcin (%ucOC), as an index of vitamin K status, and BMC and biochemical indices of bone turnover in peri-pubertal Danish girls. This peri-pubertal stage is a dynamic period of bone development, and as such, may represent an important window of opportunity for vitamin K status to modulate childhood bone health. Serum %ucOC and serum 25-hydroxyvitamin D (25 (OH) D) were measured at baseline in a study of 223 healthy girls aged 11–12 years. Urinary pyridinium crosslinks of collagen and serum total osteocalcin as markers of bone resorption and formation, respectively, as well as BMC (total body and lumbar spine) were also measured. Serum %ucOC (median 21·9 %) was not associated with markers of bone resorption or with total osteocalcin. Serum %ucOC was inversely correlated with serum 25 (OH) D (r − 0·143; P < 0·05). Serum %ucOC was negatively associated with BMC of the total body (β − 0·045; P < 0·001) and lumbar spine (β − 0·055; P < 0·05), after adjustment for potential confounders including vitamin D status. Better vitamin K status was associated with increased BMC, but not bone turnover, in healthy peri-pubertal Danish girls. There is a need for well-designed, randomized phylloquinone supplementation trials in children and adolescents to confirm epidemiological findings of an association between vitamin K status and bone health.

The metabolism of flavone C-glucosides and p-coumaric acid from antioxidant of bamboo leaves (AOB) in rats is discussed systematically in the present study. Following single oral administration of AOB, p-coumaric acid was detected in plasma but not in gastrointestinal tract extracts and faeces, and the corresponding absorption pharmacokinetic curve at different time points showed a prolonged elimination phase with p-coumaric acid being detected in the kidneys and excreted as its original form (1·80 (sd 0·24) % and 1·90 (sd 0·26) % at 12 and 24 h, respectively). However, the four flavone C-glucosides orientin, homoorientin, vitexin and isovitexin were poorly absorbed in the gastrointestinal tract. More than 50 % recovery of flavone C-glucosides was determined at 12 h and faeces containing these four analytes (21·23 (sd 1·92) %) were excreted at 24 h. These data suggested that the effective time these compounds were in the colon was long enough so that they could exert their antioxidant activity and scavenge free radicals. Besides the excretion of the original forms, moieties of the flavone C-glucosides were hydrolysed by deglycosylation and the opening of the heterocyclic C ring. Some small molecules such as phloroglucinol (PG), hydrocaffeic acid (HCA) and phloretic acid (PA) were detected and identified as metabolites of the flavone C-glucosides. In the present work, we compared the metabolic fate of flavone C-glucosides to that of flavone O-glucosides in rats, and evaluated the absorption, tissue distribution and excretion of flavone C-glucosides in AOB on their metabolism for the first time.

The effects of asynchronous availability of amino acids and glucose on muscle composition and enzyme activities in skeletal muscle were studied in preruminant calves. It was hypothesized that decreased oxidative enzyme activities in muscle would explain a decreased whole body heat production with decreasing nutrient synchrony. Preruminant calves were assigned to one of six degrees of nutrient synchrony, step-wise separating the intake of protein and lactose over the two daily meals. Calves at the most synchronous treatment received two identical meals daily. At the most asynchronous treatment, 85 % of the daily protein and 20 % of the daily lactose supply were fed in one meal and the remainder in the other meal. Daily intakes of all dietary ingredients were identical for all treatments. Oxidative enzyme activities and fat content increased with decreasing nutrient synchrony in M. Rectus Abdominis (RA), but not in M. Semitendinosus. Cytochrome-c-oxidase activity was positively correlated with fat content in RA (r 0·49; P < 0·01). Oxidative enzyme activities in both muscles were not correlated with average daily heat production, but citrate synthase activity in RA was positively correlated (P < 0·01) with the circadian amplitude (r 0·53) and maximum (r 0·61) of heat production associated with physical activity. In conclusion, this study indicates that muscle energy stores are regulated by nutrient synchrony. The lack of correlation between muscle oxidative enzyme activities and average daily heat production was in contrast with findings in human subjects. Therefore, oxidative enzyme activity in muscle should not be used as an indicator for whole body heat production in growing calves.

Effects of sesamin and sesamolin (sesame lignans) on hepatic fatty acid metabolism were compared in rats. Rats were fed either a lignan-free diet, a diet containing 0·6 or 2 g/kg lignan (sesamin or sesamolin), or a diet containing both sesamin (1·4 g/kg) and sesamolin (0·6 g/kg), for 10 d. Sesamin and sesamolin dose-dependently increased the activity and mRNA abundance of various enzymes involved in hepatic fatty acid oxidation. The increase was much greater with sesamolin than with sesamin. These lignans increased parameters of hepatic fatty acid oxidation in an additive manner when added simultaneously to an experimental diet. In contrast, they decreased the activity and mRNA abundance of hepatic lipogenic enzymes despite dose-dependent effects not being necessarily obvious. Sesamin and sesamolin were equally effective in lowering parameters of lipogenesis. Sesamolin accumulated in serum at 33- and 46-fold the level of sesamin at dietary concentrations of 0·6 and 2 g/kg, respectively. The amount of sesamolin accumulated in liver was 10- and 7-fold that of sesamin at the respective dietary levels. Sesamolin rather than sesamin can account for the potent physiological effect of sesame seeds in increasing hepatic fatty acid oxidation observed previously. Differences in bio-availability may contribute to the divergent effects of sesamin and sesamolin on hepatic fatty acid oxidation. Sesamin compared to sesamolin was more effective in reducing serum and liver lipid levels despite sesamolin more strongly increasing hepatic fatty acid oxidation.

The present study was conducted to determine the effects of α-lipoic acid (ALA) on fatty acid mobilisation and ketone body production in female broiler chickens. In the first experiment, chickens received an intravenous infusion of ALA (0, 25 or 50 mg/kg per h) for 90 min. Blood was drawn at 30 min intervals, and plasma glucose, NEFA, free glycerol and hydroxybutyrate were analysed. In the second experiment, ALA (100 mg/kg per h) was continuously infused to chickens fed or fasted for at least 24 h. Changes in plasma metabolites for 90 min were determined. In the first experiment, as shown by the response area, ALA infusion did not affect plasma glucose but increased (P < 0·05) plasma NEFA, regardless of the dose level. In contrast, plasma hydroxybutyrate was reduced at the lower infusion rate (P < 0·05). No significant changes in plasma free glycerol were observed. In experiment 2, ALA stimulated both plasma glucose (P < 0·01) and NEFA (P < 0·001) and the responses were greater than those of controls, regardless of the feeding state. The interaction between ALA and the feeding condition had a significant effect on plasma hydroxybutyrate (P < 0·01). ALA reduced plasma hydroxybutyrate, and this response was greater in the fasted birds than in those fed. Therefore, the present study found a lowering effect of ALA on plasma hydroxybutyrate level and suggests that the ALA-induced plasma NEFA increment was attributable to decreased hepatic fatty acid oxidation in chickens.

One approach to achieve weight loss and decrease both obesity and associated morbidities involves high-protein, low-carbohydrate (HPLC) diets. This study compares the impact on metabolic health of HPLC and high-protein, medium-carbohydrate (HPMC) diets offered to diet-induced obese (DIO) rats. Weanling male rats were fed either a 37 % fat diet (n 48) or stock pellets (n 12) for 22 weeks. Rats fed the 37 % fat diet accumulated more body fat (26·6 versus 14·8 % body weight, P < 0·001) compared with those on stock diet. The DIO rats had higher systolic blood pressure (+6·6 mmHg, P = 0·002), fasting insulin (+63 % P = 0·006) and areas under the glucose (+21 %, P < 0·001) and insulin (+81 %, P < 0·001) curves following an oral glucose tolerance test. DIO rats were then separated into four groups and offered for 8 weeks either: (1) the 37 % fat diet; (2) an HPLC or (3) HPMC diet; or (4) fed the 37 % fat diet to the intake of the HPMC group. Rats offered the 37 % fat or HPLC diets gained while those on HPMC lost body fat. Blood pressure was not altered by the dietary switch. Both HPLC and HPMC rats had lowered fasting insulin (P = 0·027) and improved homeostatic assessment (HOMA; P = 0·011) that was not different from those of stock animals. These improvements occurred despite differences in fat gain, and indicate that both weight loss and macronutrient intake can impact favourably on obesity-associated morbidities.

Protein–energy malnutrition is a syndrome in which anaemia together with multivitamin and mineral deficiency may be present. The pathophysiological mechanisms involved have not, however, yet been completely elucidated. The aim of the present study was to evaluate the pathophysiological processes that occur in this anaemia in animals that were submitted to protein–energy malnutrition, in particular with respect to Fe concentration and the proliferative activity of haemopoietic cells. For this, histological, histochemical, cell culture and immunophenotyping techniques were used. Two-month-old male Swiss mice were submitted to protein–energy malnutrition with a low-protein diet (20 g/kg) compared with control diet (400 g/kg). When the experimental group had attained a 20 % loss of their original body weight, the animals from both groups received, intravenously, 20 IU erythropoietin every other day for 14 d. Malnourished animals showed a decrease in red blood cells, Hb concentration and reticulocytopenia, as well as severe bone marrow and splenic atrophy. The results for serum Fe, total Fe-binding capacity, transferrin and erythropoietin in malnourished animals were no different from those of the control animals. Fe reserves in the spleen, liver and bone marrow were found to be greater in the malnourished animals. The mixed colony-forming unit assays revealed a smaller production of granulocyte–macrophage colony-forming units, erythroid burst-forming units, erythroid colony-forming units and CD45, CD117, CD119 and CD71 expression in the bone marrow and spleen cells of malnourished animals. These findings suggest that, in this protein–energy malnutrition model, anaemia is not caused by Fe deficiency or erythropoietin deficiency, but is a result of ineffective erythropoiesis.

A [13CO2] breath test was applied as a non-invasive method to study the catabolism of ingested amino acids shortly after a meal. This test requires the ingestion of a [1-13C]-labelled amino acid and the analysis of expired air for [13C] enrichment and CO2. The recovery of label as [13CO2] reflects the catabolism of the [1-13C]-labelled substrate. Such a non-steady state approach provides information that is complementary to the information obtained by steady-state methods using a primed continuous infusion of tracer amino acids during the fed state. In a model study with twenty adult male rats, two groups of animals were fed twice a day with one of two semi-synthetic iso-energetic diets. One diet contained egg white protein (EW) as the sole amino acid source. The second diet contained a mixture of free amino acids with a pattern similar to that of the EW diet. On day 5 of the dietary treatment, l-[1-13C]leucine, either bound in EW protein or in free form, was ingested as part of the morning meal. The expired air was sampled at 30 min intervals for 5 h. The rate of recovery ranged from 0 % to 6 % of the dose/h. Up to 120 min after the onset of the meal, the recovery values for the free amino acid diet were higher than those for the EW diet. Differences in recovery reflect differences in postprandial utilisation. The differences in label recovery were mainly determined by the [13C] enrichment of the expired air. As a consequence, CO2 measurements are not mandatory when CO2 production is comparable.

The effects of the type of grass silage and dietary inclusion of fish oil (FO) on the fatty acid profile of bovine intramuscular and subcutaneous adipose tissue were investigated. Eighty Friesian steers were assigned (n 10) to unwilted or wilted silage, and to one of four rations which contained, per kg, 80 g of sunflower oil and either 0, 10, 20 or 40 g of FO replacing lard. Animals were slaughtered after 108 d and the fatty acid profile of the neutral, polar and total lipid fractions of the M. longissimus dorsi, and the total lipid fraction of the subcutaneous adipose tissue were determined. Wilting of grass prior to ensiling increased the concentration of conjugated linoleic acid (CLA) in intramuscular total lipid (P < 0·01), but did not affect the n-6:n-3 PUFA ratio. Increasing FO supply linearly increased (P < 0·05) the concentration of the cis-9, trans-11 and trans-10, cis-12 isomers of CLA and trans-11 18 : 1 predominantly in the neutral lipid fraction of intramuscular total lipid, and linearly decreased the n-6:n-3 PUFA ratio. Wilting of grass prior to ensiling increased the concentration of CLA in subcutaneous adipose tissue (P < 0·001), while increasing FO supply linearly increased the concentration of cis-9, trans-11 CLA. From a human nutrition perspective, increasing the level of FO in the ration or wilting of grass prior to ensiling appear to modify the fatty acid composition of beef muscle favourably. However, the health implications of associated increases in trans fatty acids remain to be elucidated.

Excessive I in the diet of pregnant sheep can reduce the concentration of antibodies in the blood plasma of the lambs after they have consumed colostrum. Our aim was to determine the dose of dietary I that would avoid this effect, and to relate this to changes in the concentrations of hormones and metabolites in the lambs. Four groups of pregnant ewes received concentrate containing 5·5, 9·9, 14·8, and 21·0 mg I/kg DM, respectively. Hay and molasses (containing 0·16 and 0·29 mg I/kg DM, respectively) were available ad libitum. The lambs were prevented from suckling for the first 24 h of life and were fed a fixed quantity of artificial colostrum in four feeds. At 24 h, the average plasma concentrations of immunoglobulin G in the lambs were 6·08, 5·06, 3·18 and 3·10 g/l for the 5·5, 9·9, 14·8 and 21·0 mg/kg groups, respectively. Supplementation with I was associated with higher levels of tri-iodothyronine and thyroxine in the lambs at birth. There was no effect of treatment on the plasma concentrations of insulin, cortisol, glucose or NEFA in the lambs. The concentration of dietary I that had an effect on the immunoglobulin concentration in the lambs is marginally above the levels added to commercial concentrate feeds; we tentatively identify 9·9 mg I/kg DM (approximately 9 mg I/ewe per day) as the upper safe limit of I supplementation according to the criterion of the concentration of immunoglobulin G in the plasma of lambs at 24 h after birth.

The intestinal anti-inflammatory effects of two probiotics isolated from breast milk, Lactobacillus reuteri and L. fermentum, were evaluated and compared in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis. Colitis was induced in rats by intracolonic administration of 10 mg TNBS dissolved in 50 % ethanol (0·25 ml). Either L. reuteri or L. fermentum was daily administered orally (5 × 108 colony-forming units suspended in 0·5 ml skimmed milk) to each group of rats (n 10) for 3 weeks, starting 2 weeks before colitis induction. Colonic damage was evaluated histologically and biochemically, and the colonic luminal contents were used for bacterial studies and for SCFA production. Both probiotics showed intestinal anti-inflammatory effects in this model of experimental colitis, as evidenced histologically and by a significant reduction of colonic myeloperoxidase activity (P < 0·05). L. fermentum significantly counteracted the colonic glutathione depletion induced by the inflammatory process. In addition, both probiotics lowered colonic TNFα levels (P < 0·01) and inducible NO synthase expression when compared with non-treated rats; however, the decrease in colonic cyclo-oxygenase-2 expression was only achieved with L. fermentum administration. Finally, the two probiotics induced the growth of Lactobacilli species in comparison with control colitic rats, but the production of SCFA in colonic contents was only increased when L. fermentum was given. In conclusion, L. fermentum can exert beneficial immunomodulatory properties in inflammatory bowel disease, being more effective than L. reuteri, a probiotic with reputed efficacy in promoting beneficial effects on human health.

Probiotics (PRO) modulate immunity in humans, while the effect of prebiotics (PRE) and synbiotics (SYN) on the human immune system are not well studied yet. The objective of this study was to investigate whether daily intake of a SYN modulates immune functions. In a randomised double-blind, placebo-controlled trial, thirty-four colon cancer patients who had undergone ‘curative resection’ and forty polypectomised patients participated. Subjects of the SYN group daily received encapsulated bacteria (1 × 1010 colony-forming units of Lactobacillus rhamnosus GG (LGG) and 1 × 1010 colony-forming units of Bifidobacterium lactis Bb12 (Bb12)) and 10 g of inulin enriched with oligofructose. Controls received encapsulated maltodextrin and 10 g of maltodextrin. Prior to intervention (T1), and 6 (T2) and 12 weeks after the start of the intervention (T3), phagocytic and respiratory burst activity of neutrophils and monocytes, lytic activity of natural killer cells and production of interleukin (IL)-2, IL-10 and IL-12, as well as tumour necrosis factor-α and interferon-γ (IFN-γ) by activated peripheral blood mononuclear cells (PBMC) were measured. In faeces, the concentrations of transforming growth factor-β1 and prostaglandin E2 were measured. IL-2 secretion by activated PBMC from the polyp group increased significantly between T1 or T2 and T3 (P < 0·05). In the cancer group, SYN treatment resulted in an increased capacity of PBMC to produce IFN-γ at T3 (P < 0·05). Other immunity-related parameters were not affected by SYN treatment, neither in the cancer nor in the polyp group. In conclusion, supplementation with this SYN has minor stimulatory effects on the systemic immune system of the two study groups. Further studies in humans should aim to focus on the gut-associated immune system.

We examined the physiological importance of bovine dietary proteins in rats fed diets prepared from bovine Achilles' tendons and arteries. Rats were fed for 4 weeks, with a 20 % casein diet (CON), in comparison with two diets containing 15 % casein and 5 % of either bovine Achilles' tendon (AC) or artery (AR) protein preparations. The serum total cholesterol concentration and non-HDL-cholesterol level in the AR-fed group were significantly lower (P < 0·05) than those in the CON-fed group at the end of the 4-week feeding period. The hepatic mRNA were measured, and the hydroxyl methyl glutaryl-CoA reductase mRNA level was significantly lower (P < 0·05) in the AR-fed group compared with the CON-fed group. Total hepatic cholesterol concentration in AC-fed rats was significantly (P < 0·05) higher than in the CON-fed group. The serum TAG concentration and fatty acid synthase mRNA level in AC- and AR-fed groups were significantly lower (P < 0·05) compared with the CON-fed group throughout the feeding period. Faecal neutral sterol excretion was significantly (P < 0·05) higher in the AC- and AR-fed groups compared with the CON-fed group. The results of the present study demonstrate that some bovine dietary proteins have similar functions as dietary fibres, lowering serum lipid concentration by enhancing faecal neutral sterol excretion or suppressing lipid synthesis in the liver. Moreover, favourable amino acid compositions in the AR and AC preparations may also have a lowering effect on plasma lipid concentration in bovine protein diet-fed groups.

The aim was to investigate the effects of a 48 h period of fluid, energy or combined fluid and energy restriction on salivary IgA (s-IgA) responses at rest and after exercise. Thirteen healthy males (age 21 (sem 1) years) participated in four randomised 48 h trials. In the control trial participants received their estimated energy (12 154 (sem 230) kJ/d) and water (3912 (sem 140) ml/d) requirements. On fluid restriction (FR) participants received their energy requirements and 193 (sem 19) ml water/d to drink and on energy restriction (ER) participants received their water requirements and 1214 (sem 25) kJ/d. Fluid and energy restriction (F+ER) was a combination of FR and ER. After 48 h, participants performed a 30 min treadmill time trial (TT) followed by rehydration (0–2 h) and refeeding (2–6 h). Unstimulated saliva was collected at 0, 24 and 48 h, post-TT, and 2 and 6 h post-TT. Saliva flow rate (sflw) and s-IgA (ELISA) remained unchanged in control conditions and on ER. However, 48 h on FR decreased sflw (64 %) which most probably accounted for the increase in s-IgA concentration (P < 0·01). Despite a decrease in sflw (54 %), s-IgA concentration did not increase on F+ER, resulting in a decreased s-IgA secretion rate by 24 h (0 h: 20 (sem 2); 24 h: 12 (sem 2) μg/min; P < 0·01). Post-TT s-IgA secretion rate was not lower compared with 48 h on any trial. s-IgA secretion rate returned to within 0 h values by 6 h post-TT on F+ER. In conclusion, a 24–48 h period of combined F+ER decreased s-IgA secretion rate but normalisation occurred upon refeeding.

Several studies have described reduced plasma concentrations of arginine, the substrate for nitric oxide synthase (NOS) in infants with necrotizing enterocolitis (NEC). No information on the plasma concentrations of the endogenous NOS inhibitor asymmetric dimethylarginine (ADMA) in patients with NEC is currently available. We investigated whether plasma concentrations of arginine, ADMA, and their ratio differ between premature infants with and without NEC, and between survivors and non-survivors within the NEC group. In a prospective case–control study, arginine and ADMA concentrations were measured in ten premature infants with NEC (median gestational age 193 d, birth weight 968 g), and ten matched control infants (median gestational age 201 d, birth weight 1102 g), who were admitted to the Neonatal Intensive Care Unit. In the premature infants with NEC, median arginine and ADMA concentrations (μmol/l), and the arginine:ADMA ratio were lower compared to the infants without NEC: 21·4 v. 55·9, P = 0·001; 0·59 v. 0·85, P = 0·009 and 36·6 v. 72·3, P = 0·023 respectively. In the NEC group, median arginine (μmol/l) and the arginine:ADMA ratio were lower in non-surviving infants than in surviving infants: 14·7 v. 33·8, P = 0·01 and 32·0 v. 47·5, P = 0·038 respectively. In premature infants with NEC not only the NOS substrate arginine, but also the endogenous NOS inhibitor ADMA and the arginine:ADMA ratio were lower than in infants without NEC. In addition, low arginine and arginine:ADMA were associated with mortality in infants with NEC. Overall, these data suggest that a diminished nitric oxide production may be involved in the pathophysiology of NEC, but this needs further investigation.

Experimental evidence suggests that most herbs and spices possess a wide range of biological and pharmacological activities that may protect tissues against O2-induced damage. The objectives of the present study were: first, to determine the effects of plant extracts on the viability, membrane integrity, antioxidant status and DNA integrity of Caco-2 cells and second, to investigate the cytoprotective and genoprotective effects of these plant extracts against oxidative stress in Caco-2 cells. The plant extracts examined were rosemary (Rosmarinus officinalis L.), oregano (Origanum vulgare L.), sage (Salvia officinalis L.) and echinacea (Echinacea purpurea L.). Cell membrane integrity was assessed by the lactate dehydrogenase release assay. Viability was determined by the neutral red uptake assay (NRUA) and the concentration of compound that resulted in 50 % cell death (IC50) was calculated. Antioxidant status of the cells was assessed by measuring GSH content, catalase activity and superoxide dismutase activity. To examine their cytoprotective and genoprotective effects, Caco-2 cells were pre-treated with each plant extract for 24 h followed by exposure to H2O2. DNA damage was assessed by the comet assay and cell injury was determined by the NRUA. Rosemary was the most toxic (IC50 123 μg/ml) and echinacea the least toxic (IC50 1421 μg/ml). Sage was the only plant extract to affect the antioxidant status of the cells by increasing GSH content. Sage, oregano and rosemary protected against H2O2-induced DNA damage (olive tail moment and percentage tail DNA), whereas protection against H2O2-induced cytotoxicity was afforded by sage only.