An experiment was carried out to evaluate the short-term effect of supplementing a nucleotide-rich yeast extract (NRYE) on growth performance, gut structure, immunity and microflora of piglets raised under sanitary and unsanitary conditions. A total of 84, 21-day old piglets were used in this study; 42 piglets were raised in a room designated as the clean room that was washed once per week, whereas the other 42 piglets were raised in a room designated as the unclean room in which 7 kg of manure from the sow herd was spread on each pen floor on day 1 and 7 and the room was not washed throughout the experiment. The pigs were fed a corn–soybean meal-based diet without or with 0.1% NRYE. Each treatment had 7 replicate pens in each room, and each pen housed 3 pigs. Feed disappearance and BW were recorded on day 1 and 14. On day 14, one pig per pen was euthanized to collect ileum, mesenteric lymph nodes and spleen tissues, and cecum and colon digesta. Overall, NRYE supplementation did not affect growth performance in both clean and unclean conditions, improved kidney weight in both clean (P=0.0002) and unclean room (P<0.0001) and tended to improve the villus height/crypt depth ratio in the clean room (P=0.073). Supplementing NRYE was associated with upregulation of Ileal programmed cell death gene-1 (P=0.0003), interleukin (IL)-1β (P<0.0001), IL-6 (P=0.0003), IL-10 (P<0.0001) and tumor necrosis factor-α (TNF-α) (P<0.0001) in pigs raised in the unclean room. Supplementing the NRYE in pigs raised in the clean room suppressed growth of cecal Enterobacteriacea (P<0.0001) members and colonic Enterococcus spp. (P<0.019), improved proliferation of cecal Lactobacillus spp. (P<0.002) and colonic Clostridium cluster IV (P<0.011) and XVIa members (P<0.0002). Supplementing the NRYE in the unclean room improved proliferation of cecal Clostridium cluster IV (P<0.026) and suppressed proliferation of colonic Enterococcus spp. (P<0.037). In conclusion, supplementing the NRYE to piglets under unsanitary conditions improved ileal immune response by upregulating inflammatory cytokines, and positively modulated proliferation of beneficial gut bacteria and suppression of harmful ones in both clean and unclean rooms.

Trypanosoma evansi is the aetiological agent of trypanosomosis in domestic animals. In this pathology, an inflammatory response can be observed and, as a consequence, the increase of extracellular adenine nucleotides such as ATP. These nucleotide concentrations are regulated by ectoenzymes such as NTPDase (EC 3.6.1.5, CD39), which catalyses the hydrolysis of ATP and ADP into AMP. In this study, the activity of NTPDase in lymphocytes of rats experimentally infected with T. evansi was evaluated. The animals were inoculated with the parasite and monitored by blood smear on a daily basis. The animals were then were divided into 4 groups according to the degree of parasitaemia and period of infection. The blood collections for enzyme analysis and lymphocyte count were performed on the 3rd (beginning of infection), 5th (acute infection) and 15th (chronic infection) days post-infection (p.i.). The control group was composed of non-infected animals. In the infected group a decrease in ATP hydrolysis (36%) was observed on the 3rd day p.i. and a decrease in ADP hydrolysis (62%) was observed on the 5th day p.i. when compared to the control. On the 15th day p.i., an increase in ATP (94%) and ADP (50%) hydrolysis was observed in the infected group. Considering these data it is suggested that NTPDase activity is altered on the surface of lymphocytes of rats infected with T. evansi at different time-points of infection.

The transition from liquid to solid feed during weaning results in morphological, histological and microbial changes in the young animal's intestinal tract and often is associated with diarrhoea. The ban of in-feed antibiotics in pig production in the European Union has led to increasing interest in alternatives to overcome weaning-associated problems. Among others, nucleotides may have the potential to alleviate health impairments due to weaning. Nucleotides are natural components of the non-protein fraction of milk and have important effects on the maintenance of health in young animals. Nucleotides and their related metabolic products play key roles in many biological processes and become essential dietary components when endogenous supply is insufficient for normal function. The present review summarises nucleotide composition of milk from different species, the biology of nucleotides and possible effects of dietary nucleotides on intestinal morphology and function, intestinal microbiota, immune function, nutrient metabolism, hepatic morphology and function as well as growth performance. Special attention is given to data available for pigs, and suggestions are made for inclusion of nucleotides in the diet to benefit piglets' health and reduce the consequences accompanying early weaning.

A combined interdisciplinary research strategy is even more crucial in immunology than in many other biological sciences in order to comprehend the closely linked interactions between cell proliferation, molecular signalling and gene rearrangements. Because of the multi-dimensional nature of the immune system, an abundance of different experimental approaches has developed, with a main focus on cellular and molecular mechanisms. The role of metabolism in immunity has been underexplored so far, and yet researchers have made important contributions in describing associations of immune processes and metabolic pathways, such as the central role of the l-arginine pathway in macrophage activation or the immune regulatory functions of the nucleotides. Furthermore, metabolite supplement studies, including nutritional administration and labelled substrates, have opened up new means of manipulating immune mechanisms. Metabolic profiling has introduced a reproducible platform for systemic assessment of changes at the small-molecule level within a host organism, and specific metabolic fingerprints of several parasitic infections have been characterized by 1H NMR spectroscopy. The application of multivariate statistical methods to spectral data has facilitated recovery of biomarkers, such as increased acute phase protein signals, and enabled direct correlation to the relative cytokine levels, which encourages further application of metabolic profiling to explore immune regulatory systems.

Human milk is recommended as the only alimentary source for the first six months of life. Additionally there is a medical and social need for safe and effective alternative forms of nutrition for infants who cannot be fed with breast milk. Recently the safety and efficacy of some ingredients in infant formulae, such as nucleotides have been discussed. This systematic review analyzed the available evidence to establish the efficacy, safety and dose-response effect of ribonucleotide-supplemented infant formulae (RSIFs). Randomised controlled clinical trials (RCTs) comparing RSIFs to formulae without nucleotides or breast milk were considered in this review. Outcome measures were: antibody titres to common paediatric vaccinations, total lymphocytes, lymphocyte subclasses and NK-cells, episodes of diarrhoea and acute respiratory infection. Publication quality was determined using Jadad and CONSORT guidelines. Results were combined using a random effects model and reported through standardised mean differences (WMD) or risk ratio (RR). Systematic review and meta-analysis showed that RSIFs were associated with a better antibody response to immunisation with Haemopillus influenzae vaccine [SMD 1·74 (99 %CI 1·43–2·05), P = 0·001], diphtheria toxoid [SMD 0·94 (0·75–1·12), P = 0·001], oral polio vaccine [SMD 0·73 (0·51–0·95), P = 0·001], and fewer episodes of diarrhoea [RR 0·67 (0·58–0·76), P = 0·02]. We did not find a major risk of upper respiratory infections [RR 1·11 (0·90–1·36), P = 0·50]. Available evidence suggests a positive benefit of RSFIs on infant health without any risk. These benefits begin with nucleotide addition of 1·9 mg/418·4 kJ and are maintained or increased with 10·78 mg/418·4 kJ.

Recent findings on the presence of water on Mars (Baker, V.R. (2006). Geomorphological evidence for water on Mars. Elements2(3), 139–143; DeJong, E. (2006). Geological evidence of the presence of water on Mars. Abstracts from the 40th Western Regional Meeting of the American Chemical Society, Anaheim, CA, January, 2006, pp. 22–25. American Chemical Society, Washington, DC; McSween, H.Y. Jr. (2006). Water on Mars. Elements2(3), 135–137; Mitrofanov, I.G. (2005). Water explorations on Mars. Priroda9, 34–43) strongly suggest that there existed a period of chemical evolution eventually leading to life processes on primitive Mars (Kanavarioti, A. & Maneinelli, R.L. (1990). Could organic matter have been preserved on Mars for 3.5 billion years. Icarus84, 196–202). Owing to the adverse conditions, it is quite likely that the process of chemical evolution would have been suppressed and any living organisms that formed would have become extinct over time on Mars. The presence of water as a necessity for the survival of living organisms and the presence of grey haematite, originated under aqueous conditions, have led us to investigate the possible role of haematite in the chemical evolution on Mars. Our observations suggest that iron oxide hydroxide (FeOOH), a precursor of haematite, has a much higher binding affinity towards ribose nucleotides (the building blocks of RNA) than the haematite itself. This would mean that during the process of haematite formation, especially through the probable process of Fe3+ hydrolysis by aqueous ammonia, the precursors of haematite might have played a significant role in the processes leading to chemical evolution and the possible origin of life on Mars.

Human milk is a rich source of RNA, free nucleotides (NT) and nucleosides (NS). To determine the uptake of different NS sources by the intestinal epithelium, jejunal explants from suckling piglets were cultured in a medium supplemented with a mixture of NS (adenosine, cytidine, guanosine, inosine, uridine; 10 mg/l each), a mixture of five NT (AMP, CMP, GMP, IMP, UMP; 7 mg/l each) or RNA (60 mg/l), respectively. Aliquots from the media were taken at different times (0·5, 2, 5, 15, 30, 60, 180 min). NS and NT concentrations were analysed in the different supernatants at those periods using solid-phase extraction followed by HPLC. When explants were cultured in the presence of NS the concentration of these compounds, excepting cytidine, rapidly decreased, suggesting that they are efficiently taken up. When explants were incubated in the presence of NT, the total concentration of these compounds decreased while the total concentration of NS increased, suggesting that enterocytes efficiently hydrolyse NT into NS. Likewise, when explants were incubated in the presence of RNA, the total concentration of both NT and NS increased, indicating that intestinal explants are able to hydrolyse RNA to NT and then to NS in the absence of luminal enzymes. In conclusion, the jejunum of piglets at weaning is able to hydrolyse RNA and free NT to NS, and NS, excepting cytidine, are efficiently taken up by the small intestine. These results suggest that the current concentration of NT used to supplement infant formulas should be reconsidered.

The objective of the present study is to demonstrate the effect of nucleotide intake and intensive nutritional support on the concentration of insulin-like growth factor I (IGF-I) and other hormonal biomarkers in severely malnourished children. Twenty-six severely malnourished children <48 months of age received formula without lactose via enteral feeding for 2 weeks and ad libitum for an additional 2 weeks. Anthropometrical measurements were performed and serum concentrations of IGF-I, insulin-like growth factor binding protein-3 (IGFBP-3), leptin, soluble leptin receptor (sOB-R), as well as the estimated molar excess of sOB-R over leptin were obtained. Two groups were formed. One group received formula with nucleotides (NT+; n 13) and the other without nucleotides (NT−; n 13). A control group was included (n 13). Parametric and non-parametric tests as well as ANOVA models were used. Nutritional recovery, nucleotides intake, type of malnutrition, age and the interaction between gender and malnutrition influenced the concentration of IGF-I (P<0·001). Nutritional recovery, nucleotides intake, gender and type of malnutrition had an effect on IGFBP-3 (P<0·001). Nutritional recovery had a significant effect on serum leptin (P=0·001). Age and nutritional recovery had an effect on sOB-R (P<0·001); all variables included affected the molar excess of sOB-R over leptin (P<0·001). In conclusion, nucleotide intake and nutritional recovery had a notable effect on IGF-I, IGFBP-3 and other hormonal biomarkers. This outcome could stimulate the catch-up growth of severely malnourished infants and toddlers during the nutritional recovery period.

The aim of the present study was to evaluate the effect of nucleotide supplementation on the oxidative stress induced by a high proportion of dietary polyunsaturated fatty acids ( PUFAs) in pigs. Twenty-four male growing pigs were penned individually and after an adaptation period divided into three groups. All groups received isocaloric daily rations composed of a basal diet supplemented with either: starch (CONT), linseed oil (LIN) and LIN and nucleotides (LIN + NUC). The experimental period lasted 21 days. Oxidative stress was evaluated by measuring the degree of lymphocyte nuclear DNA damage, the urine malondialdehyde ( MDA) excretion rate, erythrocyte glutathione peroxidase concentration and the total anti-oxidant status of plasma. Malondialdehyde concentrations in the blood and MDA urinary excretion rates were higher (P < 0·01) in animals supplemented with LIN and LIN + NUC compared with CONT animals. The degree of DNA damage in the LIN-supplemented animals was also higher (P < 0·01). Compared with the LIN-supplemented animals, nucleotide supplementation reduced (P < 0·01) the degree of DNA damage in lymphocytes to the level of the CONT group. Erythrocyte glutathione peroxidase concentration and plasma total anti-oxidant status were similar across treatments. The results of this experiment indicate that nucleotide supplementation effectively eliminates the genotoxic effects of high PUFA intakes on blood lymphocytes and demonstrates new evidence for the immunonutritive effect of nucleotides.

The composition of fourteen infant formulae and six follow-up milks with regard to their free amino acids (including taurine), free nucleotides, orotic acid, and free and total L-carnitine content was studied. The levels found were compared with the limits established in European legislation and with the composition of human and cows' milk samples. HPLC methodologies, optimized and validated for the matrices under study, were used, except for free and total L-carnitine contents that were quantified using a flow-injection manifold, also optimized and validated for the matrices under study. Global statistical treatment of the results by cluster analysis indicated similarities between the contents of the N compounds under study of infant formulae, follow-up milks and cows' milk and differences with regard to human milk composition. The principal component analysis showed that 60·2% of the variation in data was due to the first principal component, and the second component represented 23·8% of the total information. Nucleotide profiles, orotic acid, and free and total L-carnitine contents explain the main differences observed between human milk and the other milks studied (cows' milk, infant formulae and follow-up milks). Cows' milk is distinguished from infant formulae and follow-up milks mainly owing to the different uric acid contents and free amino acids profiles.

This study examined the effects of glutamine and nucleotides on growth, intestinal villous height and immune responses of weaned pigs. Eighty weaned pigs (Landrace ✕ Yorkshire ✕ Duroc) were randomly assigned to five treatments, including a control diet and a 2 (glutamine 1·0 and 1·5%) by 2 (nucleotides 500 and 1000 p.p.m.) factorial design. There were no differences among the five treatments in weight gain from weeks 0 to 4 and 0 to 8 of the experiment. However, the food intake of treatment 3 (glutamine 1·0% and nucleotides 1000 p. p. m. ) for weeks 4 to 8 was higher (P < 0·05) than that of the control group. Intestinal villous height, and serum immunoglobulin-G concentration 2 h post lipopolysaccharide injection, were higher (P < 0·05) for pigs administered treatment 3 than for controls. Furthermore, foot and mouth disease (FMD) neutralizing antibody titres were also higher (P < 0·05) than the control group for pigs given treatment 3 when measured 3 weeks following vaccination with FMD antigen. These observations suggest that a combination of 10% of glutamine and 1000 p. p. m. of nucleotides in the diet could improve food intake and intestinal villous height and promote the immune responses and FMD antibody titres of weaned pigs.

The nucleotides and nucleoside diphosphate monosaccharides (NDP-sugars), UDP-galactose, UDP-glucose, UDP, ATP, ADP, ADP-glucose, AMP and NAD, some of which are involved in carrageenan biosynthesis, were identified in the two carrageenophytes Calliblepharis jubata and Solieria chordalis and quantified by high-performance liquid chromatography (HPLC). Of several extraction procedures evaluated, a freeze-thaw step followed by formic acid (1 M) extraction was the preferred method. The pool size of nucleotides and NDP-sugars differed between the two algal species, which may be related to their growth state. Calliblepharis, which was not growing actively, had larger pools than Solieria, which was in an active growth phase. Using an HPLC method UDP-D-glucose-4-epimerase activity was measured in cell-free extracts of both algae, for the first time in a carrageenophyte.

Adenylate Energy Charge (AEC) values of Crassostrea gigas (9 and 18 months old) cultured in ponds in Marennes-Oléron Bay (Atlantic coast, France) are reported. Two ponds were loaded with two different initial oyster biomasses in order to obtain different food conditions. The growth of oysters was different in the two ponds which corroborated the effectiveness of attaining different food conditions. AEC level was different depending on the pond and was therefore related to food availability per unit oyster biomass. Well-fed oysters exhibited greater AEC values than poorly-fed oysters, throughout the study. Information on nutritional situations for wild or cultured populations can therefore be obtained by AEC. In each pond, the seasonal AEC variations were similar for both age-groups and were characterized by a minimum value in summer and a maximum value in winter. The influence of reproduction on these variations cannot be ruled out as for all the batches maturation processes were evidenced. Research will be done on a temperature effect to try to distinguish these seasonal variations. A strong correlation between adenylate energy charge (AEC) and guanylate energy charge (GEC) was demonstrated; it was similar for both ponds. The levels of nucleotide concentration were relatively stable during the survey except in September and December for oysters in the low density pond. Among nucleotides, adenylate pool represented the major part (80%). A decrease in the total concentration of adenylates was observed in August. The best growth correlated with the lowest adenylate percentage and a better GTP/ATP ratio, suggesting that this ratio could be a useful index for growth.