Effect of sire type and a by-product based diet on performance and meat quality in growing-finishing pigs.

For many years, pig production has focused on maximizing performance by selecting for maximal muscle growth and feeding diets that allow the animals to express their genetic potential. However, it is unclear whether this selection for muscle deposition has affected the capacity of pigs to cope with by-product-based diets, which rely on fat as the primary energy source instead of starches and sugars. Therefore, an experiment was set up to investigate if different types of boars affect how their progeny cope with alternative ingredients in the diet, with a possible need for adapted breeding schemes. Two types of boars within the Piétrain sire line were used based on either a high or low estimated breeding value for daily feed intake (HFI: high feed intake, low feed intake). When their progeny reached 14 weeks of age, two dietary strategies were compared: a control (CON) vs a by-product-based diet high in fat and fiber (HFF). The CON diet was mainly based on cereals (corn, wheat, barley) and soybean meal. The HFF diet was formulated to contain the same net energy, CP and digestible amino acid levels without any cereals or soybean meal. In total 192 animals were included in the experiment (48 animals/type of boar/diet) and performance, digestibility, carcass and meat quality were compared. None of the parameters showed a significant interaction (P < 0.05) between the type of boar and diet, suggesting that shifting to diets that are less prone to feed-food competition is equally feasible in different types of pigs. Type of boar did affect performance, carcass quality and intramuscular fat content. HFI pigs showed higher daily feed intake (DFI) and daily gain (P < 0.001), with no significant difference in feed conversion ratio (P = 0.205), lower carcass quality (P < 0.001) and higher intramuscular fat content (P = 0.030). For both boar types, pigs fed the CON diet performed better, with a higher daily gain (P = 0.028), DFI (P = 0.011) and dressing yield (P = 0.009) and better digestibility (P < 0.001), but without differences in feed conversion ratio or meat quality. In conclusion, there was no indication that pigs differing in feed intake capacity cope differently with a high-fat, high-fiber diet based on by-products. Different types of pigs may cope well with diets that are less prone to feed-food competition.

Sarcopenia in end-stage liver disease and after liver transplantation.

Sarcopenia occurs in 30-70% of patients with end-stage liver disease and is associated with inferior pre- and post-liver transplant outcomes such as prolonged intubation times, long intensive care and hospitalization times, heightened risk of post-transplant infection, reduced health-related quality of life, and increased rates of mortality. The pathogenesis of sarcopenia is multifactorial and involves biochemical disturbances such as hyperammonemia, low serum concentrations of branched-chain amino acids (BCAAs) and low serum levels of testosterone, as well as chronic inflammation, inadequate nutritional status, and physical inactivity. Prompt recognition and accurate assessment of sarcopenia are critical and require imaging, dynamometry, and physical performance testing for the assessment of its subcomponents: muscle mass, muscle strength, and muscle function, respectively. Liver transplantation mostly fails to reverse sarcopenia in sarcopenic patients. In fact, some patients develop de novo sarcopenia after undergoing liver transplantation. The recommended treatment of sarcopenia is multimodal and includes a combination of exercise therapy and complementary nutritional interventions. Additionally, new pharmacological agents (e.g. myostatin inhibitors, testosterone supplements, and ammonia-lowering therapy) are under investigation in preclinical studies. Here, we present a narrative review of the definition, assessment, and management of sarcopenia in patients with end-stage liver disease prior to and after liver transplantation.

Effects of feeding guanidinoacetic acid on oxidative status and creatine metabolism in broilers subjected to chronic cyclic heat stress in the finisher phase.

Dietary guanidinoacetic acid (GAA) has been shown to affect creatine (Cr) metabolic pathways resulting in increased cellular Cr and hitherto broiler performances. Yet, the impact of dietary GAA on improving markers of oxidative status remains equivocal. A model of chronic cyclic heat stress, known to inflict oxidative stress, was employed to test the hypothesis that GAA could modify bird's oxidative status. A total of 720-day-old male Ross 308 broilers were allocated to 3 treatments: 0, 0.6 or 1.2 g/kg GAA was added to corn-SBM diets and fed for 39 d, with 12 replicates (20 birds each) per treatment. The chronic cyclic heat stress model (34°C with 50-60% RH for 7 h daily) was applied in the finisher phase (d 25-39). Samples from 1 bird per pen were taken on d 26 (acute heat stress) and d 39 (chronic heat stress). GAA and Cr in plasma were linearly increased by feeding GAA on either sampling day, illustrating efficient absorption and methylation, respectively. Energy metabolism in breast and heart muscle was greatly supported as visible by increased Cr and phosphocreatine: ATP, thus providing higher capacity for rapid ATP generation in cells. Glycogen stores in breast muscle were linearly elevated by incremental GAA, on d 26 only. More Cr seems to be directed to heart muscle as opposed to skeletal muscle during chronic heat stress as tissue Cr was higher in heart but lower in breast muscle on d 39 as opposed to d 26. The lipid peroxidation marker malondialdehyde, and the antioxidant enzymes superoxide dismutase and glutathione peroxidase showed no alterations by dietary GAA in plasma. Opposite to that, superoxide dismutase activity in breast muscle was linearly lowered when feeding GAA (trend on d 26, effect on d 39). Significant correlations between the assessed parameters and GAA inclusion were identified on d 26 and d 39 using principal component analysis. To conclude, beneficial performance in heat-stressed broilers by GAA is associated with enhanced muscle energy metabolism which indirectly may also support tolerance against oxidative stress.

Prevalence of Vertebral Fractures and Non-fracture Deformities in Healthy Adult Men: The Importance of Morphologic Criteria.

Vertebral fractures (VF) are common in older men but data on VF prevalence in young men is limited. The aim of this study was to describe the prevalence of VF and non-fracture vertebral deformities (VD) in healthy young to middle-aged men, and compare the characteristics of men with normal vertebrae, VF and VD. In this cross-sectional study, vertebral fracture assessment by dual-energy X-ray absorptiometry was performed in 650 men, aged 32 to 60 years (mean 46.2), from the population-based SIBLOS-SIBEX cohort. For VF and VD assessment, both the modified algorithm-based qualitative approach (morphologic criteria) to discriminate VF from VD and the semi-quantitative (morphometric) grading system of Genant (GSQ) were used. We found 48 (0.6%) fractured vertebrae, of which 15 were classified grade 1, 29 grade 2 and 4 grade 3 VF. There were 378 (4.7%) VD, of which 296 were scored grade 1, 82 grade 2 and none grade 3 VD. Twenty-six participants (4%) had VF, 15 had one and 11 had 2 or more VF. Two hundred and twenty-eight (35.1%) men had VD. Femoral neck, total hip and lumbar spine areal bone mineral density (aBMD) were lower in men with VF than in those with normal vertebrae or VD. Men with VD, in turn, had aBMD values similar to men with normal vertebrae. Our results suggest that -even in young healthy men-using the GSQ without taking qualitative aspects into account overestimates VF prevalence, confirming the importance of morphologic criteria to correctly diagnose and distinguish VF from VD.

Decrease of Salmonella and Escherichia coli O157:H7 counts during dry-aging of beef but potential growth of Listeria monocytogenes under certain dry-aging conditions.

Dry-aging of beef comprises the storage of carcasses and (sub)primal cuts at a low temperature and relative humidity for a prolonged period, aiming to increase the sensory quality of meat. Limited data are available on the survival and potential growth of pathogens on the surface of beef during dry-aging. Therefore, this study evaluates the changes in Salmonella, Shiga toxin-producing Escherichia coli O157:H7 and Listeria monocytogenes counts during dry-aging. A mixture of pathogenic strains was inoculated on the surface of beef loins, which were stored under four different process conditions (2 °C and 6 °C × relative humidity 75 and 85% during 42 days). Salmonella and E. coli O157:H7 counts significantly decreased during dry-aging. The daily reductions varied from -0.07 to -0.14 log10 CFU and from -0.09 to -0.14 log10 CFU, respectively, depending on the loin, matrix and condition. The reduction of L. monocytogenes was slower, with a maximum of -0.07 log10 CFU/day. L. monocytogenes counts increased with 1.0 log10 CFU on the lean meat of one loin with pH > 6.0 at the end of dry-aging, indicating that this pathogen can potentially grow under certain dry-aging conditions.

Influence of aging time, temperature and relative humidity on the sensory quality of dry-aged Belgian Blue beef.

The objective of this study was to evaluate the effect of aging period (0, 3, 6 or 9 weeks), aging temperature (2 versus 6 °C at 75% relative humidity, experiment 1) and relative humidity (70 versus 90% at 2 °C, experiment 2) on the sensory traits, oxidative stability and proteolysis of Belgian Blue beef. For each experiment, eight loins (M. longissimus thoracis et lumborum) from four animals (left and right side) were assigned to one of the two treatments (n = 4). Results showed no further tenderization after three weeks of aging, whereas metmyoglobin formation and lipid oxidation increased until nine weeks of aging (P < 0.05). During the nine weeks of aging, atypical flavor, odor and flavor intensity was affected (P < 0.05). This was accompanied by an increase of small peptides and other nitrogenous compounds. Aging temperature and relative humidity had only a very limited effect on the quality traits.

In vitro stability and ex vivo absorption of thymol monoglucosides in the porcine gut.

Thymol α-D-glucopyranoside (TαG) and thymol ß-D-glucopyranoside (TßG) are believed to have different kinetic behaviours in the porcine gut than its parent aglycon thymol. However, recently, it was shown that concentrations of both glucosides decreased rapidly in the stomach and proximal small intestine following oral supplementation to piglets as did thymol. Yet, the stability of thymol glucosides in gut contents and their absorption route remains obscure. Therefore, a series of in vitro incubations were performed, simulating the impact of pH, digestive enzymes, bacterial activity and mucosal extracts on stability of these glucosides. Their absorption mechanisms were investigated using the Ussing chamber model in the presence or the absence of inhibitors of sodium-dependent glucose linked transporter 1 and lactase phlorizin hydrolase. Both glucosides remained intact at physiological pH levels in the presence of digestive enzymes. Recoveries from TαG and TßG were below 90% when incubated with small intestinal homogenates from the distal jejunum or from all sampled sites, respectively. However, no aglycon could be detected in these samples. Bacterial inoculum of the small intestine, on the other hand, hydrolysed TßG quickly with up to 44% of free aglycon appearing. TαG proved more resistant to porcine gastro-intestinal bacterial glucosidases with only trace amounts (<1%) of free thymol at the end of the incubations. Electrophysiological measurements in Ussing chambers did not suggest active transport of the glucosides. Mucosal TαG and TßG concentrations were unchanged between start and end of the absorption measurements. Additionally, no TαG and only a very limited amount of TßG were retrieved from the serosal side. Tissue associated concentrations, although marginal (<1% of luminal concentration), were mainly as intact glucoside or as aglycon for TαG and TßG, respectively. Addition of both inhibitors significantly increased the amount of intact glucosides retrieved from the mucosal tissues as compared to controls. In conclusion, bacterial hydrolysis was identified as the most important source of TßG loss, whereas TαG seemed less prone to degradation or absorption in these in vitro and ex vivo models.

Gastrostomy for infants with severe epidermolysis bullosa simplex in neonatal intensive care.

INTRODUCTION: Severe epidermolysis bullosa simplex (EBS sev) is a rare genodermatosis characterized by congenital generalized blistering and mucosal involvement. Increased needs and decreased intake quickly lead to nutritional imbalance. Enteral nutrition support is proposed, but classical nasogastric tubes are not well tolerated in these patients and gastrostomy is preferred. OBJECTIVE AND METHODS: To report the experience with EBS sev in neonatal units of French reference centers for gastrostomy. In this retrospective multicentric study, we included all patients with EBS sev who had gastrostomy placement before age 9 months during neonatal care hospitalization. RESULTS: Nine infants (5 males/4 females) with severe skin and mucosal involvement were included. A gastrostomy was decided, at an early age (mean 3.7 months, range 1.4 to 8 months) in infants with mean weight 4426 g (range 3500 to 6000 g). Techniques used were endoscopy with the pull technique for 5 infants and surgery under general anesthesia for 4. Main complications were local but resolved after treatment. All infants gained weight after gastrostomy. The mean withdrawal time (n = 7) for the gastrostomy was 35.8 months (range 10.5 months to 6.5 years). Seven children had persistent oral disorders. CONCLUSIONS: Gastrostomy in infants with EBS sev can be necessary in neonatal intensive care units. Both surgical and endoscopic pull techniques seem efficient, with good tolerance.

Acute preexercise supplementation of combined carnosine and anserine enhances initial maximal power of Wingate tests in humans.

Classic in vitro experiments (Severin's phenomenon) demonstrated that acute carnosine supplementation may potentiate muscle contractility. However, upon oral ingestion, carnosine is readily degraded in human plasma by the highly active serum carnosinase-1 (CN1). We developed a novel strategy to circumvent CN1 by preexercise ingestion of combined carnosine (CARN) and anserine (ANS), the methylated analog with similar biochemical properties but more resistant to CN1. First, in vitro hydrolysis was tested by adding carnosine and anserine to human plasma, alone or in combination. Second, five subjects were supplemented with 25 mg/kg anserine or 25 mg/kg of each anserine and carnosine to test in vivo bioavailability. Third, two double-blind, placebo-controlled, crossover studies investigated the effect of preexercise ANS + CARN (20 mg/kg body wt of each) supplementation on performance during a single all-out Wingate test following 6-min high-intensity cycling (study A) or three repeated Wingate tests (study B). In vitro experiments demonstrated slower degradation of anserine versus carnosine, which was further slowed by simultaneously adding carnosine. In vivo bioavailability of plasma anserine was more prominent [2.5-fold increased area under the curve (AUC)] when ANS + CARN versus ANS was ingested. Study A showed significantly higher (+6% ± 11%; P = 0.04) power in the first 5 s of the Wingate test following ANS + CARN (12.8 ± 2.4 W/kg) versus placebo (12.1 ± 2.2 W/kg). Study B demonstrated increased peak power (+3%) throughout three consecutive Wingate tests (ANS + CARN 10.5 ± 0.6 W/kg vs. placebo 10.2 ± 9.9 W/kg). These experiments reveal a novel acute nutritional method to effectively raise plasma anserine and carnosine by high-dose combined supplementation. This approach led to improved initial cycling power, revealing a new nutritional strategy to increase exercise performance.NEW & NOTEWORTHY Current results reveal that carnosine and anserine competitively bind to the highly active carnosinase enzyme in human plasma. Acute combined carnosine and anserine supplementation is therefore described as novel strategy to raise plasma anserine and carnosine. We report that indices of maximal exercise/muscle power during the initial stage of a Wingate test were significantly improved by preexercise 20-25mg/kg body wt anserine and carnosine supplementation, pointing toward a novel acute nutritional strategy to improve high-intensity exercise performance.

Effect of terminal sire line and timing second vaccination on effectiveness of immunocastration, performance, and carcass and meat quality.

Performance (from 10 weeks until slaughter), carcass and meat quality, and effectiveness of immunocastration was compared in crossbred offspring of stress positive (BP+) and negative (BP-) Belgian Piétrain and Canadian Duroc (CD) given the second vaccination of Improvac® at different times (4, 6, 8 weeks before slaughter). CD offspring had a significantly higher daily gain (DG) and feed intake (DFI), and lower predicted lean meat percentage (LMP) and dressing yield compared to BP+ and BP-, while feed conversion ratio (FCR) did not differ. CD offspring had significantly lower drip loss and higher pHi, intramuscular fat content than BP+ and BP- (except for pHi). No significant effect of vaccination time on DG nor FCR was observed. Predicted LMP tended to increase as time-post injection decreased, while meat quality was minor affected. Earlier vaccination had no effect on the effectiveness of immunocastration based on testosterone and GnRH-binding.

Quality characteristics of fresh loin and cooked ham muscles as affected by genetic background of commercial pigs.

This study was performed to investigate meat quality traits of loin and ham of commercial pigs as affected by genetic differences in carcass and growth traits of the parent lines. Three hybrid sow lines were crossbred with two types of Belgian Piétrain with different breeding goals (BPgrowth and BPcarcass emphasizing daily growth and carcass conformation, respectively). Pig live performance and carcass quality of 270 offspring were measured, and meat quality of the loin and (cooked) ham was evaluated on 216 animals. Despite the differences in pig live performance and carcass quality for sow line, little effect on meat quality was observed. Only a lower (p < 0.05) intramuscular fat content of ham and a tendency (p < 0.1) toward lower cooking yield was observed in offspring of the sow line with the highest versus the lowest carcass lean content. Loin traits were only weakly associated with fresh and cooked ham quality traits.

Guanidinoacetic acid supplementation improves feed conversion in broilers subjected to heat stress associated with muscle creatine loading and arginine sparing.

It was hypothesized that dietary guanidinoacetic acid (GAA), the precursor of creatine (Cr), would be beneficial to heat-stressed finisher broilers owing to improved cellular energy status and arginine sparing effects. A total of 720 one-day-old male Ross 308 broilers were allocated to 3 treatments, 0 (control), 0.6, or 1.2 g/kg of GAA added to complete corn-soybean meal diets, and were fed for 39 D, with 12 replicates (20 birds each) per treatment. A chronic cyclic heat stress model (at a temperature of 34°C and 50 to 60% relative humidity for 7 h daily) was applied in the finisher phase (day 25-39). Samples were taken on day 26 and 39 to determine thrombocyte, white blood cell, corticosterone, protein and amino acid levels in blood and Cr, phosphocreatine (PCr), and adenosine triphosphate levels in the breast muscle. Meat quality was assessed on day 40 after overnight fasting. Guanidinoacetic acid at a dose of 1.2 g/kg decreased feed-to-gain ratio compared with the control in the grower phase (1.32 vs. 1.35, respectively; P <0.05). In the finisher period, the supplementation of 1.2 g/kg of GAA reduced feed intake compared with the control (-3.3%, P <0.05), whereas both GAA supplementation levels improved feed efficiency markedly (1.76, 1.66, and 1.67 for 0 [control], 0.6, and 1.2 g/kg of GAA, respectively, P <0.05). Mortality outcomes highlight that GAA feeding improved survival during heat stress, supported by lower panting frequency (linear effect, P <0.05). Plasma arginine was higher with increase in dietary GAA concentration on day 26 (+18.3 and + 30.8% for 0.6 and 1.2 g/kg of GAA, respectively; P <0.05). This suggests enhanced availability of arginine for other metabolic purposes than de novo GAA formation. In the breast muscle, PCr (day 39, P <0.05), free Cr (day 39, P <0.05), total Cr (both days, P <0.05), and PCr-to-adenosine triphosphate ratio (day 39, P <0.05) levels were increased with higher GAA content in diet. Guanidinoacetic acid supplementation improved feed conversion and survival during chronic cyclic heat stress, which may be associated with enhanced breast muscle energy status and arginine sparing effect.

Performance and carcass, loin and ham quality in crossbreds from three terminal sire lines.

This study was designed to compare performance, carcass and meat quality of crossbred of a hybrid sow x three sire lines, i.e. stress positive Belgian Piétrain (BP), stress negative French Piétrain (FP) and Canadian Duroc (CD). BP offspring had a significantly higher carcass yield (p < .001) and lean meat content (p < .001) in comparison with FP, which was higher than CD. BP offspring had significantly lower pH (p < .05), water-holding capacity (WHC) (p < .001) and intramuscular fat (IMF) (p < .001) content in the loin compared to FP and CD, but these meat quality parameters, with the exception of pH, were superior for CD as compared to FP. In accordance with loin quality, pHi, pHu, WHC and IMF of BP were significantly lower (p < .05) compared to CD in the fresh and cooked ham. Most often, FP offspring could not be differentiated from the other offspring, with the exception of cooking loss of the cooked ham. Trained and consumer taste panels resulted in no significant differences (p > .1) in sensory attributes, however, consumers preffered CD based on ranking.

Effects of methionine and guanidinoacetic acid supplementation on performance and energy metabolites in breast muscle of male broiler chickens fed corn-soybean diets.

1. Guanidinoacetic acid (GAA) is the single endogenous precursor of creatine, which plays a critical role in energy homeostasis of cells. Since GAA is endogenously converted to creatine by methylation, it was hypothesised that the effects of dietary GAA supplementation might determine the methionine (Met) availability in corn-soybean based diets. 2. A total of 540, one-day-old male Ross 308 broilers were allocated to nine dietary treatments with six replicates (10 birds each) in a 3 × 3 factorial arrangement with three graded levels of supplementary Met (+0.4 g/kg per level), whilst cystine was equal across groups, resulting in a low, medium and high level of total sulphur amino acids, and with three levels of GAA (0, 0.6 and 1.2 g/kg). Birds were fed for 42 days. 3. Increasing levels of supplemental Met enhanced performance indices in all rearing periods, although there was no effect on feed conversion ratio in the grower or feed intake in the finisher periods. Final body weight was 8.8% and 14.6% higher in the birds fed medium and high Met diets, respectively, compared to the low Met level. Relative breast weight and protein content in muscle on d 25 linearly increased with higher levels of Met. At low and high Met levels, growth in the finisher phase was negatively affected by supplementing GAA at 1.2 g/kg. It was suggested that disturbances in methylation homeostasis and/or changes in Arg metabolism might explain these findings. At the end of the grower phase, muscle creatine content was higher when feeding GAA at 0.6 and 1.2 g/kg (4464 and 4472, respectively, vs. 4054 mg/kg fresh muscle in the control group). 4. The effects of dietary GAA supplementation were influenced by the dietary Met level only in the finisher period, which indicates the need for proper sulphur amino acid formulation in diets when feeding GAA.

Impact of a dietary challenge with peroxidized oil on the glutathione redox status and integrity of the small intestine in weaned piglets.

Glutathione (GSH) is considered to play an important role in maintaining the integrity of the small intestine. In piglets, altered mucosal GSH levels might therefore be involved in weaning-induced changes of the small intestinal morphology and barrier function. To test this hypothesis, we aimed to challenge the mucosal GSH redox status during the first 28 days after weaning, by feeding diets containing 5% fresh linseed oil (CON), or 2.5% (OF1) or 5% (OF2) peroxidized linseed oil (peroxide value 225 mEq O2/kg oil) and exploring the effects on gut integrity. Piglets were pair-fed and had a total daily feed allowance of 32 g/kg BW. A fourth treatment included animals that were fed the control diet ad libitum (ACON). Animals were sampled at days 5 and 28 post-weaning. The malondialdehyde (MDA) concentration and GSH redox status (GSH/GSSG Eh) were determined in blood, liver and small intestinal mucosa. Histomorphology of the duodenal and jejunal mucosa was determined, and Ussing chambers were used to assess fluorescein isothiocyanate dextran (FD4) and horseradish peroxidase (HRP) fluxes across the mucosa. Results show that peroxidized linseed oil imposed an oxidative challenge at day 28, but not at day 5 post-weaning. At day 28, increasing levels of dietary peroxides to pair-fed pigs linearly increased MDA levels in duodenal and jejunal mucosa. Moreover, FD4 fluxes were significantly increased in OF1 (+75%) and OF2 (+64%) in the duodenum, and HRP fluxes tended (P=0.099) to show similar differences, as compared to CON. This co-occurred with a significant 11 mV increase of the hepatic GSH/GSSG Eh, potentiated by a significantly increased GSH peroxidase activity for treatments OF1 (+47%) and OF2 (+63%) in liver as compared to CON. Furthermore; duodenal HRP flux significantly correlated with the hepatic glutathione disulphide (GSSG) level (r=0.650), as also observed in the jejunum for hepatic GSSG (r=0.627) and GSH/GSSG Eh (r=0.547). The jejunal permeability was not affected, but FD4 and HRP fluxes significantly correlated with the local GSH (r=0.619; r=0.733) and GSSG (r=0.635; r=0586) levels. Small intestinal histomorphology was not affected by dietary lipid peroxides, nor were there any correlations found with the GSH redox system. To conclude, under oxidative stress conditions, jejunal barrier function is related to the local and hepatic GSH redox system. It is suggested that the hepatic GSH system participates in the elimination of luminal peroxides, and thereby impacts on duodenal barrier function.

An accelerated, clinical-grade protocol to generate high yields of type 1-polarizing messenger RNA-loaded dendritic cells for cancer vaccination.

BACKGROUND: Many efforts have been devoted to improve the performance of dendritic cell (DC)-based cancer vaccines. Ideally, a DC vaccine should induce robust type 1-polarized T-cell responses and efficiently expand antigen (Ag)-specific cytotoxic T-cells, while being applicable regardless of patient human leukocyte antigen (HLA) type. Production time should be short, while maximally being good manufacturing practice (GMP)-compliant. We developed a method that caters to all of these demands and demonstrated the superiority of the resulting product compared with DCs generated using a well-established "classical" protocol. METHODS: Immunomagnetically purified monocytes were cultured in a closed system for 3 days in GMP-compliant serum-free medium and cytokines, and matured for 24 h using monophosphoryl lipid A (MPLA)+ interferon-gamma (IFN-γ). Mature DCs were electroporated with messenger RNA (mRNA) encoding full-length antigen and cryopreserved. "Classical" DCs were cultured for 8 days in flasks, with one round of medium and cytokine supplementation, and matured with tumor necrosis factor alpha (TNF-α) + prostaglandin E2 (PGE2) during the last 2 days. RESULTS: Four-day MPLA/IFN-γ-matured DCs were superior to 8-day TNF-α/PGE2-matured DCs in terms of yield, co-stimulatory/co-inhibitory molecule expression, resilience to electroporation and cryopreservation and type 1-polarizing cytokine and chemokine release after cell thawing. Electroporated and cryopreserved DCs according to our protocol efficiently present epitopes from tumor antigen-encoding mRNA, inducing a strong expansion of antigen-specific CD8+ T-cells with full cytolytic capacity. CONCLUSION: We demonstrate using a GMP-compliant culture protocol the feasibility of generating high yields of mature DCs in a short time, with a superior immunogenic profile compared with 8-day TNF-α/PGE2-matured DCs, and capable of inducing vigorous cytotoxic T-cell responses to antigen from electroporated mRNA. This method is now being applied in our clinical trial program.

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density.

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets. (2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial. (3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle). (4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.

The potential of herbs and spices to reduce lipid oxidation during heating and gastrointestinal digestion of a beef product.

Here we tested the potential of ten culinary herbs (basil, oregano, rosemary and thyme) and spices (black pepper, cayenne pepper, cumin, curcuma, garlic and sweet paprika) to limit oxidation during cooking of a high-fat beef product, and during its subsequent in vitro gastrointestinal digestion. The herbs and spices were added separately at 0.5% and 1% (w/w), either during meat processing before heating of the product or after heating of the meat as a seasoning. Lipid oxidation was evaluated by malondialdehyde, 4-hydroxy-2-nonenal and hexanal quantification in the meat products and gastrointestinal digests. In general, adding the herbs and spices before heating of the meat was more effective at limiting oxidation during digestion, compared to adding these compounds after heating. Doses of 1% were more effective than 0.5%. The four herbs and curcuma displayed antioxidant activity in all treatments and were more effective in limiting oxidation than the other culinary compounds, resulting in negligible concentrations of 4-hydroxy-2-nonenal and hexanal in the meat and digests. Black pepper, cayenne pepper and cumin moderately reduced or had no effect on oxidation during digestion when added following heating of the meat, but had a more pronounced antioxidant effect when added before heating. Sweet paprika and garlic exerted some pro-oxidant effects during cooking. During digestion, sweet paprika had an antioxidant effect, but garlic did not. The extent to which the herbs and spices limited oxidation, showed a strong correlation with their phenolic content, and lower correlations with α-tocopherol and chlorophyll.

Association between overweight/obesity and periodontal disease in children and adolescents: a systematic review and meta-analysis.

AIM: To provide a systematic review and meta-analyses investigating the association between overweight/obesity as defined by Body Mass Index (BMI) and periodontal disease in terms of clinical periodontal outcomes. MATERIALS AND METHODS: A systematic search of the literature was conducted by two authors (SR and SD) independently in the Cochrane Library, PubMed, Web of Science (ISI), Scopus, Scielo, Lilacs and System for Information on Grey Literature in Europe (SIGLE) for full articles published until September 2015. Studies analysing the association between overweight/obesity as defined by Body Mass Index (BMI) and periodontal disease in children and/or adolescents (age ≤18 years) were included. The Gwets AC1 inter-rater reliability coefficient for screening data was calculated using Agreestat 2011.1. Meta-analyses were carried out by using RStudio version 0.97.551-©2009-2012 RStudio, Inc. software. RESULTS: A total of 769 titles and abstracts were screened and 12 articles met the inclusion criteria for the systematic review while only 7 were selected for meta-analyses. The Gwets AC1 inter-rater reliability coefficient for screening data was excellent (0.98; CI 0.98-0.99). A positive association between overweight/obesity and a number of periodontal diseases was seen. For the association between prevalent periodontal disease and obesity in children, the overall fixed-effects OR and 95% CI was 1.46 (1.20-1.77) with a χ2 statistic for heterogeneity (Q) of 33.4 with 6 degrees of freedom (P < 0.005). CONCLUSION: The available evidence suggests a significantly positive association between periodontal disease and obesity in children. Paediatric dentists should be aware of periodontal alterations as a potential hazard associated with obesity.