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Exposure to high ambient temperature has been shown to impair growth performance and to cause oxidative stress in broilers. This study investigated the hypothesis that supplementation with methionine (Met) as DL-Met (DLM) more than the National Research Council recommendations improves growth performance and alleviates oxidative stress in broilers exposed to high ambient temperature. One-day-old male Cobb-500 broilers (n = 68) were allotted to 4 groups and phase-fed 3 basal diets during days 1 to 10, 11 to 21, and 22 to 35. One group was kept under thermoneutral temperature conditions and received the basal diets with Met + cysteine (Cys) concentrations according to recommendations of NRC. The other 3 groups were kept in a room with an increased ambient temperature from week 3 to 5 and were fed either the basal diet or the basal diets supplemented with 2 levels of DLM in which Met + Cys concentrations exceeded NRC recommendations by around 20% (group DLM1) and 40% (group DLM2), respectively. As expected, the broilers exposed to high ambient temperature showed a lower feed intake, lower body weight gains, a higher feed:gain ratio, and biochemical indications of oxidative stress in comparison to broilers kept under thermoneutral temperature conditions. Supplementation of DLM did not improve the growth performance in broilers exposed to high ambient temperature. However, the broilers supplemented with DLM had increased concentrations of glutathione in liver and breast muscle (groups DLM1 and DLM2), increased concentrations of tocopherols in the liver (group DLM2), and reduced concentrations of 7α-hydroxycholesterol and 7-ketocholesterol in heat-processed thigh muscle (groups DLM1 and DLM2) in comparison to the control group exposed to high ambient temperature. Concentrations of thiobarbituric acid-reactive substances and vitamin C in plasma, liver, and muscle were not different between the 3 groups exposed to heat stress. Nevertheless, the study shows that supplementation of DLM in slight excess of the Met concentration required for maximum growth performance improved the antioxidant status in tissues and reduced the susceptibility of muscle toward oxidation in heat-stressed broilers.

This study aimed to investigate the effects of a short-term (36 h) fasting period combined with an acute bout of exercise on markers of immune function and inflammation in healthy human subjects. Fourteen moderately trained male subjects (aged 19–39 yr) participated in a 36-h fasting trial (FA-T), followed by an acute bout of moderate exercise (60% V̇o2max). After 1 wk, the same subjects, as their own control, participated in a nonfasting trial (NFA-T) in which they performed an exercise trial of the same duration and intensity. Blood samples were taken before, immediately after, and 1 h after each exercise bout and analyzed for several immunological and metabolic markers. At baseline, fasting subjects showed lower levels of T cell apoptosis, lymphocyte-proliferative responses, IL-6, monocyte chemoattractant protein-1 (MCP-1), insulin, and leptin ( P < 0.05) as well as higher levels of neutrophil oxidative burst and thiobarbituric acid reactive substances (TBARS) than those in the NFA-T ( P < 0.05). After the exercise protocol, fasted subjects revealed higher T cell apoptosis, neutrophil oxidative burst, TBARS, TNFα, and MCP-1 levels as well as lower levels of lymphocyte-proliferative response, IL-6, insulin, and leptin than those in the NFA-T ( P < 0.05). Short-term fasting aggravates perturbations in markers of immune function, and inflammation was induced by an acute moderate-intensity exercise protocol.

Insect biomass obtained from large-scale mass-rearing of insect larvae has gained considerable attention in recent years as an alternative and sustainable source of food and feed. A byproduct from mass-rearing of insect larvae is the shed cuticles - the most external components of insects which are a relevant source of the polysaccharide chitin. While it has been shown that chitin modulates the gut microbiota and ameliorates lipid metabolic disorders in obese rodent models, feeding studies dealing with isolated insects' cuticles are completely lacking. Thus, the present study tested the hypothesis that dietary insects' cuticles modulate the gut microbiome and improve hepatic lipid metabolism in obese Zucker rats. To test this hypothesis, three groups of obese Zucker rats were fed a nutrient-adequate, semisynthetic basal diet which was supplemented with either 0% (group O), 1.5% (group O1.5) or 3.0% (group O3.0)

Insect meal (IM) produced from edible insects, such as

This study investigated the hypothesis that dietary supplementation of leucine (Leu) above actual recommendations activates protein synthesis and inhibits protein degradation pathways on the molecular level and supports higher muscle growth in broilers. Day-old male Cobb-500 broilers (n = 180) were allotted to 3 groups and phase-fed 3 different corn-wheat-soybean meal-based basal diets during periods 1 to 10, 11 to 21, and 22 to 35 D. The control group (L0) received the basal diet which met the broiler's requirements of nutrients and amino acids for maintenance and growth. Groups L1 and L2 received basal diets supplemented with Leu to exceed recommendations by 35 and 60%, respectively, and isoleucine (Ile) and valine (Val) were supplemented to keep Leu: Ile and Leu: Val ratios fixed. Samples of liver and breast muscle and pancreas were collected on days 10, 21, and 35. The gene expression and abundance of total and phosphorylated proteins involved in the mammalian target of rapamycin pathway of protein synthesis, in the ubiquitin-proteasome pathway and autophagy-lysosomal pathway of protein degradation, in the general control nonderepressible 2/eukaryotic translation initiation factor 2A pathway involved in the inhibition of protein synthesis, and in the myostatin-Smad2/3 pathway involved in myogenesis were evaluated in the muscle, as well as expression of genes involved in the growth hormone axis. Growth performance, feed intake, the feed conversion ratio, and carcass weights did not differ between the 3 groups (P > 0.05). Plasma concentrations of Leu, Ile, and Val and of their keto acids, and the activity of the branched-chain α-keto acid dehydrogenase in the pancreas increased dose dependently with increasing dietary Leu concentrations. In the breast muscle, relative mRNA abundances of genes and phosphorylation of selected proteins involved in all investigated pathways were largely uninfluenced by dietary Leu supplementation (P > 0.05). In summary, these data indicate that excess dietary Leu concentrations do not influence protein synthesis or degradation pathways, and subsequently do not increase muscle growth in broilers at fixed ratios to Ile and Val.

Intensive milk feeding and butyrate supplementation in calves stimulate body growth and affect gastrointestinal development. The aim of the present study was to investigate the synergistic effects of ad libitum milk replacer (MR) feeding and butyrate supplementation of MR on rumen and small intestinal growth and on gene expression in the small intestine related to growth and energy metabolism at weaning. Male Holstein calves (n = 32) received colostrum from birth to d 3 of age and MR either ad libitum (Adl) or restrictively (Res; 6 L of MR/d; 12.5% solids) with (AdlB+, ResB+) or without (AdlB−, ResB−) 0.24% butyrate from d 4 until wk 8 of age. From wk 9 to 10, all calves were weaned and were fed 2 L/d until the end of the trial. Concentrate, hay, and water were freely available. At d 80, calves were slaughtered, volatile fatty acids were measured in rumen fluid, and rumen and small intestine samples were taken for histomorphometric measurements. The expression of mRNA associated with the local insulin-like growth factor (IGF) system and glucose metabolism as well as lactase and maltase activities were measured in the intestinal mucosa. The small intestine was 3 m longer in Adl than in Res. In the atrium ruminis, papilla width was greater in Res than in Adl. Villus circumference, cut surface, and height in the duodenum, proximal jejunum, and ileum were greater in Adl than in Res and in the proximal, mid, and distal jejunum and ileum were greater in calves treated with butyrate. Crypt depth in the duodenum and proximal jejunum was greater in Adl than in Res and in the ileum was smaller in calves treated with butyrate. The villus height:crypt depth ratio was greatest in AdlB+ calves. In the proximal and mid jejunum, IGF1 mRNA abundance was lower in calves treated with butyrate. In the proximal jejunum, INSR mRNA abundance was greater in Res than in Adl. The abundance of PCK2 mRNA was greater in Res than in Adl in the duodenum and was greatest in ResB− in the mid jejunum. Lactase activity tended to be greater in Res than in Adl and after butyrate treatment in the proximal jejunum. The results indicated an elevated growth of the small intestinal mucosa at weaning due to intensive milk feeding and butyrate supplementation, and the local IGF system was involved in intestinal growth regulation. Rumen development was not affected by butyrate supplementation of MR and was slightly delayed due to ad libitum MR feeding.

The present study tested the hypothesis that the liver lipid-lowering effect of insect meal (IM) is caused by its low methionine concentration. A total of fifty, male obese Zucker rats were randomly assigned to five groups of 10 rats each (casein (C), IM, IM + Met, IM + Cys, and IM + EAA). While group C received a diet with casein, the IM-fed groups received a diet with IM as the protein source. In groups IM + Met, IM + Cys and IM + EAA, the diets were additionally supplemented with methionine, cysteine and essential amino acids (EAA), respectively. Hepatic concentrations of triacylglycerols and cholesterol, and hepatic mRNA levels and activities of lipogenic and cholesterogenic enzymes were markedly lower in the IM-fed groups than in group C (p &lt, 0.05). All of these parameters either did not differ across the IM-fed groups or were only slightly higher in groups IM + Met, IM + Cys and IM+EAA than in the group IM. In conclusion, the results indicate that a difference in the amino acid composition between IM and casein, a low concentration of methionine in IM and a reduced cysteine synthesis secondary to a decreased methionine availability resulting from feeding IM are not causative for the lipid-lowering effect of IM.

This study investigated the hypothesis that supplementation of methionine (Met) to broiler diets increases muscle growth due to regulation of molecular pathways related to protein synthesis and degradation depending on the Met source. Day-old male Cobb-500 broilers (n = 240) were phase-fed three different wheat-soya bean meal-based basal diets during days 1-10, 11-21 and 22-35. Basal diets (Met- group, Met + Cys concentration 15% below NRC recommendations) were supplemented with 0.10% or 0.40% Met either as DL-Met (DLM) or DL-2-hydroxy-4-(methylthio) butanoic acid (DL-HMTBA) (equimolar comparison). Breast muscle weights were lower in the Met- group compared to all Met-supplemented groups and were lower in broilers supplemented with 0.10% of DL-HMTBA compared to the other groups fed Met-supplemented diets. However, the expression of genes or relative phosphorylation and thus activation state of proteins involved in the somatotropic axis, the mammalian target of rapamycin (mTOR) pathway of protein synthesis, the ubiquitin-proteasome pathway (UPP) and autophagy-lysosomal pathway of protein degradation, the GCN2/eIF2a pathway involved in the inhibition of protein synthesis and in the myostatin-Smad2/3 pathway involved in myogenesis were not affected by Met source. Feeding diets with suboptimum Met + Cys concentrations, however, decreased expression of GHR and IGF1 in liver and muscle and increased that of MURF1 involved in the UPP in the broiler's muscle at day 10 and 21, while that of FOXO and atrogin-1 and FOXO phosphorylation remained unaffected. Additionally, suboptimum dietary Met concentrations increased expression of the autophagy-related genes ATG5 and BECN1 at day 35. Met supplementation neither affected gene expression nor phosphorylation of proteins involved in the GNC2/eIF2a and mTOR pathways. These data indicate that protein synthesis was not affected on the molecular level, while protein degradation was marginally affected by dietary Met dosage.

PURPOSE To evaluate the sinus membrane perforation and implant survival rate after crestal minimally invasive sinus floor augmentation using hydraulic pressure and vibrations. MATERIALS AND METHODS In this retrospective single cohort study, all patients who underwent minimally invasive sinus floor augmentation between 2007 and 2015 using hydraulic pressure and vibrations were included. The sinus membrane is elevated by physiologic saline at 1.5 bar. The fluid is then set into vibration to further separate the sinus membrane from the bony floor. The endpoints were sinus membrane perforation and the survival rate of implants. RESULTS The hydraulic pressure and vibration technique was applied in 156 patients. Seven patients with perforations of the sinus membrane were treated with the lateral window approach and excluded from the follow-up analysis. In the remaining 149 patients, 184 crestal sinus floor augmentations were performed and 184 implants were placed. In 10 of these 184 cases, a perforation was suspected in the postoperative computed tomography (CT) scan. In total, the perforation rate was 8.9% (17/191). Nineteen implants were lost during the follow-up period ranging from 0.2 to 8.4 years with a median of 2.3 years. The cumulative implant survival rates after 1, 3, and 5 years were 94.4%, 87.7%, and 87.7%, respectively. No severe perioperative complications were noted. CONCLUSION The hydraulic pressure and vibration technique allows a minimally invasive crestal sinus augmentation with a perforation rate less than 10% and implant survival rates of approximately 90%.

This study hypothesized that plasma and tissue antioxidant status of broilers is positively influenced when dietary Met concentrations exceed, and negatively when they go below NRC recommendations. In addition, different Met sources are hypothesized to affect the antioxidant defence system differently. Day-old male Cobb-500 broilers (n = 336) were allotted to seven groups and phase-fed three wheat-soya bean meal-based basal diets during days 1-10, 11-21 and 22-35. The basal diets (Met- group, Met + Cys concentration 15% below NRC recommendations) were supplemented with 0.10%, 0.25% or 0.40% Met either as DL-Met (DLM) or DL-2-hydroxy-4-(methylthio) butanoic acid (DL-HMTBA) (equimolar comparison). Growth performance and carcass weights were lower in the Met- group compared to the groups whose diets met or exceeded Met requirements. The antioxidant defence system was not influenced by the Met source. However, in the liver, concentrations of glutathione increased with increasing dietary Met concentrations. Tocopherol concentrations in the liver at days 10 and 21 were lower in the Met- group than in the groups supplemented with Met. However, liver concentrations of thiobarbituric acid reactive substances (TBA-RS) and protein carbonyls (PC) were largely not influenced by dietary Met concentration. Plasma tocopherol concentrations at day 35 were lower, and those of TBA-RS and PC at day 35 were higher in Met- group than in the groups fed the Met-supplemented diets. In jejunum, but not in liver, relative mRNA abundances and activities of superoxide dismutase, catalase and glutathione peroxidase were higher in the Met- group than in the groups fed Met-supplemented diets. These data indicate that suboptimum supply of Met results in decreased antioxidant concentrations in plasma and body tissues, and increases oxidative stress in the jejunum mucosa. However, supplementation of Met in excess of the requirements (based on NRC) compared to diets adequate in Met + Cys did not influence the antioxidant defence system.

Long-term cigarette smoking induces inflammatory processes in the pulmonary system that are suggested to “spill over” into systemic inflammation. Regular exercise has been shown to have anti-inflammatory properties. The aim of the study was to investigate the effects of therapeutic exercise on inflammation and muscle wasting in smoke-exposed mice. C57BL/6J mice ( n = 30) were separated into three groups to receive either 1) no specific treatment (control group), 2) 8-mo exposure to cigarette smoke [smoke-exposed (SE) group], or 3) 8 mo of cigarette smoke combined with exercise training during the last 2 mo (SEex group). The inflammatory status was analyzed by quantifying levels of various plasma proteins using multiplex ELISA and detection of lymphocyte surface markers by flow cytometry. Muscle tissue was analyzed by histological techniques and measurements of RNA/protein expression. SE led to decreased maximal O2 uptake (V̇o2max) and maximal running speed ( Vmax), which was reversed by exercise ( P < 0.05). Expression of ICAM-1, VCAM-1, and CD62L on T cells increased and was reversed by exercise ( P < 0.05). Similarly, SE induced an increase of various inflammatory cytokines, which were downregulated by exercise. In muscle, exercise improved the structure, oxidative capacity, and metabolism by reducing ubiquitin proteasome system activation, stimulating insulin-like growth factor 1 expression, and the SE-induced inhibition of mammalian target of rapamycin signaling pathway ( P < 0.05). Exercise training reverses smoke-induced decline in exercise capacity, systemic inflammation, and muscle wasting by addressing immune-regulating, anabolic, and metabolic pathways.

Purpose The aim of the current study was to investigate the effects of the probiotic Escherichia coli strain Nissle 1917 (EcN) on the exercise-induced disruption of gastrointestinal (GI) integrity and the associated release of damage and inflammatory markers. Methods After a pre-performance test, 19 untrained subjects (aged 18–35 years) passed two identical exhaustive treadmill exercise tests in an intensity corresponding to 60–80% VO2max in a test–retest design. The exercise tests were separated by a time period of 4 weeks. During this period, all subjects ingested 5 ml of an EcN suspension daily. Serum samples were taken before, immediately following and 3 h after both exercise tests. They were analyzed for indicators of GI integrity (zonulin; claudin-3; LPS), various damage and redox markers (I-FABP, GOT; GPT; TBARS) and inflammatory parameters (hsCRP; leucocytes). GI complaints were evaluated by a questionnaire. Results The intake of EcN resulted in a significantly lower increase in I-FABP and TBARS after exercise (p < 0.05). In contrast, no effect of EcN supplementation was found for hsCRP and leucocyte numbers. Similarly, no differences were found for levels of zonulin and claudin-3. Exercise-associated GI complaints were not affected by the probiotic supplement. Conclusion The probiotic EcN reduced the exercise-associated increase in oxidative stress. This antioxidative mechanism probably leads to a reduction of GI epithelial damage after exhaustive exercise. The lack of EcN effects on other markers of GI permeability and systemic inflammation is most likely due to an inadequate exercise load, with rather small and insignificant exercise effects on these parameters.

The present study investigated the hypothesis that dietary concentrations of leucine (Leu) in excess of the breeder´s recommendations activates protein synthesis and decreases protein degradation in muscle of broilers. Day-old male Ross 308 broilers (n = 450) were phase-fed corn-soybean meal-based diets during starter (d 1-10), grower (d 11-22), and finisher (d 23-34) period. The basal diets fed to the control group (L0) met the broilers' requirements for nutrients and amino acids, and contained Leu, Leu:isoleucine (Ile) and Leu:valine (Val) ratios, close to those recommended by the breeder (Leu:Ile: 100:54, 100:52, 100:51; Leu:Val 100:64, 100:61, 100:58; in starter, grower and finisher diet, resp.). Basal diets were supplemented with Leu to exceed the breeder's recommendations by 35% (group L35) and 60% (group L60). Growth performance during 34 d, and carcass weights, and breast and thigh muscle weights on d 34 were similar among groups. Hepatic and muscle mRNA levels of genes involved in the somatotropic axis [growth hormone receptor, insulin-like growth factor (IGF)-1, IGF binding protein 2, IGF receptor] on d 34 were not influenced by Leu. In the breast muscle, relative mRNA abundances of genes involved in the mammalian target of rapamycin (mTOR) pathway of protein synthesis (mTOR, ribosomal p70 S6 kinase) and the ubiquitin-proteasome pathway of protein degradation (F-box only protein 32, Forkhead box protein O1, Muscle RING-finger protein-1) on d 34 were largely similar among groups. Likewise, relative phosphorylation and thus activation of mTOR and ribosomal protein S6 involved in the mTOR pathway, and of eukaryotic translation initiation factor 2A (eIF2a) involved in the general control nonderepressible 2 (GCN2)/eIF2a pathway of protein synthesis inhibition, were not influenced. These data indicate that dietary Leu concentrations exceeding the broiler´s requirements up to 60% neither influence protein synthesis nor degradation pathways nor muscle growth in growing broilers.

SCOPE Sustainable protein sources are needed to meet the increasing protein demands of a continuously growing world population. This study is focused on the biotechnological production of a protein rich oyster mushroom (Pleurotus sajor-caju; PSC) by valorization of an agricultural side stream and the evaluation of the physiological effects of PSC in a rat model of metabolic syndrome. METHODS AND RESULTS PSC is produced via submerged cultivation in a 150 L bioreactor that utilizes isomaltulose molasses as its sole carbon source, and is further analyzed for its nutritional composition. A feeding trial is performed using Zucker rats which are fed a 5% PSC supplemented diet, for 4 weeks. Biochemical analyses reveal a significant reduction of the liver lipid concentrations and liver inflammation in the PSC fed obese rats in comparison to the obese rats from the control group. Hepatic qPCR analyses, differential transcript profiling, and enzyme activity measurements reveal a number of altered pathways that may be responsible for these anti-steatotic and anti-inflammatory effects of the mushroom. CONCLUSION Bioconversion of a low quality agricultural side stream to an improved protein source is performed by submerged cultured PSC, and the obtained mycelium shows strong anti-steatotic and anti-inflammatory effects.

This study investigated the hypothesis that dietary fats rich in lauric (C12) and myristic acid (C14) increase broiler performance and that the underlying mechanism involves antimicrobial effects on gut bacteria and changes in gut morphology. One hundred eighty 1-day-old Cobb-500 broilers were allotted to 3 groups. All groups received a basal diet consisting of maize, wheat, soybean meal, and a fat source (4.5, 7.0, 7.6, and 8.0% of fat product in the diet during d 1 to 9, 10 to 17, 18 to 27, and 28 to 35, respectively) until 35 d of age. The diet of the control group contained a fat with 67% of oleic and linoleic acid and 1.4% of C12 and C14 of total fatty acids, that of the esterified lauric and myristic acid (ELA) group a fat with 33% of esterified C12 and C14 and that of the free lauric and myristic acid (FLA) group a fat with 31% of both esterified and free (1:1) C12 and C14 (6 replicates/treatment, 10 birds/replicate). Gain and feed consumption did not differ between groups, but feed:gain was lower in FLA group as compared to the control group (P < 0.05). Carcass weight, liver weight, triglyceride content of liver and muscle, and muscle cholesterol were similar between groups; however, breast muscle weight was higher in the FLA than in the control group (P < 0.05). The villus height:crypt depth ratio of the duodenal wall did not differ between groups, but in the jejunum, it was lower in the FLA group as compared to the control group (P < 0.05). DNA copy numbers of Lactobacillus, Bifidobacteria, Enterobacteria, Escherichia coli, and Campylobacter jejuni in jejunal digesta were similar among groups. The study shows that dietary fats rich in free C12 and C14 improved feed:gain and breast muscle yield, but the observed effects could not be conclusively explained based on the parameters measured. The decreased jejunal villi:crypt ratio may point to changes in gut protein or cell turnover.

This study investigated the hypothesis that dietary supplementation of lignocellulose in broilers influences the gut bacterial population and bacterial fermentation, has anti-inflammatory effects, and increases mucin synthesis in the intestine, and, through these changes, influences broiler performance positively. Day-old male Cobb 500 broilers (n = 96) were allotted to 3 experimental groups and fed 3 different maize-wheat-soybean meal-based basal diets during days 1 to 10, 11 to 21, and 22 to 35. The basal diets were fed to the control group, and were supplemented with 0.8% of a standard lignocellulose (LCS) or a fermentable lignocellulose (LCF). Body weight and feed consumption were determined, and at slaughter (day 35), carcass and gizzard weights and gizzard content pH were recorded, and samples of jejunum, cecum, and colon mucosa and of cecum digesta were collected from 15 birds/group. Growth performance and feed intake were not influenced, but dressing percentage was higher in group LCF compared to the other groups. In group LCS and the control group, performance, gizzard weight and gizzard content pH, intestinal gene expression of pro-inflammatory cytokines and of the mucins 2, 5ac and 13, the cecal short-chain fatty acid (SCFA) profile, and bacterial diversity were similar, and relative abundance of bacterial groups (16S DNA sequencing) differed. Supplementation of LCF decreased the expression of the pro-inflammatory genes encoding interleukins 1s and 17 (P < 0.05) and those of 2 and 8 (P < 0.10) in the jejunum only. The bacterial population differed, and the SCFA profile shifted toward acetate at the expense of butyrate in group LCF compared to the control group. For example, the abundance of Firmicutes and of Ruminococcaceae and Lactobacillaceae decreased, whereas those of Peptostreptococcaceae, Erysipelotrichaceae, and Enterobacteriaceae and that of members of the phylum Proteobacteria increased in group LCF compared to the control group. These data indicate that the susceptibility of lignocellulose to fermentation is crucial for mediating its effects on intestinal gene expression and the bacterial population in the cecum, which may also affect dressing percentage.

The enhanced growth performance of calves fed a higher plane of nutrition pre-weaning is well documented, and the effect of butyrate on the development of the gastrointestinal tract in calves has been evaluated. The aim of this study was to examine the synergistic effects of ad libitum milk replacer (MR) feeding and butyrate supplementation on growth performance and energy metabolism in calves. Sixty-four (32 male, 32 female) Holstein calves were examined from birth until wk 11 of life. Calves received MR either ad libitum (Adl) or restrictively (Res) with (AdlB+, ResB+) or without (AdlB−, ResB−) 0.24% butyrate supplementation. Colostrum and transition milk were fed in predefined amounts (Res or Adl) for the first 3 d postpartum. Ad libitum and restrictive MR feeding with or without butyrate was performed from d 4 until wk 8 of age. From wk 9 to 10, all calves were gradually weaned and were fed 2 L/d until the end of the trial. Concentrate (CON), hay, and water were freely available. Intakes of MR and CON were measured daily. Calves were weighed at birth and weekly thereafter. Blood was drawn on d 1 before the first colostrum intake; on d 2, 4, and 7; and weekly thereafter until the end of the study to measure plasma concentrations of metabolites and hormones. Liver samples were taken at d 50 and at the end of the study to determine gene expression related to glucose metabolism. Milk, MR, and total nutrient intake were greater, but CON intake was lower in Adl than in Res calves, resulting in a greater body weight, but partially lower gain to feed ratio in Adl than in Res. Plasma concentrations of glucose and insulin were higher during the ad libitum milk-feeding period, whereas plasma β-hydroxybutyrate was lower in Adl than in Res. Plasma concentrations of nonesterified fatty acids, lactate, total bilirubin, and cortisol were lower, but triglyceride and cholesterol concentrations were higher in Adl than in Res at specific time points. Feed intake, growth performance, and metabolic and endocrine changes were insignificantly affected by butyrate, and hepatic gene expression of enzymes related to endogenous glucose production was barely influenced by ad libitum MR feeding and butyrate supplementation. Intensive MR feeding indicated greater stimulation of growth and anabolic metabolism, but butyrate supplementation did not further improve postnatal growth or anabolic processes either in intensive or restrictive MR-fed calves.

Scope The hypothesis is tested that insect meal, which has a low methionine content, reduces the hepatic phosphatidylcholine (PC):phosphatidylethanolamine (PE) ratio, which is a critical determinant of hepatic lipid synthesis, by decreasing availability of the methionine metabolite S-adenosylmethionine (SAM). Methods and results Obese rats (n = 24) are randomly divided into two groups (Obese Casein and Obese Insect) of 12 rats each. In addition, lean rats (n = 12) are used as control group (LC). Groups LC and OC receive a control diet with casein as protein source, whereas in the OI group, casein is replaced isonitrogenously by insect meal, which is found to be less digestible (-12% units). Plasma and liver concentrations of lipids and hepatic expression of lipid synthesizing genes are reduced in the OI group compared to the OC group. Plasma and liver concentration of PC and the PC:PE ratio are decreased in the OI group compared to the OC group, while hepatic concentration of SAM and the hepatic SAM:S-adenosylhomocysteine (SAH) ratio is lower in the OI group than in the OC group. Conclusion The decrease of the hepatic PC:PE ratio is probably a key mechanism explaining the pronounced antisteatotic and hypolipidemic action of insect meal in obese rats.

It was recently shown that niacin prevents the obesity-induced type I to type II fiber switching in skeletal muscle of obese rats and favors the development of a more oxidative metabolic phenotype and thereby increases whole body utilization of fatty acids. Whether niacin also causes type II to type I fiber switching in skeletal muscle of healthy rats has not been investigated yet. Thus, the present study aimed to investigate whether niacin supplementation influences fiber distribution and metabolic phenotype of different skeletal muscles with a distinct type I-to-type II fiber ratio in healthy rats.Twenty-four male, 10-week-old Sprague-Dawley rats were randomly assigned into two groups of 12 rats each and fed either a control diet with 30 mg supplemented niacin/kg diet (control group) or a high-niacin diet with 780 mg supplemented niacin/kg diet (high-niacin group).After 27 days of treatment, the percentage number of type I fibers in rectus femoris, gastrocnemius, and tibialis anterior muscles was 5-10% greater in the niacin group than in the control group, but did not differ between groups in soleus and vastus intermedius muscles. Transcript levels of genes encoding transcription factors regulating fiber switching, fiber-specific myosin heavy chain isoforms, and proteins involved in fatty acid utilization, oxidative phosphorylation, and angiogenesis did not differ between groups.The results show that niacin has only negligible effects on fiber distribution and its regulation as well as the metabolic phenotype of skeletal muscle in healthy rats.

Objective To evaluate medical efficacy and safety of crestal, minimally invasive sinus floor augmentation (MISFA) using an innovative method based on high hydraulic pressure. Study design Twenty MISFA using the novel Jeder-System were performed in 18 patients at 2 study sites in Vienna, Austria. The Jeder-System consists of the Jeder-drill, the Jeder-pump, and a connecting tube-set. The pump generates high hydraulic pressure (1.5 bar) pushing back the sinus membrane from the drill at the first perforation. The pump also monitors the whole procedure by constantly measuring pressure and volume. Results Five percent membrane perforation rate (1/20) only detected in the postoperative computed tomography scan and without implication for implant placement. Height gain of 9.2 ± 1.7 mm achieved (from 4.6 ± 1.4 mm to 13.8 ± 2.3 mm). Average patient satisfaction was 9.82 on scale from 1 to 10 (10 = very satisfied). Mean duration of sick leave was 0.19 days. 18-month survival rate was 95% (1/20 implant lost). Conclusions Within the limits of a prospective open cohort study with 20 cases, our data demonstrate the safety and medical efficacy of the novel method.

In the present study, we tested the hypothesis that niacin increases the oxidative capacity of muscle by increasing the oxidative type I muscle fiber content. Twenty-four obese Zucker rats were assigned to 2 groups of 12 rats that were fed either a control diet (O group) or a diet supplemented with 750 mg/kg diet niacin (O+N group) for 4 wk. In addition, one group of lean rats (L group) was included in the experiment and fed the control diet for 4 wk. Plasma and liver concentrations of TG were markedly greater in obese groups than in the L group but markedly lower in the O+N group than in the O group (P < 0.05). Rats of the O+N group had a higher percentage of oxidative type I fibers and higher mRNA levels of genes encoding regulators of muscle fiber composition (Ppard, Ppargc1a, Ppargc1b), angiogenic factors (Vegfa, Vegfb), and genes involved in fatty acid utilization (Cpt1b, Slc25a20, Slc22a4, Slc22a5, Slc27a1) and oxidative phosphorylation (Cox4i1, Cox6a2) and a higher activity of the mitochondrial oxidative enzyme succinate dehydrogenase in muscle than rats of the O and L groups (P < 0.05). These niacin-induced changes in muscle metabolic phenotype are indicative of an increased capacity of muscle for oxidative utilization of fatty acids and are likely mediated by the upregulation of Ppard, Ppargc1a, and Ppargc1b, which are key regulators of muscle fiber composition, mitochondrial biogenesis, angiogenesis, and genes involved in fatty acid catabolism and oxidative phosphorylation. The increased utilization of fatty acids by muscle might contribute to the strong TG-lowering effect of niacin treatment.

The study aimed to investigate the effects of chronic moderate exercise on regulation of intracellular calcium signaling as an important link to proliferation capacity in murine splenic T lymphocytes.Male CD1 Swiss mice were randomly assigned either to a control group (CG) or an exercise training group (EG). EG mice performed voluntary exercise for 3 months. Lymphocytes were isolated from murine spleens and intracellular calcium was determined by using Fura-2(AM) and fluorescence spectrometry. The combination of flow cytometry and carboxy-fluorescein succinimidyl ester labeling technique was used for determination of cell proliferation. The expression levels of Ca-regulating genes were determined by quantitative polymerase chain reaction (qPCR) analysis.Basal [Ca]i was significantly higher in mice from the EG compared with mice of the CG (P0.001, n = 6). Similarly, [Ca]i transients after stimulation with phytohemagglutinin, concanavalin A, and the anti-CD3 antibody induced were significantly increased in mice from the EG (P0.05, n = 5). However, no differences were found after stimulation with thapsigargin (P0.05, n = 5). CD3 T cells from EG showed higher mitogen-induced proliferation levels than from CG (P0.05/0.01, n = 5). The mRNA expression of cellular Ca-regulating genes, such as STIM1, Cav2.3, TRPV4, IP3R2, ORAI1, MCU, TRPM5, and TRPC1, were significantly downregulated (P0.05/0.01, n = 5).This study suggests that chronic moderate exercise improves intracellular Ca signaling in murine splenic lymphocytes. The enhanced availability of the second messenger Ca is followed by an improved cellular function such as cell proliferation. The downregulation of Ca homeostasis-related factor expression might be considered as a self-protective mechanism against elevated intracellular Ca signals.

High-intensity interval training (HIT) exercise has gained much interest in both performance and recreational sports. This study aims to compare the effect of HIT versus continuous (CONT) exercise with regard to changes of circulating T cells and progenitor cells.Subjects (n = 23) completed an HIT test and an isocaloric CONT test. Blood samples were collected before, immediately after, and 3 and 24 h postexercise for the assessment of low differentiated (CD3CD28CD57), highly differentiated T cells (CD3CD28CD57), regulatory T cells (Tregs) (CD4CD25CD127), hematopoietic progenitor cells (CD45CD34), and endothelial progenitor cells (CD45CD34KDR) by flow cytometry. The detection of apoptosis was performed by using labeling with annexin V. To analyze potential mechanisms affecting T cells, several hormones and metabolites were analyzed.Both exercise tests induced an increase of catecholamines, cortisol, and thiobarbituric acid-reactive substances (P0.05). CONT induced a higher increase of apoptosis in low differentiated T cells compared with the HIT (CONT: 3.66% ± 0.21% to 6.48% ± 0.29%, P0.05; HIT: 3.43% ± 0.31% to 4.71% ± 0.33%), whereas HIT was followed by a higher rate of apoptotic highly differentiated T cells (CONT: 21.45% ± 1.23% to 25.32% ± 1.67%; HIT: 22.45% ± 1.37% to 27.12% ± 1.76%, P0.05). Regarding Tregs, HIT induced a mobilization, whereas CONT induced apoptosis in these cells (P0.05). The mobilization of progenitor cells did not differ between the exercise protocols.These results suggest that HIT deletes mainly highly differentiated T cells known to affect immunity to control latent infections. By contrast, CONT deletes mainly low differentiated T cells and Tregs, which might affect defense against new infectious agents.

Supplementation of carnitine has been shown to improve performance characteristics such as protein accretion in growing pigs. The molecular mechanisms underlying this phenomenon are largely unknown. Based on recent results from DNA microchip analysis, we hypothesized that carnitine supplementation leads to a downregulation of genes of the ubiquitin proteasome system (UPS). The UPS is the most important system for protein breakdown in tissues, which in turn could be an explanation for increased protein accretion. To test this hypothesis, we fed sixteen male, four-week-old piglets either a control diet or the same diet supplemented with carnitine and determined the expression of several genes involved in the UPS in the liver and skeletal muscle. To further determine whether the effects of carnitine on the expression of genes of the UPS are mediated directly or indirectly, we also investigated the effect of carnitine on the expression of genes of the UPS in cultured C2C12 myotubes and HepG2 liver cells. In the liver of piglets fed the carnitine-supplemented diet, the relative mRNA levels of atrogin-1, E214k and Psma1 were lower than in those of the control piglets (P < 0.05). In skeletal muscle, the relative mRNA levels of atrogin-1, MuRF1, E214k, Psma1 and ubiquitin were lower in piglets fed the carnitine-supplemented diet than that in control piglets (P < 0.05). Incubating C2C12 myotubes and HepG2 liver cells with increasing concentrations of carnitine had no effect on basal and/or hydrocortisone-stimulated mRNA levels of genes of the UPS. In conclusion, this study shows that dietary carnitine decreases the transcript levels of several genes involved in the UPS in skeletal muscle and liver of piglets, whereas carnitine has no effect on the transcript levels of these genes in cultivated HepG2 liver cells and C2C12 myotubes. These data suggest that the inhibitory effect of carnitine on the expression of genes of the UPS is mediated indirectly, probably via modulating the release of inhibitors of the UPS such as IGF-1. The inhibitory effect of carnitine on the expression of genes of the UPS might explain, at least partially, the increased protein accretion in piglets supplemented with carnitine.

Scope: Metabolic stress induced by chronic high-fat (HF) diet feeding or genetically induced diabetes impairs carnitine status. Herein, we tested the hypothesis that regular endurance exercise (EE) improves the HF diet-induced impairment of carnitine status through stimulating the expression of hepatic genes involved in carnitine synthesis and uptake. Methods and results: Eighteen male C57BL/6 mice were assigned to three groups: group S received a standard diet, group HF received a HF diet, and group HF1EE received an HF diet and was regularly exercised on a treadmill. After 10wk, mice of the HF and the HF1EE groups were highly obese and insulin resistant compared with mice of the S group (po0.05), but mice of the HF1EE group were less insulin resistant than those of the HF group (po0.05). The HF group had lower carnitine concentrations and mRNA and protein levels of genes involved in carnitine synthesis and uptake in the liver than the S group (po0.05), whereas these parameters did not differ between the S group and the HF1EE group. Conclusion: These findings indicate that regular EE reverses an HF diet-induced impairment of hepatic carnitine content by stimulating hepatic carnitine synthesis and uptake.

Scope: Carnitine improves protein accretion, muscle mass, and protein:fat accretion in piglets. The underlying mechanisms, however, are largely unknown. Methods and results: To gain insight into mechanisms through which carnitine exerts these effects, we fed piglets either a control or a carnitine-supplemented diet, and analyzed the transcriptome in skeletal muscle. Carnitine concentrations in plasma and muscle were about four-fold higher in the carnitine group when compared to the control group. Transcript profiling revealed 211 genes to be differentially expressed in muscle by carnitine supplementation. The identified genes were mainly involved in molecular processes such as cytoskeletal protein binding, insulin-like growth factor (IGF) binding, transcription factor activity, and insulin receptor binding. Identified genes with the molecular function transcription factor activity encoded primarily transcription factors, most of which were down-regulated by carnitine, including pro-apoptotic transcription factors such as proto-oncogene c-fos, proto-oncogene c-jun and activating transcription factor 3. Furthermore, atrophy-related genes such as atrogin-1, MuRF1, and DRE1 were significantly down-regulated by carnitine. IGF signalling and insulin signalling were identified as significantly up-regulated regulatory pathways in the carnitine group. Conclusion: Carnitine may have beneficial effects on skeletal muscle mass through stimulating the anabolic IGF-1 pathway and suppressing pro-apoptotic and atrophy-related genes, which are involved in apoptosis of muscle fibers and proteolysis of muscle proteins, respectively.

In rodents, fasting increases the carnitine concentration in the liver by an up-regulation of enzymes of hepatic carnitine synthesis and novel organic cation transporter (OCTN) 2, mediated by activation of peroxisome proliferator-activated receptor (PPAR) alpha. This study was performed to investigate whether such effects occur also in pigs which like humans, as nonproliferating species, have a lower expression of PPARalpha and are less responsive to treatment with PPARalpha agonists than rodents. An experiment with 20 pigs was performed, which were either fed a diet ad-libitum or fasted for 24 h. Fasted pigs had higher relative mRNA concentrations of the PPARalpha target genes carnitine palmitoyltransferase 1 and acyl-CoA oxidase in liver, heart, kidney, and small intestinal mucosa than control pigs, indicative of PPARalpha activation in these tissues (P

Hepatic PPARalpha acts as the primary mediator of the adaptive response to fasting by upregulation of a number of genes involved in fatty acid catabolism. Whether carnitine-acylcarnitine translocase (CACT), which mediates the import of acylcarnitines into the mitochondrial matrix for subsequent beta-oxidation of fatty acid moieties, is also regulated by PPARalpha in the liver has not yet been investigated.Herein, we observed that hepatic mRNA abundance of CACT was increased by both, fasting and treatment with PPARalpha agonist WY-14,643 in wild-type mice but not PPARalpha-knockout mice (P0.05). Cell culture experiments revealed that CACT mRNA abundance was higher in liver cells treated with either WY-14,643 or PPARdelta agonist GW0742, but not with PPARgamma agonist troglitazone (TGZ) than in control cells (P0.05). In addition, reporter assays revealed activation of mouse CACT promoter by WY-14,643 and GW0742, but not TGZ. Moreover, deletion and mutation analyses of CACT promoter and 5'-UTR revealed one functional PPRE in the 5'-UTR of mouse CACT.CACT is upregulated by PPARalpha and PPARdelta, probably by binding to a functional PPRE at position +45 to +57 relative to the transcription start site. The upregulation of CACT by PPARalpha and PPARdelta, which are both important for the regulation of fatty acid oxidation in tissues during fasting, may increase the import of acylcarnitine into the mitochondrial matrix during fasting.

This study was performed to assess the effects of potato protein and fish protein on concentrations of lipids in plasma and lipoproteins and the expression of genes involved in lipid metabolism in pigs used as an animal model. Therefore, 27 young male pigs with an average body weight of 22 kg were fed diets supplemented with protein extracted from potatoes (containing 849 g protein/kg dry matter), Alaska Pollack fillet as a source of fish protein (containing 926 g crude protein/kg dry matter) or casein which was used as control, for 3 weeks. Diets were formulated to supply identical amounts of each protein to the pigs by the three protein sources, namely 116 g/day in first week and 150 g/day in the second and third week. Pigs fed potato protein had lower concentrations of cholesterol in plasma and LDL than pigs fed casein (p < 0.05); no effect was observed on concentrations of HDL cholesterol and triglycerides. Pigs fed fish protein had lower cholesterol concentrations in plasma, LDL and HDL, and lower triglyceride concentrations in triglyceride-rich lipoproteins than pigs fed casein (p < 0.05). mRNA concentrations of genes involved in bile acid synthesis and cholesterol uptake were higher in pigs fed fish protein than in pigs fed casein (p < 0.05); no effect on these genes was observed in pigs fed potato protein. Expression of genes involved in lipogenesis and fatty acid oxidation was not altered by fish protein. In conclusion, this study shows that fish protein and potato protein lower plasma cholesterol concentrations in pigs. The hypocholesterolaemic effect of fish protein might be in part caused by a stimulation of bile acid synthesis; the reason for the hypocholesterolaemic effect of potato protein requires further elucidation.

In contrast to other species, less is known about carnitine homeostasis in the pig. This study was performed to yield information about the site of carnitine synthesis and carnitine concentrations in various tissues of pigs (Sus scrofa). We found that among several pig tissues, a considerable activity of gamma-butyrobetaine dioxygenase (BBD), the last enzyme of carnitine synthesis, exists, like in humans and several other species, only in liver and kidney. Activity of that enzyme in liver and kidney was lower at birth than in the subsequent weeks of life. Highest carnitine concentrations were found in skeletal muscle and heart. Carnitine concentrations in plasma, liver and kidney at birth were higher than in the subsequent weeks of life in spite of the low BBD activity at birth. In conclusion, this study shows that liver and kidney are the major sites of carnitine synthesis and that neonatal pigs do not have an insufficient carnitine status.

For the investigation of the metabolism and biosynthesis of carnitine, sensitive determination of carnitine and its metabolic precursors, trimethyllysine and gamma-butyrobetaine, is required. We present here a new simplified method for the analysis of carnitine, its acetyl- and propyl esters, as well as trimethyllysine and gamma-butyrobetaine without need for derivatization reactions by means of normal-phase LC and electrospray ionization tandem mass spectrometry. The limits of quantification were between 5 nM for acetyl carnitine and 70 nM for carnitine. Relative standard deviations in a fivefold determination of standard solutions were between

Protein from white lupin is capable of lowering plasma lipids. We investigated in this study the effect of total protein extracts (TPEs) from different cultivars of blue lupin (Probor, Vitabor and Boregine) and alpha-/beta-conglutin from Boregine on the plasma lipids of rats. Rats were fed on a hypercholesterolemic diet containing either lupin protein (50 g/kg) or casein (50 g/kg) for 17 d. The rats fed with TPE from Vitabor and alpha-/beta-conglutin had lower triglyceride concentrations in the plasma (-24% and -21%, respectively) and very-low-density lipoprotein (VLDL; -40% and -29%, respectively) than the rats fed with casein. TPE from Vitabor was also capable of lowering low-density lipoprotein (LDL) cholesterol (-37%). In the liver of the rats fed with TPE from Vitabor, the expression of the genes involved in triglyceride and cholesterol synthesis was down-regulated. This study shows that the Vitabor cultivar of blue lupin had the most beneficial effect on plasma lipids which was presumed to have been caused by the down-regulation of genes involved in lipid synthesis.

Protein tyrosine phosphatase 1B (PTP1B) is a key enzyme in the counter-regulation of insulin signaling and in the stimulation of fatty acid synthesis. Selenium (Se), via the activities of glutathione peroxidase (GPx) and thioredoxin reductase (TrxR), is involved in the removal of H2O2 and organic peroxides, which are critical compounds in the modulation of PTP1B activity via glutathionylation. Our study with growing rats investigated how the manipulation of dietary Se concentration influences the regulation of PTP1B and lipogenic effects mediated by PTP1B. Weanling albino rats were divided into 3 groups of 10. The negative control group (NC) was fed a Se-deficient diet for 8 wk. Rats in groups Se75 and Se150 received diets supplemented with 75 or 150 mg Se/kg. Se supplementation of the rats strongly influenced expression and activity of the selenoenzymes cytosolic GPx, plasma GPx, phospholipidhydroperoxide GPx, and cytosolic TrxR, and liver PTP1B. Liver PTP1B activity was significantly higher in groups Se75 and Se150 than in the NC group and this was attributed to a lowered inhibition of the enzyme by glutathionylation. The increased liver PTP1B activity in groups Se75 and Se150 resulted in 1.1- and 1.4-fold higher liver triglyceride concentrations than in the NC rats. The upregulation of the sterol regulatory element binding protein-1c and of fatty acid synthase, 2 PTP1B targets, provided a possible explanation for the lipogenic effect of PTP1B due to the manipulation of dietary Se. We therefore conclude that redoxregulated proteins, such as PTP1B, represent important interfaces between dietary antioxidants such as Se and the regulation of metabolic processes. J. Nutr. 138: 2328‐2336, 2008.

It has been shown that some dietary plant proteins beneficially influence lipid metabolism in animals. The effect of pea protein in this respect however has not yet been investigated. Therefore, we studied the effect of purified pea protein on the lipid metabolism in rats. Twenty-four rats received diets with either 200 g/kg of casein or purified pea protein for 16 days. Concentrations of triacylglycerols in liver, plasma and lipoproteins did not differ between both groups of rats. However, rats fed the pea protein diet had a lower concentration of total cholesterol in the liver and the very low density lipoproteins (VLDL) fraction than rats fed the casein diet (p < 0.05); cholesterol concentration in plasma, low density lipoproteins (LDL) and high density lipoproteins (HDL) did not differ between both groups. Rats fed pea protein moreover had an increased mRNA concentration of cholesterol-7alpha-hydroxylase in the liver and an increased amount of bile acids excreted via faeces compared with rats fed casein (p < 0.05). Concomitantly, mRNA concentrations of sterol regulatory element-binding protein (SREBP)-2 and its target genes 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and LDL receptor in the liver were increased in rats fed pea protein (p < 0.05). The data of this study suggests that pea protein stimulates formation and excretion of bile acids, which leads to a reduced hepatic cholesterol concentration and a reduced secretion of cholesterol via VLDL. An increased gene expression of SREBP-2 and its target genes HMG-CoA reductase and LDL receptor may be a means to compensate for the increased loss of cholesterol for bile acid synthesis.

Activation of PPARalpha by clofibrate has recently been shown to cause upregulation of the high-affinity carnitine transporter novel organic cation transporter (OCTN) 2 in small intestine. This strongly suggests that PPARalpha activation in response to clofibrate treatment improves the absorption of carnitine from the diet. To test this hypothesis, we performed an experiment with rats which were fed diets with or without 5 g clofibrate/kg diet and with or without 5 g L-carnitine/kg diet. PPARalpha was significantly activated by clofibrate in small intestine as evidenced by increased relative mRNA concentrations of the PPARalpha target gene acyl-CoA oxidase (P0.05). Relative mRNA concentration of OCTN2 in small intestine was significantly increased by clofibrate (P0.05) but not the carnitine supplementation, whereas relative mRNA concentrations of other carnitine transporters (OCTN1, ATB(0+)) in small intestine were not influenced by either clofibrate or carnitine. The absorption rate of carnitine in small intestine was markedly higher in rats treated with clofibrate than in those treated without clofibrate (P0.05). In conclusion, the present study shows that administration of clofibrate to rats increases carnitine absorption in small intestine which is probably due to the observed upregulation of OCTN2 mediated by activation of PPARalpha.

Recent studies have shown that treatment of rodents with agonists of peroxisome proliferator-activated receptor (PPAR)-alpha causes an up-regulation of novel organic cation transporter (OCTN)-2, a carnitine transporter, and increases carnitine concentration in the liver. This study was performed to investigate whether such effects occur also in pigs which like humans have a lower expression of PPAR alpha and are less responsive to treatment with PPAR alpha agonists than rodents. An experiment with 18 pigs was performed which were fed a control diet or the same diet supplemented with 5 g clofibrate/kg for 28 days. Pigs treated with clofibrate had higher relative mRNA concentrations of OCTN2 in liver (3.1-fold), skeletal muscle (1.5-fold) and epithelial cells from small intestine (1.8-fold) than control pigs (P

It has been shown that treatment of rats with clofibrate, a synthetic agonist of PPARα, increases mRNA concentration of organic cation transporters (OCTN)-1 and -2 and concentration of carnitine in the liver. Since oxidised fats have been demonstrated in rats to activate hepatic PPARα, we tested the hypothesis that they also up regulate OCTN. Eighteen rats were orally administered either sunflower-seed oil (control group) or an oxidised fat prepared by heating sunflower-seed oil, for 6 d. Rats administered the oxidised fat had higher mRNA concentrations of typical PPARα target genes such as acyl-CoA oxidase, cytochrome P450 4A1 and carnitine palmitoyltransferases-1A and -2 in liver and small intestine than control rats (P P P

Activation of PPARalpha by clofibrate has recently been shown to cause upregulation of carnitine transporter organic cation transporter (OCTN) 2 and elevated carnitine concentrations in rat liver. The present study has been conducted to further explore the effect of clofibrate on OCTN expression, carnitine biosynthesis, and carnitine accumulation in different rat tissues, and thus two groups of rats were fed diets containing 0.5% clofibrate or 0% clofibrate (control group). PPARalpha-responsive genes were markedly upregulated in the liver (P

Trans-fatty acid intake is associated with an increased risk of CHD and diabetes. The effects of singletrans-fatty acid isomers are largely unexplored. The present study examined the effects of a 6-week supplementation with twotrans-18 : 1 isomers (trans-11 andtrans-12) in human subjects on immune cells, several inflammatory and immunological biomarkers (for example, IL, TNFα, C-reactive protein, adiponectin, intercellular adhesion molecule-1, prostacyclin, phagocytic process). Following a 2-week adaptation period without supplements, the test group (n12) received vaccenic acid (trans-11-18:1) andtrans-12-18 : 1 in equal amounts (6·0 g/d) for 6 weeks. The control group (n12) consumed an oil withouttrans-fatty acids and conjugated linoleic acids (CLA). Samples were collected at the end of both periods.Trans-11- andtrans-12-18 : 1 were significantly increased in cellular lipids. The endogenous synthesis ofcis-9,trans-11-CLA fromtrans-11-18 : 1 was demonstratedviaincreased CLA in cellular lipids of the test group. Generally,trans-isomer supplementation did not affect either inflammatory biomarkers (for example, IL-6, IL-8, TNFα) or immune function (for example, phagocytosis) during the present study. The dietary supplementation oftrans-11- andtrans-12-18 : 1 (6 g/d) and their accumulation in leucocytes had no effects on biomarkers of inflammation and immune function. However, because of the limited data on the safety oftrans-fatty acid intake and effects of individualtransisomers on human health (for example,trans-9-18 : 1,trans-10-18 : 1) at present, it is prudent to reducetrans-fat intake in general.

Recent studies have shown that dietary oxidised fats influence the lipid metabolism in rats by activation of PPARα. In this study, we investigated whether a mildly oxidised fat causes activation of PPARα in pigs which are non-proliferators like man. Eighteen pigs were assigned to two groups and received either a diet containing 90 g/kg of a fresh fat or the same diet with 90 g/kg of an oxidised fat prepared by heating for 24 h at 180°C in a deep fryer. Pigs fed the oxidised fat had a higher peroxisome count, a higher activity of catalase and a higher mRNA concentration of mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase in the liver and a higher concentration of 3-hydroxybutyrate in plasma than pigs fed the fresh fat (P P

Soy intake acts hypolipidemically. Besides isoflavones, soy protein itself is suggested to influence plasma lipid concentrations. We investigated the effects of an alcohol-washed isoflavone-poor soy protein isolate on plasma and liver lipids and the hepatic expression of genes encoding proteins involved in cholesterol and fatty acid metabolism. Therefore, rats were fed diets containing 200 g/kg of either ethanol-extracted soy protein isolate or casein over 22 days. Rats fed soy protein isolate had markedly lower concentrations of liver cholesterol and lower concentrations of triglycerides in the liver and in plasma than rats fed casein (P.05). Rats fed soy protein isolate had lower relative mRNA concentrations of sterol-regulatory element-binding protein (SREBP)-2, 3-hydroxy-3-methylglutaryl coenzyme A reductase, low-density lipoprotein receptor, cholesterol 7alpha-hydroxylase, apolipoprotein B, Delta9-desaturase and glucose-6-phosphate dehydrogenase in the liver than rats fed casein (P.05). Hepatic mRNA concentration of SREBP-1c tended to be lower in rats fed soy protein isolate (P.10). Hepatic mRNA concentrations of insulin-induced gene (Insig) 1 and Insig-2 and of microsomal triglyceride transfer protein, as well as plasma concentrations of free fatty acids, insulin and glucagon, were not different between the two groups. In conclusion, this study suggests that isoflavone-poor soy protein isolate affects cellular lipid homeostasis by the down-regulation of SREBPs and its target genes in the liver, which are involved in the synthesis of cholesterol and triglycerides.

To elucidate the mechanisms underlying the cholesterol lowering effects of PPARalpha agonists we investigated key regulators of cholesterol synthesis and uptake in rats and in the rat hepatoma cell line Fao after treatment with the PPARalpha agonists clofibrate and WY 14,643, respectively. In rat liver as well as in Fao cells, PPARalpha activation led to a decrease of transcriptionally active nuclear SREBP-2. mRNA concentrations of the key regulators of SREBP processing, Insig-1 in rat liver and Insig-1 and Insig-2a in Fao cells, were increased upon PPARalpha activation. Thus we suggest, that the observed reduction of the amount of nuclear SREBP-2 was due to an inhibition of the processing of the precursor protein. Both, in rat liver and in Fao cells, mRNA concentrations of the SREBP-2 target genes HMG-CoA reductase (EC1.1.1.34) and LDL receptor were reduced after treatment with the PPARalpha agonists. Furthermore, treatment of Fao cells with WY 14,643 reduced cholesterol synthesis. As a result, the amount of total cholesterol in liver, plasma and lipoproteins of clofibrate treated rats and in WY 14,643 treated Fao cells was decreased compared to control animals and cells, respectively. In conclusion, we could show a novel link between PPARalpha and cholesterol metabolism by demonstrating that PPARalpha activation lowers cholesterol concentration by reducing the abundance of nuclear SREBP-2.

Background: Recently, it has been shown that dietary lupin protein lowers plasma triglyceride concentrations in rats. In this study, we investigated the hypothesis that this effect is due to a downregulation of sterol regulatory element-binding protein (SREBP)-1c, a transcription factor that regulates the expression of lipogenic enzymes in the livers of rats. Methods: Two groups of 12 rats each were fed semisynthetic diets containing 200 g/kg of either casein (control group) or lupin protein from Lupinus albus for 22 days. Results: Rats fed the diet containing lupin protein had lower concentrations of triglycerides in the liver, plasma and VLDL + chylomicrons (p < 0.05). The concentration of protein in VLDL + chylomicrons was also lower in rats fed lupin protein than in rats fed casein (p < 0.05). The mRNA concentrations of SREBP-1c and fatty acid synthase in the liver were lower in rats fed lupin protein than in rats fed casein (p < 0.05). The mRNA concentrations of lipoprotein lipase in the liver did not differ between both groups of rats. Conclusion: This study confirms that a protein isolated from L. albus is strongly hypotriglyceridemic in rats. It is shown for the first time that this effect is at least in part due to a downregulation of SREBP-1c in the liver which in turn leads to a reduction in hepatic fatty acid synthesis.

The present study was performed to assess the bioefficacy of DL-methionine hydroxy analogue-free acid (MHA) in comparison to DL-methionine (DLM) as sources of methionine for growing male white Pekin ducks in the first 3 wk of life. For this aim, 580 1-day-old male ducks were allocated into 12 treatment groups and received a basal diet that contained 0.29% of methionine, 0.34% of cysteine and 0.63% of total sulphur containing amino acids or the same diet supplemented with either DLM or MHA in amounts to supply 0.05, 0.10, 0.15, 0.20, and 0.25% of methionine equivalents. Ducks fed the control diet without methionine supplement had the lowest final body weights, daily body weight gains and feed intake among all groups. Supplementation of methionine improved final body weights and daily body weight gains in a dose dependent-manner. There was, however, no significant effect of the source of methionine on all of the performance responses. Evaluation of the data of daily body weight gains with an exponential model of regression revealed a nearly identical efficacy (slope of the curves) of both compounds for growth (DLM = 100%, MHA = 101%). According to the exponential model of regression, 95% of the maximum values of daily body weight gain were reached at methionine supplementary levels of 0.080% and 0.079% for DLM and MHA, respectively. Overall, the present study indicates that MHA and DLM have a similar efficacy as sources of methionine for growing ducks. It is moreover shown that dietary methionine concentrations of 0.37% are required to reach 95% of the maximum of daily body weight gains in ducks during the first 3 wk of life.

Polyphenol-rich plant products as feed supplements have been shown to exert beneficial effects on feed efficiency in piglets. However, tannins as components of polyphenol-rich plant products are able to reduce the absorption of various trace elements. The present study investigated the effect of two polyphenol-rich dietary supplements, grape seed and grape marc meal extract (GME) and spent hops (SH), on iron (Fe), zinc (Zn) and copper (Cu) status in piglets supplied adequately with those trace elements. A trial with three groups of piglets which received a Control diet or the same diet supplemented with either 1% GME or 1% SH over a period of 4 weeks was performed. Concentrations of Fe, Zn and Cu in plasma, total iron binding capacity and saturation of transferrin in plasma did not differ between the three groups. Piglets fed the diet supplemented with SH showed no differences in the concentrations of Fe, Zn and Cu in the liver in comparison to the Control group. Piglets fed the diets supplemented with GME showed slightly lower concentrations of Zn and Cu in the liver than Control piglets (p < 0.05); however, concentrations of both elements remained in the physiological range. Overall, this study shows that the polyphenol-rich plant products GME and SH had marginal effect on the status of Fe, Zn and Cu in piglets.

The present study aimed to determine the nutrient and energy content of fresh and ensiled grape pomace (GP) from different grape varieties originating from Germany, and to estimate the feed value of dried white, dried red and ensiled white GP by calculating nutrient digestibility and the content of metabolisable energy (ME) and net energy lactation (NEL) measured in sheep as a ruminant model. GP from red cultivars had higher contents of organic matter (OM), crude protein (CP), ether extract (EE), crude fibre (CF), total phenolic contents (TPC) and ME, whereas the concentrations of ash and sugar were lower than from white cultivars. Compared with untreated GP, ensiled GP had increased concentrations of CP (+19%), ether extract (EE; +23%) and CF (+12%) and a higher ME content (+7%) and markedly decreased concentrations of sugar (-99.6%) and TPC (-48%). The concentrations of dry matter, OM and ash were not different between ensiled and fresh GP. Compared with dried GP, ensiled GP had a higher nutrient digestibility (OM, +32%; CP, +43%; CF, +46%; neutral detergent fibre [NDF], +54%; acid detergent fibre [ADF], +69%) and higher energy values (ME, +16%; NEL, +19%). The digestibility of OM, CP, EE and CF and the energy content were higher for dried red than for dried white GP, whereas the digestibility of NDFOM and ADFOM was lower for dried red than dried white GP. In conclusion, the results show that both red and white GP are suitable dietary sources for enrichment with TPC. Furthermore, compared with drying ensiling of GP improves the feeding value of GP and is a good possibility of preserving the seasonally produced by-product of winemaking for ruminant feeding.

Background and Purpose Since patients with phenylketonuria (PKU) have to follow a lifelong restriction of natural protein to lower phenylalanine-intake, they never eat fish. This diet may lead to a chronic deficit of omega-3 and omega-6 fatty acids with the risk of early atherosclerotic changes. The aim of the study was to analyse the fatty acid profile of PKU patients and to correlate the results with surrogate markers of early atherosclerotic changes [enhanced carotid intima media thickness (CIMT) and s-stiffness index] and platelet activation. Methods In 43 PKU patients and in 58 healthy controls we prospectively examined the fatty acid profile, CIMT, s-stiffness index and platelet activation (flow cytometric determination of markers of platelet activation). CIMT was measured bilaterally by ultrasound. CIMT mean was defined as the mean value of the sum of CIMT left and CIMT right. Results Despite of lower HDL-cholesterol and higher triglyceride concentrations in the PKU group, there was no significant difference in the omega-6 or omega-3 fatty acid profile, CIMT, s-stiffness index between both groups. Platelet activation was not enhanced in the PKU group. Conclusions Fish-free diet does not induce early atherosclerotic changes or enhanced platelet activation in PKU patients.

Morphological and metabolic adaptations of the human skeletal muscle to exercise are crucial to improve performance and prevent chronic diseases and metabolic disorders. In this study we investigated human skeletal muscle protein composition in endurance trained (ET) versus untrained individuals (UT) and its modulation by an acute bout of endurance exercise. Participants were recruited based on their VO 2 max and subjected to a bicycle exercise test. M. vastus lateralis biopsies were taken before and three hours after exercise. Muscle lysates were analyzed using off-gel LC-MS/MS. Relative protein abundances were compared between ET and UT at rest and after exercise. Comparing UT and ET, we identified 92 significantly changed proteins under resting conditions. Specifically, fiber-type-specific and proteins of the oxidative phosphorylation and tricarboxylic acid cycle were increased in ET. In response to acute exercise, 71 proteins in ET and 44 in UT were altered. Here, a decrease of proteins involved in energy metabolism accompanied with alterations of heat shock and proteasomal proteins could be observed. In summary, long-term endurance training increased the basal level of structural and mitochondrial proteins in skeletal muscle. In contrast, acute exercise resulted in a depletion of proteins related to substrate utilization, especially in trained athletes. Biological significance The investigation of the human skeletal muscle proteome in response to exercise may provide novel insights into the process of muscular plasticity. It is of importance in the development of exercise-based strategies in the prevention and therapy of many chronic inflammatory and degenerative diseases which are often accompanied by muscular deconditioning. Up to date, proteomic investigations of the human muscle proteome in adaptation to exercise are mainly focused on untrained individuals and often restricted to animal studies. In the present study we compare the protein composition in endurance trained athletes and untrained individuals in the resting muscle and its modulation in response to acute exercise. To our knowledge, we present the first comprehensive analysis of skeletal muscle proteome alterations in response to acute and long-term exercise intervention.

Objective We assessed the effect of dietary proteins isolated from beef, pork, and turkey meat on concentrations of cholesterol and triacylglycerols in plasma, lipoproteins, and liver and the composition of the microsomal membrane (fatty acids, phosphatidylcholine/phosphatidylethanolamine ratio) compared with that of casein and soy protein in rats. Methods Five groups of 12 rats each were fed semisynthetic diets for 20 d that contained 200 g/kg of proteins isolated from beef, pork, or turkey meat or, as controls, casein or soy protein. Results Rats fed beef, pork, or turkey proteins did not differ in cholesterol concentrations of plasma, lipoproteins, and liver and in composition of microsomal membrane from rats fed the casein diet. All groups fed a protein from an animal source had higher very low-density lipoprotein (VLDL) and liver cholesterol concentrations than did rats fed soy protein. However, rats fed pork protein had lower concentrations of triacylglycerols in liver, plasma, and VLDL and lower mRNA concentrations of sterol regulatory element binding protein-1 and glucose-6-phosphate dehydrogenase than did rats fed casein. However, concentrations of plasma and VLDL triacylglycerols in rats fed pork protein were not as low as those observed in rats fed soy protein. Conclusion Proteins isolated from beef, pork, or turkey meat do not differ from casein in their effects on cholesterol metabolism. Pork protein decreases plasma triacylglycerol concentrations compared with casein but not compared with soy protein. The triacylglycerol-lowering effect of pork protein compared with casein is suggested to be caused by decreased hepatic fatty acid synthesis.

It has been shown that clofibrate treatment increases the carnitine concentration in the liver of rats. However, the molecular mechanism is still unknown. In this study, we observed for the first time that treatment of rats with the peroxisome proliferator activated receptor (PPAR)-alpha agonist clofibrate increases hepatic mRNA concentrations of organic cation transporters (OCTNs)-1 and -2 which act as transporters of carnitine into the cell. In rat hepatoma (Fao) cells, treatment with WY-14,643 also increased the mRNA concentration of OCTN-2. mRNA concentrations of enzymes involved in carnitine biosynthesis were not altered by treatment with the PPARalpha agonists in livers of rats and in Fao cells. We conclude that PPARalpha agonists increase carnitine concentrations in livers of rats and cells by an increased uptake of carnitine into the cell but not by an increased carnitine biosynthesis.