Anti-CD20 antibody therapy and risk of infection in patients with demyelinating diseases.

BACKGROUND: Anti-CD20 antibody therapy may be associated with an increased risk of infections. We therefore investigated risk factors for infection in patients with demyelinating diseases treated with anti-CD20 antibody therapy. METHODS: In this retrospective uncontrolled study, patients ever treated with anti-CD20 antibodies at an academic clinic were identified through the Danish Multiple Sclerosis Registry (DMSR). Data were collected from medical charts and the DMSR. We assessed occurrence of severe infections (requiring hospitalization), varicella zoster virus (VZV), major comorbidities and routine laboratory values for lymphocytes, IgG and IgM. RESULTS: A total of 447 patients ever treated with anti-CD20 antibody therapy were identified; of these 416 with 649 patient years of follow-up were still under therapy. In this group, seven patients had VZV infections, and 16 patients had been hospitalized with infections during up to three years of follow-up on anti-CD20 therapy. Comorbidity was recorded in 80 patients. The risk of severe infection was associated with comorbidities, higher age, longer duration of treatment, and higher Expanded Disability Status Scale (EDSS) scores. In multivariable analyses treatment duration, EDSS scores and presence of comorbidity were independently associated with risk of severe infections. Serum concentrations of IgG and IgM decreased with increasing duration of therapy but were not associated with risk of severe infections. Patients with VZV infection had lower lymphocyte counts and lower serum concentrations of IgM. In multivariable analyses only lymphocyte counts were independently associated with risk of VZV infection. CONCLUSIONS: In this retrospective study of patients treated with anti-CD20 antibodies, the risk of infections requiring hospitalization was independently associated with comorbidities, duration of treatment, and higher EDSS scores. Risk of VZV infection was independently associated with lymphopenia. Future studies investigating strategies for mitigating risk of infection in patients treated with anti-CD20 antibodies are warranted, especially for older patients, patients with higher levels of disability and for patients with a longer duration of treatment.

Mammary nutrient uptake in multiparous sows fed supplementary arginine during gestation and lactation.

Arginine is the precursor for the synthesis of nitric oxide and may increase mammary plasma flow (MPF), which may in turn increase mammary nutrient uptake. Quantifying mammary nutrient uptake improves our understanding of mammary nutrient metabolism and may potentially allow identification of limiting nutrients for colostrum and milk production. Thus, the objectives of the present study were 1) to study the impact of 25 g/d of crystalline Arg (ARG) on MPF and uptake of nutrients by the mammary glands compared with an isonitrogenous supply of Ala (51 g/d; control [CON]) fed to a total of 8 sows from d 30 of gestation until weaning on d 28 of lactation and 2) to quantify mammary nutrient uptake in late gestation and in early and at peak lactation. Sows were surgically fitted with indwelling catheters on d 76 ± 2 SEM of gestation. -amino hippuric acid (AH) was infused (3.0 mmol/h) in the infusion catheter inserted in the mammary vein, initiated 1 h before the first blood sample at -10, -3, 3, and 17 d in milk (DIM). Blood samples were simultaneously drawn from catheters inserted in the femoral artery and the mammary vein, and the samples were collected in hourly intervals from 0.5 h before to 6.5 h after feeding. Sow milk production was assessed at 3 and 17 DIM. Arterial plasma concentrations of Arg and Ala were increased in ARG and CON sows, respectively ( < 0.01), whereas we did not succeed in detecting a greater MPF in ARG sows ( = 0.30). Arterial-venous differences ( = 0.03) and net mammary flux ( = 0.01) of Ala were increased in CON sows, while the net flux of most other metabolites ( > 0.05) was unaffected by treatment. The mammary extraction of all essential AA was below 13% in late gestation. The average mammary extraction of essential AA at peak lactation was greatest for Leu (51%), while the preprandial extraction was greatest for Lys (57%). The mammary carbon balance (input-output) was negative (-39 ± 12 mol C/d) in early lactation but almost balanced at peak lactation (-13 ± 14 mol C/d), suggesting that mammary fat depots contributed to milk synthesis. In conclusion, we failed to observe an increased MPF and mammary uptake of AA and energy metabolites in ARG-supplemented sows. The mammary extraction rate of essential AA indicated that AA were not limiting for the mammary glands in late gestation, while Lys and Leu appeared to be the 2 most limiting essential AA for milk production at peak lactation.

Supplementing newborn intrauterine growth restricted piglets with a bolus of porcine colostrum raises rectal temperatures one degree Celsius.

Hyperprolific sows have increased litter sizes but also result in more piglets that have been exposed to intrauterine growth restriction (IUGR). These IUGR piglets are likely to have a low rectal temperature and lower blood glucose levels compared with normal piglets at birth. Therefore, we hypothesized that a colostrum bolus at birth and/or heat from an external source would have a positive effect on blood glucose levels, rectal temperatures, and growth up to 8 h postpartum. In addition, liver glycogen and blood values at 8 h were investigated. Eighty-four piglets were classified at birth (time = 0) as IUGR based on their head morphology and randomly allocated to 1 of 4 treatments ( = 21) in a 2 × 2 factorial arrangement: 1) with or without a porcine colostrum bolus (12 mL/kg BW at birth) and 2) with sow or isolated from sow with external heat. Piglets were removed from the sow before they had suckled and were numbered and dried, and initial whole-blood glucose, rectal temperature, and BW were recorded. Piglets in the 2 treatments isolated from sow were placed under a heating lamp (150 W) with a temperature range of 35 to 39°C. Rectal temperatures, glucose, and BW were measured again at 1, 2, 4, 6, and 8 h after birth, and a final plasma sample and organs (liver and brain) were removed at 8 h. There was a time × colostrum bolus interaction ( = 0.026) and a time × sow interaction ( < 0.001) for whole-blood glucose. The piglets that were given a bolus had greater glucose levels after 1 h postpartum (time = 1 h) than piglets without a bolus at birth, but from time = 2 h and onward, there was no difference ( > 0.05). There was a time × colostrum bolus interaction ( < 0.001) and a time × sow interaction ( < 0.001) on rectal temperatures. One hour after birth, the piglets with a bolus had a greater rectal temperature compared with piglets without a bolus (37.5 vs. 36.6°C; < 0.001) and the piglets that had been isolated from the sow had a greater rectal temperature compared with the 2 treatments with sows (37.8 vs. 36.3°C; < 0.001). Four hours after birth, rectal temperature was not affected by treatments. In conclusion, both heat and a colostrum bolus increased rectal temperature by 1°C an hour after birth. However, after 4 h, no differences were found between the treatments. Interventions to help IUGR piglets postpartum most likely need to be frequent to have any effect on whole-blood glucose, rectal temperatures, and BW over the first 8 h.

Validation of biomarkers for loin meat quality (M. longissimus) of pigs.

The aim of this study was to validate previously reported associations between microarray gene expression levels and pork quality traits using real-time PCR. Meat samples and meat quality data from 100 pigs were collected from a different pig breed to the one tested by microarray (Large White versus Pietrain) and a different country of origin (Denmark versus Germany). Ten genes (CARP, MB, CSRP3, TNNC1, VAPB, TNNI1, HSPB1, TNNT1, TIMP-1, RAD-like) were chosen from the original microarray study on the basis of the association between gene expression levels and the meat quality traits meat %, back fat, pH24, drip loss %, colour a*, colour b*, colour L*, WB-SF, SFA, MUFA, PUFA. Real-time PCR detection methods were developed for validation of all ten genes, confirming association with drip loss (two of two genes), ultimate pH (three of four genes), a* (redness) (two of six genes) and L*(lightness) (two of four genes). Furthermore, several new correlations for MUFA and PUFA were established due to additional meat quality trait information on fatty acid composition not available for the microarray study. Regression studies showed that the maximum explanation of the phenotypic variance of the meat quality traits was 50% for the ultimate pH trait using these ten genes only. Additional studies showed that the gene expression of several of the genes was correlated with each other. We conclude that the genes initially selected from the microarray study were robust, explaining variances of the genes for the meat quality traits.

Caffeic acid, naringenin and quercetin enhance glucose-stimulated insulin secretion and glucose sensitivity in INS-1E cells.

AIMS: Caffeic acid, naringenin and quercetin are naturally occurring phenolic compounds (PCs) present in many plants as secondary metabolites. The aim of this study was to investigate their effect on glucose-stimulated insulin secretion (GSIS) in INS-1E cells and to explore their effect on expression of genes involved in ß-cell survival and function under normoglycaemic and glucotoxic conditions. METHODS: For acute studies, INS-1E cells were grown in 11 mM glucose (72 h) and then incubated with the PCs (1 h) with 3.3/16.7 mM glucose; whereas, for chronic studies, the cells were grown in 11 mM glucose (72 h) with/without the PCs, and then incubated with 3.3/16.7 mM glucose (1 h); thereafter, GSIS was measured. For GSIS and gene expression studies (GES) under glucotoxic conditions, two sets of cells were grown in 11/25 mM glucose with/without the PCs (72 h): one was used for GES, using real time RT-PCR, and the other was exposed to 3.3/16.7 mM glucose, followed by measurement of GSIS. RESULTS: The study demonstrated that the PCs can enhance GSIS under hyperglycaemic and glucotoxic conditions in INS-1E cells. Moreover, these compounds can differentially, yet distinctly change the expression profile of genes [Glut2 (glucose transporter 2), Gck (glucokinase), Ins1 (insulin 1), Ins2, Beta2 (neurogenic differentiation protein 1), Pdx1 (pancreatic and duodenal homeobox protein 1), Akt1 (RAC-α serine/threonine-protein kinase encoding gene), Akt2 (RAC-ß serine/threonine-protein kinase encoding gene), Irs1 (insulin receptor substrate 1), Acc1 (acetyl CoA carboxylase 1), Bcl2 (ß-cell lymphoma 2 protein), Bax (Bcl-2 associated X protein), Casp3 (Caspase 3), Hsp70 (heat shock protein 70), and Hsp90] involved in ß-cell stress, survival and function. CONCLUSION: The results indicate that the PCs tested enhance GSIS and glucose sensitivity in INS-1E cells. They also modulate gene expression profiles to improve ß-cell survival and function during glucotoxicity.

Intrauterine growth restricted piglets defined by their head shape ingest insufficient amounts of colostrum.

The increasing litter sizes of modern pig breeds have led to a significant number of piglets that are born undersized ("small" piglets) and some have been exposed to different degrees of intrauterine growth restriction (IUGR). The aim of this study was to investigate the physiology and capability to ingest colostrum of these small piglets, suffering from various degrees of IUGR, to see if their IUGR score could be a useful tool for easy identification of piglets in need of intervention in the colostrum period. Piglets were classified at birth based on head morphology. Piglets were classified either "normal," "mildly IUGR" (m-IUGR), or "severe IUGR" (s-IUGR), based on head morphology. Blood samples were collected at birth and at 24 h, and colostrum intake during two 12-h periods and blood metabolites at 0 and 24 h were measured. At 24 h, piglets weighing <900 g at birth and the median piglet in birth order were sacrificed, and organ weights and hepatic glycogen were measured. Overall, there was an influence of the piglets' classification on most characteristics, with normal piglets having a greater colostrum intake between 0 and 12 h (P < 0.001) and between 12 and 24 h (P < 0.05), and higher birth weight, crown rump length, body mass index, and ponderal index (P < 0.001), and a tendency toward a higher vitality score (P < 0.069) than s-IUGR piglets. There was a time × IUGR interaction, with plasma glucose levels being lowered (P < 0.001) and lactate levels elevated (P < 0.001) in s-IUGR piglets at 24 h compared with normal and m-IUGR piglets. Some differences were found in electrolytes; sodium plasma concentrations were greatest for normal piglets (P < 0.05) and highest at 0 h (P < 0.05). At 24 h of age, s-IUGR piglets had a higher heart (P < 0.001) and brain percentage (P < 0.001), and a lower liver percentage (P < 0.001) relative to body weight, compared with normal piglets. In addition, s-IUGR piglets had less hepatic glycogen than m-IUGR piglets and normal piglets. The present study showed that the physiology of piglets in the colostrum period was affected by IUGR status at birth and their intermediary metabolism was altered due to different colostrum intakes. Furthermore, it was demonstrated that the head shape of newborn piglets is a good selection criteria for identifying piglets that need oral supplementation during the neonatal stage.

Novel aspects of health promoting compounds in meat.

Meat is an integral part of the human diet. Besides essential amino acids and nutritive factors of high quality and availability, meat provides often overlooked components of importance for human health. These are amino acids and bioactive compounds that may be very important in i) preventing muscle wasting diseases, such as in sarcopenia, ii) reducing food and caloric intake to prevent metabolic syndrome, iii) blood pressure homeostasis via ACE-inhibitory components from connective tissue, and iv) maintaining functional gut environment through meat-derived nucleotides and nucleosides. In addition, meat could be an important source of phytanic acid, conjugated linoleic acids and antioxidants. Further, it becomes increasingly apparent that design of in vitro meat will be possible, and that this development may lead to improved health benefits from commercially viable and sustainable meat products.

Meat Science and Muscle Biology Symposium: in utero nutrition related to fetal development, postnatal performance, and meat quality of pork.

Intrauterine growth restriction (IUGR) occurs naturally in pigs and leads to low birth weight of piglets due to undernutrition caused by placental insufficiency. For 2 main reasons, low birth weight causes economic loss. First, low birth weight pigs have a greater mortality and increasing the litter size causes more low birth weight piglets within litters. Second, surviving low birth weight piglets have reduced performance (i.e., ADG, feed conversion rate, and percentage meat). To develop dietary strategies for preventing IUGR, knowledge of the biological basis of IUGR is required. Muscle fiber number, formed during myogenesis, is correlated positively with performance traits and has been shown in several studies to be reduced in low birth weight pigs. Postnatal muscle hypertrophy is due to satellite cell number per fiber at birth and their rate of proliferation as well as protein deposition (i.e., protein synthesis and degradation). Previous studies and some recent ones indicate that low birth weight littermates in mice are born with fewer satellite cells and studies on pigs show that the rate of satellite cell proliferation may vary within litters. Proteomics studies show that protein synthesis and degradation is downregulated in IUGR pigs and low birth weight pigs also produce meat with less tenderness. Alternative maternal feeding strategies to prevent IUGR have been examined. Increasing maternal global nutrition had no beneficial effect on performance and muscle growth traits in several studies. Feeding excess maternal dietary protein also did not influence muscle growth traits whereas moderately decreased maternal dietary protein may decrease muscle fiber number and performance. On the other hand, addition of L-carnitine to the maternal gestation or lactation diet may increase birth and weaning weights or the muscle fiber number, respectively, in low birth weight pig offspring. Finally, promising data have been obtained on reproductive traits in pigs after addition of functional AA, such as arginine and glutamine, to the gestational diet. Although much is known about the biological basis of IUGR, we still need to learn much more about the mode of action before maternal dietary strategies can be developed to prevent IUGR.

Effects of gestation and transition diets, piglet birth weight, and fasting time on depletion of glycogen pools in liver and 3 muscles of newborn piglets.

The experiment was conducted to assess the effects of maternal nutrition in late gestation on glycogen pools of newborn piglets of different birth weights and to assess how rapidly the glycogen pools in the liver and 3 muscles are mobilized during fasting. Until d 108 of gestation, 48 sows were fed a gestation standard diet (GSD) with low dietary fiber (DF, 17.1%), or 1 of 3 diets with high DF (32.3 to 40.4%) consisting of pectin residue (GPR), potato pulp (GPP), or sugar-beet pulp (GSP). From d 108 until farrowing, sows were fed 1 of 6 transition diets with low or high dietary fat: one group received a standard diet (TSD; control) containing 3% animal fat, another group received the TSD diet + 2.5 g/d of hydroxy-methyl butyrate as topdressing (THB), and 4 other groups received diets with 8% added fat from coconut oil (TCO), sunflower oil (TSO), fish oil (TFO), or 4% octanoic acid + 4% fish oil (TOA). Two piglets per litter (the second and fifth born) were blood sampled, and 1 was killed immediately after birth, whereas the other, depending on the litter, was killed after 12, 24, or 28.5 to 36 h (mean 32.5 h) of fasting. Samples of liver, LM, M. semimembranousus (SM), and M. diaphragm (DP) were collected and analyzed for glycogen concentration. No dietary effects (P > 0.20) on glycogen concentrations in liver, LM, SM, or DP were observed. The weight of the liver was affected by gestation diet (P < 0.05) and was greater in GSD and GSP piglets (36.7 and 36.3 g) than in GPR piglets (32.6 g), and intermediate (33.6 g) in GPP piglets. Liver weight, estimated muscle mass, and glycogen pools (P < 0.001) were affected by birth weight, whereas glycogen concentrations in liver and LM, SM, and DP muscles were not (P > 0.05). Liver weight; glycogen concentrations in liver, LM, SM, and DP; and glycogen pools in liver and muscles decreased (P < 0.001) with increasing duration of fasting, and at 32.5 h of fasting, glycogen concentration was reduced by 80% in liver, 64% in DP, 46% in SM, and 36% in LM. Based on a broken-line model, labile glycogen in SM, a locomotory muscle, was estimated to be depleted after 16.4 h of fasting. In conclusion, piglet size had a major impact on estimated glycogen pools, whereas sow nutrition in late gestation had a minor impact, if any. Furthermore, varying proportions of pools of glycogen present in liver and selected muscles were mobilized, and data indicate that newborn piglets are fatally depleted of energy after 16 h of fasting.

Advances in research on the prenatal development of skeletal muscle in animals in relation to the quality of muscle-based food. I. Regulation of myogenesis and environmental impact.

Skeletal muscle development in vertebrates - also termed myogenesis - is a highly integrated process. Evidence to date indicates that the processes are very similar across mammals, poultry and fish, although the timings of the various steps differ considerably. Myogenesis is regulated by the myogenic regulatory factors and consists of two to three distinct phases when different fibre populations appear. The critical times when myogenesis is prone to hormonal or environmental influences depend largely on the developmental stage. One of the main mechanisms for both genetic and environmental effects on muscle fibre development is via the direct action of the growth hormone-insulin-like growth factor (GH-IGF) axis. In mammals and poultry, postnatal growth and function of muscles relate mainly to the hypertrophy of the fibres formed during myogenesis and to their fibre-type composition in terms of metabolic and contractile properties, whereas in fish hyperplasia still plays a major role. Candidate genes that are important in skeletal muscle development, for instance, encode for IGFs and IGF-binding proteins, myosin heavy chain isoforms, troponin T, myosin light chain and others have been identified. In mammals, nutritional supply in utero affects myogenesis and the GH-IGF axis may have an indirect action through the partitioning of nutrients towards the gravid uterus. Impaired myogenesis resulting in low skeletal myofibre numbers is considered one of the main reasons for negative long-term consequences of intrauterine growth retardation. Severe undernutrition in utero due to natural variation in litter or twin-bearing species or insufficient maternal nutrient supply may impair myogenesis and adversely affect carcass quality later in terms of reduced lean and increased fat deposition in the progeny. On the other hand, increases in maternal feed intake above standard requirement seem to have no beneficial effects on the growth of the progeny with myogenesis not or only slightly affected. Initial studies on low and high maternal protein feeding are published. Although there are only a few studies, first results also reveal an influence of nutrition on skeletal muscle development in fish and poultry. Finally, environmental temperature has been identified as a critical factor for growth and development of skeletal muscle in both fish and poultry.

Advances in research on the prenatal development of skeletal muscle in animals in relation to the quality of muscle-based food. II--Genetic factors related to animal performance and advances in methodology.

Selective breeding is an effective tool to improve livestock. Several selection experiments have been conducted to study direct selection responses as well as correlated responses in traits of skeletal muscle growth and function. Moreover, comparisons of domestic with wild-type species and of extreme breeds provide information on the genetic background of the skeletal muscle phenotype. Structural muscular components that differed with increasing distance in lean growth or meat quality in mammals were found to be myofibre number, myofibre size, proportions of fibre types as well as the numbers and proportions of secondary and primary fibres. Furthermore, markers of satellite cell proliferation, metabolic enzyme activities, glycogen and fat contents, the expression of myosin heavy chain isoforms, of activated AMPKα and other proteins in skeletal muscle tissue and circulating IGF1 and IGF-binding proteins have been identified to be involved in selection responses observed in pigs, cattle and/or chicken. The use of molecular methods for selective breeding of fish has only recently been adopted in aquaculture and studies of the genetic basis of growth and flesh quality traits are scarce. Some of the molecular markers of muscle structure/metabolism in livestock have also been identified in fish, but so far no studies have linked them with selection response. Genome scans have been applied to identify genomic regions exhibiting quantitative trait loci that control traits of interest, for example, muscle structure and meat quality in pigs and growth rate in chicken. As another approach, polymorphisms in candidate genes reveal the relationship between genetic variation and target traits. Thus, in large-scale studies with pigs' associations of polymorphisms in the HMGA2, CA3, EPOR, NME1 and TTN genes with traits of carcass and meat quality were detected. Other studies revealed the significance of mutations in the IGF2 and RYR1 genes for carcass lean and muscle fibre traits in pigs. Mutations in the myostatin (MSTN) gene in fish were also examined. Advances in research of the genetic and environmental control of traits related to meat quality and growth have been made by the application of holistic 'omics' techniques that studied the whole muscle-specific genome, transcriptome and proteome in relation to muscle and meat traits, the development of new methods for muscle fibre typing and the adaptation of biophysical measures to develop parameters of muscle fibre traits as well as the application of in vitro studies. Finally, future research priorities in the field are defined.

Birth weight and postnatal dietary protein level affect performance, muscle metabolism and meat quality in pigs.

Intrauterine growth restriction (IUGR), resulting in low birth body weight (LBW) occurs naturally in pigs. However, IUGR may also cause persistent changes in physiology and metabolism resulting in poorer performance, organogenesis and meat quality. As IUGR pigs have a lower daily gain from birth to slaughter they may differ in utilization of nutrients and requirements for dietary protein compared with their larger littermates. Thus, the objective in this study was to examine the interaction between birth body weight (BW) and the postnatal dietary protein level, in relation to postnatal performance, organogenesis, muscle metabolism and meat quality. The experiment was carried out with offspring from 16 purebred Danish Landrace gilts mated to Danish Landrace boars. The female and entire male pigs with LBW that survived at weaning were compared with the female and male pigs with the highest/high birth body weight (HBW) within each litter. The offspring were reared individually from weaning and were fed ad libitum a diet containing either a normal level of protein (NP) for optimal growth or an isocaloric diet containing a 30% lower protein content (LP) from 3 weeks to 150 days of age. At slaughter, we found no interactions between birth weight group and dietary protein level for any of the measured traits. The relative crown-rump length (cm/kg) at birth indicates that LBW pigs were thinner than HBW pigs. Daily gain and feed intake were reduced by 14% and 10%, respectively, while the kg feed/kg gain was slightly increased by 3% in LBW pigs compared with HBW pigs. The LP diet reduced daily gain by 27% due to reduced feed intake and increased kg feed/kg gain by 12% and 21%, respectively compared with the NP diet. LBW male pigs produced meat with a higher shear force than male HBW pigs and also LP pigs produced meat with higher shear force than NP pigs. The activity of lactate dehydrogenase in the Longissimus dorsi muscle (LD) was reduced in pigs fed the LP diet. Calpastatin was increased in LD of LBW pigs and decreased in pigs fed the NP diet. In conclusion, these results suggest a rejection of our hypothesis that low birth weight littermates have a lower requirement for dietary protein compared with heavy weight littermates. Furthermore, LBW male pigs and LP fed pigs of both genders produced less tender meat than HBW pigs or NP fed pigs, respectively.

Effects of fasting prior to slaughter on pH development and energy metabolism post-mortem in M. longissimus dorsi of pigs.

The aim of this study was to investigate the effect of pre-slaughter fasting time, sex and feeding regime on the development of energy metabolism and pH in M. longissimusdorsi (LD) post-mortem in pigs. Two hundred and seventy pigs of the commercial Norwegian crossbreed Noroc (LYLD) were used involving two sexes (gilts and castrates), two feeding regimes (restricted and ad libitum) and four different fasting treatments: (F4) 4h fasting, (F175) 17.5h fasting on the farm, (FO175) 17.5h fasting overnight at the abattoir, and (FO265) 26.5h fasting overnight at the abattoir. Additionally the pigs experienced two different abattoir lairage times as fasting treatment F4 and F175 had a lairage time of 1.5h, while fasting treatment FO175 and FO265 had a lairage time of 23.0 h. A short fasting time of 4 h led to a delayed degradation of glycogen, slow decline in pH and a lower ultimate pH(45 h) post-mortem (pHu) in the LD compared with a fasting time of 26.5 h which resulted in a rapid breakdown of glycogen and pH decline early post-mortem and a high pHu. Proglycogen was degraded in favour of macroglycogen under anaerobic conditions post-mortem. Feeding the animals in the morning before delivery if slaughtered the same day, results in low pH reduction rate and a low pHu compared with pigs fasted overnight either on farm or at the abattoir. Aiming a higher pHu in LD it should be recommended not to feed the pigs in the morning at the day of slaughter.

NMR-based metabonomics reveals relationship between pre-slaughter exercise stress, the plasma metabolite profile at time of slaughter, and water-holding capacity in pigs.

Nuclear magnetic resonance (NMR)-based metabonomics was applied to investigate the effects of pre-slaughter exercise stress on the plasma metabolite profile at time of slaughter. The study included a total of 40 slaughter pigs, which were exposed to one of the following treatments: No pre-slaughter stress (control treatment), pre-slaughter exercise on a treadmill and subsequently 0, 1, or 3h rest prior to slaughter. NMR-based metabonomics revealed a clear difference in the plasma metabolite profile at time of slaughter between control pigs and pigs exercised without rest, which mainly could be ascribed to increased plasma lactate due to exercise. A resting period of 1 or 3h prior to slaughter reversed the stress-induced perturbations in the plasma metabolite profile. The plasma metabolite profile at time of slaughter was highly correlated with muscle temperature 1 min post-mortem, and a correlation to WHC was also demonstrated. Lactate was found to be the metabolite of importance for the association between the plasma metabolome and pH, temperature and WHC.

In vitro primary satellite cell growth and differentiation within litters of pigs.

Postnatal muscle growth is dependent on satellite cell (SC) proliferation, differentiation and fusion to increase the DNA content of existing muscle fibres and thereby the capacity to synthesize protein. The purpose of the present study was to examine the ability of isolated SCs from low, medium and high weaning weight litter mates of pigs to proliferate and differentiate, and to affect protein synthesis and degradation after fusion into myotubes. At 6 weeks of age, SCs from the lowest weight (LW), medium weight (MW) and highest weight (HW) female pigs within eight litters were isolated. Thereby, eight cultures of SCs were established for each of the three weight groups within litter, representing three groups of SCs from pigs exhibiting differences in postnatal muscle growth performance. Proliferation was estimated as the number of viable cells at different time points after seeding. SC differentiation was evaluated by measuring the activity of the muscle-specific enzyme, creatine phosphokinase, and protein synthesis and degradation were measured by incorporation and release of 3H-tyrosine, respectively. A tendency towards a difference in proliferation between SC cultures was found (P = 0.09). This was evident as the number of viable cells at day 3 was lower in cultures from LW pigs than from HW (P < 0.05) and MW (P < 0.01) pigs. Differentiation was significantly different between cultures (P < 0.05). There was a significant difference between LW and MW cultures at 72 h (P < 0.05), and a tendency towards a difference between LW and HW cultures at 45 h (P = 0.07). Protein synthesis per µg protein or per µg DNA did not differ among SC cultures from LW, MW and HW pigs. Neither did protein degradation rate differ significantly among SC cultures from LW, MW and HW pigs. Overall, the results show that SCs from LW pigs seem to proliferate and differentiate at a slower rate than SCs from MW and HW pigs. The results found in this study show no difference in the ability of SCs to affect protein synthesis or degradation between SCs from litter mates exhibiting different growth rates in vivo.

Rest before slaughter ameliorates pre-slaughter stress-induced increased drip loss but not stress-induced increase in the toughness of pork.

One factor affecting meat quality is pre-slaughter stress. We investigated the effects of exercise stress on drip loss and toughness in relation to resting times of 0, 1 or 3h following exercise on a treadmill. This exercise stress was regarded as combined physical and physiological stress. Exercise stress increased the muscle temperature, reduced the creatine phosphate, ATP and glycogen content of pigs slaughtered immediately after stress exposure. These conditions lead to a reduced pH early post mortem and an increased drip loss, while only 1h of rest after exercise stress normalised these effects. However, an overshooting effect was noted when pigs were rested for 1-3h before slaughter, emphasising the importance of critical control of the resting period when studying exercise stress-induced effects on meat quality. Furthermore, meat from exercise stressed pigs, irrespective of resting, had increased toughness compared to controls, indicating that the toughness was not related to drip loss in meat from exercise stressed pigs.

Effects of fasting prior to slaughter on technological and sensory properties of the loin muscle (M. longissimus dorsi) of pigs.

The aim of this study was to investigate the effect of pre-slaughter fasting time, sex and feeding regime on water-holding capacity (WHC), colour and sensory properties and their relationship with pH in M. longissimusdorsi (LD) in pigs. 270 pigs of the commercial Norwegian crossbreed Noroc (Norwegian Landrace×Yorkshire sow and Norwegian Landrace×Duroc boar) were used involving two sexes (gilts and castrates), two feeding regimes (restricted and ad libitum) and four fasting treatments: (F4) 4h fasting (control), (F175) 17.5h fasting on the farm, (FO175) 17.5h fasting overnight at the abattoir, and (FO265) 26.5h fasting overnight at the abattoir. Additionally, the pigs experienced two abattoir lairage times as fasting treatments F4 and F175had a lairage time of 1.5 h, while fasting treatments FO175 and FO265 had a lairage time of 23.0h. A short fasting time of 4h led to a delayed decline in pH post-mortem and a lower ultimate pH (pHu) in the LD compared with a fasting time of 26.5h which resulted in a rapid pH decline early post-mortem and a high pHu. Prolonged fasting reduced drip loss, resulted in a darker colour and tended to improve tenderness of the LD. Castrates showed lower drip loss, higher lightness and improved tenderness and juiciness compared with gilts, while ad libitum feeding improved tenderness compared to restricted feeding. There are obvious negative relationships between pHu and drip loss, lightness and tenderness of LD.

Temporal changes in glycogenolytic enzyme mRNAs during myogenesis of primary porcine satellite cells.

The objective was to study the regulation of glycogenolytic enzyme mRNAs in porcine satellite cells during proliferation and differentiation. Beyond 80% confluence, cells were grown in absence or presence of 1µM insulin. The observed increases in abundance of mRNA for glycogenin, glycogen synthase, phosphorylase kinase, phosphorylase and glycogen debranching enzyme, and no alterations of the transporter molecule GLUT4, clearly indicate that glycogenolytic enzymes of potential importance to meat quality development are regulated at the gene level during myogenesis, and are heavily involved in muscle cell and muscle fibre development. The genes, however, are not influenced by insulin, and the lack of response to insulin of expression of gene-encoding enzymes involved in the formation and degradation of glycogen may question the applicability of porcine cell culture systems, like the one applied, as a model to study the regulation and regulatory mechanism of energy metabolism in muscles.

Novel method for determination of myofibril fragmentation post-mortem.

Multi angle light scattering was used to determine the myofibril fragmentation of pig longissimus dorsi muscle which was then compared with results from the common turbidity method. The method is based on measurement of the myofibril particle size distribution with the use of a special optical unit containing several individual detectors. The method was able to determine post-mortem changes in a pig muscle homogenate without purification of the myofibrils and is therefore simpler and much faster than the traditional turbidity method. There was a significant correlation (p<0.01) between Warner-Bratzler shear force (WBSF) and particle size distribution. The root mean square error of prediction was found to be 6.1N (10-15% of the measured WBSF) when multivariate data analysis was used to make a prediction model for WBSF. Multi angle light scattering is very useful for estimation of myofibril fragmentation since the method is fast and the sample preparation is simple.

In vitro and in vivo studies of creatine monohydrate supplementation to Duroc and Landrace pigs.

Duroc and Landrace pigs as well as primary myotubes from these breeds were used to investigate mechanisms behind differences in their response to creatine monohydrate (CMH). Pigs were supplemented with 0, 12.5, 25 or 50g CMH/d for 5 days (n=10 per treatment and breed). Plasma levels of creatine increased dose-dependently in both breeds, while muscle-creatine phosphate content increased only in the Duroc pigs. (1)H NMR metabolic profiling showed a tendency towards clustering according to CMH supplementation only among Duroc pigs, revealing a stronger response compared to Landrace pigs. The abundance of insulin-like growth factor I and myostatin mRNA was decreased by CMH supplementation while that of type 1 IGF-receptor and creatine transporter was unaffected. Protein synthesis, increased in the myotubes from both breeds, indicating protein accretion, but no effect was observed on the mRNA abundance of IGF-I, type 1 IGF-receptor, myostatin or the creatine transporter in myotubes.