5`-aminoimidazole-4-carboxyamide-ribonucleoside- activated glucose transport is not prevented by nitric oxide synthase inhibition in rat isolated skeletal muscle

1. The nucleoside intermediate 5'-aminoimidazole-4-carboxyamide-ribonucleoside (AICAR) activates skeletal muscle AMP-activated protein kinase (AMPK) and increases glucose uptake. The AMPK phosphorylates neuronal nitric oxide synthase (nNOS)µ in skeletal muscle fibres. There is evidence that both AMPK and nNOSµ may be involved in the regulation of contraction-stimulated glucose uptake.
2. We examined whether both AICAR- and contraction-stimulated glucose uptake were mediated by NOS in rat skeletal muscle.
3. Rat isolated epitrochlearis muscles were subjected in vitro to electrically stimulated contractions for 10 min and/or incubated in the presence or absence of AICAR (2 mmol/L) or the NOS inhibitor NG-monomethyl-l-arginine (l-NMMA; 100 µmol/L).
4. Muscle contraction significantly (P < 0.05) altered the metabolic profile of the muscle. In contrast, AICAR and l-NMMA had no effect on the metabolic profile of the muscle, except that AICAR increased muscle 5'-aminoimidazole-4-carboxyamide-ribonucleotide (ZMP) and AICAR content. Nitric oxide synthase inhibition caused a small but significant (P < 0.05) reduction in basal 3-O-methylglucose transport, which was observed in all treatments. 5'-Aminoimidazole-4-carboxyamide-ribonucleoside significantly increased (P < 0.05) glucose transport above basal, with NOS inhibition decreasing this slightly (increased by 209% above basal compared with 184% above basal with NOS inhibition). Contraction significantly increased glucose transport above basal, with NOS inhibition substantially reducing this (107% increase vs 31% increase). 5'-Aminoimidazole-4-carboxyamide-ribonucleoside plus contraction in combination were not additive on glucose transport.
5. These results suggest that NO plays a role in basal glucose uptake and may regulate contraction-stimulated glucose uptake. However, NOS/nitric oxide do not appear to be signalling intermediates in AICAR-stimulated skeletal muscle glucose uptake.

Alterations in the proportions of skeletal muscle proteins following a unilateral lesion to the substantia nigra pars compacta of rats

It is well established that mammalian skeletal muscles exhibit a considerable degree of plasticity and one of the main determining factors of this plasticity is the activity pattern and duration of motoneurone discharge. Lesions to the right substantia nigra pars compacta (SNpc) of six adult rats were made to determine whether altered output from the SNpc ultimately leads to a change in the expression of proteins in contralateral skeletal muscles. After 4 months, altered motor performance was identified by the administration of amphetamine. After 7 months, 30–70% of dopaminergic cells in the SNpc had been destroyed. The protein content of muscles was then quantified from densitometric scans of gels, and expressed as a % of the amount of actin (the protein used as a reference in this study). The lesion affected the expression of different protein isoforms in the fast- and slow-twitch muscles. In slow-twitch soleus muscles, the lesion decreased the proportion of α-tropomyosin and increased the proportion of β-tropomyosin. In the fast-twitch extensor digitorum longus muscles, the lesion increased the proportion of the fast isoform of troponin-T_1f, and decreased the proportions of the two isoforms of myosin light chain. This study establishes a connection between the chronic effects of a lesion to the SNpc, with a loss of dopaminergic neurones, impaired motor performance, and altered expression of proteins in skeletal muscle. The implication of these results is that the altered motor function observed in Parkinson’s disease may be associated with alterations to the expression of skeletal muscle proteins.

Differential effects of arginine, glutamate and phosphoarginine on Ca2+-activation properties of muscle fibres from crayfish and rat

The effects of two amino acids, arginine which has a positively charged side-chain and glutamate which has a negatively charged side-chain on the Ca²⁺-activation properties of the contractile apparatus were examined in four structurally and functionally different types of skeletal muscle; long- and short-sarcomere fibres from the claw muscle of the yabby (a freshwater decapod crustacean), and fast- and slow-twitch fibres from limb muscles of the rat. Single skinned fibres were activated in carefully balanced solutions of different pCa (-log₁₀[Ca²⁺]) that either contained the test solute (“test”) or not (“control”). The effect of phosphoarginine, a phosphagen that bears a nett negative charge, was also compared to the effects of arginine. Results show that (i) arginine (33-36 mmol l^-1) significantly shifted the force–pCa curve by 0.08–0.13 pCa units in the direction of increased sensitivity to Ca²⁺-activated contraction in all fibre types; (ii) phosphoarginine (9–10 mmol l^-1) induced a significant shift of the force–pCa curve by 0.18–0.24 pCa units in the direction of increased sensitivity to Ca²⁺ in mammalian fast- and slow-twitch fibres, but had no significant effects on the force–pCa relation in either long- or short-sarcomere crustacean fibres; (iii) glutamate (36–40 mmol l^-1), like arginine affected the force–pCa relation of all fibre types investigated, but in the opposite direction, causing a significant decrease in the sensitivity to Ca²⁺-activated contraction by 0.08–0.19 pCa units; (iv) arginine, phosphoarginine and glutamate had little or no effect on the maximum Ca²⁺-activated force of crustacean and mammalian fibres. The results suggest that the opposing effects of glutamate and arginine are not related to simply their charge structure, but must involve complex interactions between these molecules, Ca²⁺ and the regulatory and other myofibrillar proteins.

Properties of slow- and fast-twitch muscle fibres in a mouse model of amyotrophic lateral sclerosis

This investigation was undertaken to determine if there are altered histological, pathological and contractile properties in presymptomatic or endstage diseased muscle fibres from representative slow-twitch and fast-twitch muscles of SOD1 G93A mice in comparison to wildtype mice. In presymptomatic SOD1 G93A mice, there was no detectable peripheral dysfunction, providing evidence that muscle pathology is secondary to motor neuronal dysfunction. At disease endstage however, single muscle fibre contractile analysis demonstrated that fast-twitch muscle fibres and neuromuscular junctions are preferentially affected by amyotrophic lateral sclerosis-induced denervation, being unable to produce the same levels of force when activated by calcium as muscle fibres from their age-matched controls. The levels of transgenic SOD1 expression, aggregation state and activity were also examined in these muscles but there no was no preference for muscle fibre type. Hence, there is no simple correlation between SOD1 protein expression/activity, and muscle fibre type vulnerability in SOD1 G93A mice.

Age and walking speed effects on muscle recruitment in gait termination

During gait termination at normal walking speed, older adults more frequently employ two-step responses, increasing their stopping distance and stopping time more than younger controls. This study investigated ageing effects on lower limb muscle recruitment patterns during stopping at three walking speeds. Twelve young male (26±3.7 years, range 19–30) and 12 gender-matched older participants (72±4.3 years, range 65–82) terminated walking at normal, medium and maximum speed. A visual stopping stimulus was presented 10 ms following either left or right heel-contact with no stimulus (catch) on 30% of trials. Electromyographic (EMG) activity was recorded from the tibialis anterior (TA), soleus (SOL), biceps femoris (BF), vastus lateralis (VL) and gluteus medius (GM). Older males more frequently (46% of trials) took two-steps to stop than young males (20%). The stance leg muscles responded significantly faster than the swing leg, and with increased speed, fewer swing limb muscles contributed to stopping. Older males were slower to respond with the stance leg, at 215 ms following the stimulus compared with 176 ms for the younger group. They also recruited fewer swing leg muscles with less frequent activation of the soleus and gluteus medius. Failure to activate muscles would provide less extensor torque to maintain the centre of gravity anterior to the forward base of support. This would decrease the total force opposing horizontal velocity in order to bring the body to rest and, as a consequence, encourage an additional step prior to stopping.

µ- Calpain and calpain-3 are not autolyzed with exhaustive exercise in humans

µ-calpain and calpain-3 are Ca²⁺-dependent proteases found in skeletal muscle. Autolysis of calpains is observed using Western blot analysis as the cleaving of the full-length proteins to shorter products. Biochemical assays suggest that µ-calpain becomes proteolytically active in the presence of 2–200 µM Ca²⁺. Although calpain-3 is poorly understood, autolysis is thought to result in its activation, which is widely thought to occur at lower intracellular Ca²⁺ concentration levels ([Ca²⁺]i; ~1 µM) than the levels at which µ-calpain activation occurs. We have demonstrated the Ca²⁺-dependent autolysis of the calpains in human muscle samples and rat extensor digitorum longus (EDL) muscles homogenized in solutions mimicking the intracellular environment at various [Ca²⁺] levels (0, 2.5, 10, and 25 µM). Autolysis of calpain-3 was found to occur across a [Ca²⁺] range similar to that for µ-calpain, and both calpains displayed a seemingly higher Ca²⁺ sensitivity in human than in rat muscle homogenates, with ~15% autolysis observed after 1-min exposure to 2.5 µM Ca²⁺ in human muscle and almost none after 1- to 2-min exposure to the same [Ca²⁺]i level in rat muscle. During muscle activity, [Ca²⁺]i may transiently peak in the range found to autolyze µ-calpain and calpain-3, so we examined the effect of two types of exhaustive cycling exercise (30-s "all-out" cycling, n = 8; and 70% VO2 peak until fatigue, n = 3) on the amount of autolyzed µ-calpain or calpain-3 in human muscle. No significant autolysis of µ-calpain or calpain-3 occurred as a result of the exercise. These findings have shown that the time- and concentration-dependent changes in [Ca²⁺]i that occurred during concentric exercise fall near but below the level necessary to cause autolysis of calpains in vivo.

Effect of AICAR treatment on gycogen metabolism in skeletal muscle

AMP-activated protein kinase (AMPK) is proposed to stimulate fat and carbohydrate catabolism to maintain cellular energy status. Recent studies demonstrate that pharmacologic activation of AMPK and mutations in the enzyme are associated with elevated muscle glycogen content in vivo. Our purpose was to determine the mechanism for increased muscle glycogen associated with AMPK activity in vivo. AMPK activity and glycogen metabolism were studied in red and white gastrocnemius muscles from rats treated with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) in vivo, and also in muscles incubated with AICAR in vitro. In vivo AICAR treatment reduced blood glucose and increased blood lactate compared with basal values. AICAR increased muscle α2 AMPK activity, glycogen, and glucose-6-phosphate concentrations. Glycogen synthase activity was increased in the red gastrocnemius but was decreased in the white gastrocnemius. Glycogen phosphorylase activity increased in both muscles, with an inhibition initially observed in the red gastrocnemius. In vitro incubation with AICAR activated α2 AMPK but had no effect on either glycogen synthase or glycogen phosphorylase. These results suggest that AICAR treatment does not promote glycogen accumulation in skeletal muscle in vivo by altering glycogen synthase and glycogen phosphorylase. Rather, the increased glycogen is due to the well-known effects of AICAR to increase glucose uptake.

Expression of uncoupling protein-3 subsarcolemmal and intermyofibrillar mitochondria of various mouse muscle types and its moduclation by fasting

Uncoupling protein-3 (UCP3) is a mitochondrial inner-membrane protein abundantly expressed in rodent and human skeletal muscle which may be involved in energy dissipation. Many studies have been performed on the metabolic regulation of UCP3 mRNA level, but little is known about UCP3 expression at the protein level. Two populations of mitochondria have been described in skeletal muscle, subsarcolemmal (SS) and intermyofibrillar (IMF), which differ in their intracellular localization and possibly also their metabolic role. To examine if UCP3 is differentially expressed in these two populations and in different mouse muscle types, we developed a new protocol for isolation of SS and IMF mitochondria and carefully validated a new UCP3 antibody. The data show that the density of UCP3 is higher in the mitochondria of glycolytic muscles (tibialis anterior and gastrocnemius) than in those of oxidative muscle (soleus). They also show that SS mitochondria contain more UCP3 per mg of protein than IMF mitochondria. Taken together, these results suggest that oxidative muscle and the mitochondria most closely associated with myofibrils are most efficient at producing ATP. We then determined the effect of a 24-h fast, which greatly increases UCP3 mRNA (16.4-fold) in muscle, on UCP3 protein expression in gastrocnemius mitochondria. We found that fasting moderately increases (1.5-fold) or does not change UCP3 protein in gastrocnemius SS or IMF mitochondria, respectively. These results show that modulation of UCP3 expression at the mRNA level does not necessarily result in similar changes at the protein level and indicate that UCP3 density in SS and IMF mitochondria can be differently affected by metabolic changes.

Redistribution of glucose from skeletal muscle to adipose tissue during catch-up fat. A link between catch-up growth and later metabolic syndrome.

Catch-up growth, a risk factor for later obesity, type 2 diabetes, and cardiovascular diseases, is characterized by hyperinsulinemia and an accelerated rate for recovering fat mass, i.e., catch-up fat. To identify potential mechanisms in the link between hyperinsulinemia and catch-up fat during catch-up growth, we studied the in vivo action of insulin on glucose utilization in skeletal muscle and adipose tissue in a previously described rat model of weight recovery exhibiting catch-up fat caused by suppressed thermogenesis per se. To do this, we used euglycemic-hyperinsulinemic clamps associated with the labeled 2-deoxy-glucose technique. After 1 week of isocaloric refeeding, when body fat, circulating free fatty acids, or intramyocellular lipids in refed animals had not yet exceeded those of controls, insulin-stimulated glucose utilization in refed animals was lower in skeletal muscles (by 20–43%) but higher in white adipose tissues (by two- to threefold). Furthermore, fatty acid synthase activity was higher in adipose tissues from refed animals than from fed controls. These results suggest that suppressed thermogenesis for the purpose of sparing glucose for catch-up fat, via the coordinated induction of skeletal muscle insulin resistance and adipose tissue insulin hyperresponsiveness, might be a central event in the link between catch-up growth, hyperinsulinemia and risks for later metabolic syndrome.

Skeletal muscle htererogeneity in fasting-induced upregulation of genes encoding UCP2, UCP3, PPARĂ and key enzymes of lipid oxidation

The uncoupling protein homologs UCP2 and UCP3 have been proposed as candidate genes for the regulation of lipid metabolism. Within the context of this hypothesis, we have compared, from fed and fasted rats, changes in gene expression of skeletal muscle UCP2 and UCP3 with those of carnitine palmitoyltransferase I and medium-chain acyl-CoA dehydrogenase, two key enzymes regulating lipid flux across the mitochondrial #-oxidation pathway. In addition, changes in gene expression of peroxisome proliferator-activated receptor gamma, a nuclear transcription factor implicated in lipid metabolism, were also investigated. The results indicate that in response to fasting, the mRNA levels of UCP2, UCP3, carnitine palmitoyltransferase I and medium-chain acyl-CoA dehydrogenase are markedly increased, by three- to sevenfold, in the gastrocnemius and tibialis anterior (fast-twitch muscles, predominantly glycolytic or oxidative-glycolytic), but only mildly increased, by less than twofold, in the soleus (slow-twitch muscle, predominantly oxidative). Furthermore, such muscle-type dependency in fasting-induced transcriptional changes in UCP2, UCP3, carnitine palmitoyltransferase and medium-chain acyl-CoA dehydrogenase persists when the increase in circulating levels of free fatty acids during fasting is abolished by the anti-lipolytic agent nicotinic acid - with blunted responses only in the slow-twitch muscle contrasting with unabated increases in fast-twitch muscles. Independently of muscle type, however, the mRNA levels of peroxisome proliferator-activated receptor gamma are not altered during fasting. Taken together, these studies indicate a close association between fasting-induced changes in UCP2 and UCP3 gene expression with those of key regulators of lipid oxidation, and are hence consistent with the hypothesis that these UCP homologs may be involved in the regulation of lipid metabolism. Furthermore, they suggest that in response to fasting, neither the surge of free fatty acids in the circulation nor induction of the peroxisome proliferator-activated receptor gamma gene may be required for the marked upregulation of genes encoding the UCP homologs and key enzymes regulating lipid oxidation in fast-twitch muscles.

Effect of dietary modificatio of muscle long-chainN-3 fatty acid on plasma insulin and lipid metabolite, carcass traits, and fat deposition in lambs

In a previous study we showed that feeding fish meal significantly increased muscle long chain n-3 fatty acids (FA) and hot carcass weight. In this study we compared the effect of fish meal and fish oil on increasing muscle long-chain FA. We also investigated whether the increase in carcass weight was due to the effect of dietary enrichment of muscle long-chain n-3 FA on muscle membrane phospholipids and(or) to rumen by-pass protein provided by fish meal. Forty crossbred ([Merino x Border Leicester] x Poll Dorset) wether lambs between 26 and 33 kg BW were randomly assigned to one of five treatments: 1) basal diet of oaten:lucerne chaff (Basal); 2) Basal + fish meal (9% DM) = FM; 3) Basal + fish oil (1.5% DM) with protected sunflower meal (9% DM ) = FOSMP; 4) Basal + fish oil (1.5% DM) = FO; or 5) Basal + protected sunflower meal (10.5% DM) = SMP. Daily intake of ME (9.60 - 10.5 MJ ME/d) and CP (150 to 168 g/d) in all treatments was kept similar by varying the ratio of oaten:lucerne chaff and by feeding the animals at 90% ad libitum intake. Blood samples were collected at the start of the experiment and on the day (d 42) prior to slaughter. Lambs were then slaughtered at a commercial abattoir. At 24 h postmortem carcass traits were measured and longis-simus thoracis muscle taken for analysis of FA of phospholipid and triglyceride fractions. Lambs fed FO and FOSMP showed a marked increase in muscle longchain n-3 FA (P < 0.001) and a reduction in magnitude of the rise in insulin concentration (P < 0.001) after feeding compared with lambs fed Basal and SMP diets. Lambs in FM had a moderate increase (P < 0.001) in muscle long-chain n-3 FA content. Compared with Basal diet, both plasma total cholesterol (P < 0.02) and high-density lipoprotein cholesterol (P < 0.001) levels were greater in SMP and less in FO and FOSMP treat- ments. The i.m. fat content was reduced (P < 0.05) in FM and FO treatments, but carcass weight was increased only with fish meal (P < 0.03). Adding SMP to FO produced muscle with an intermediate level of i.m. fat, whereas muscle long-chain n-3 FA, i.m. fat, and insulin concentration were unchanged with SMP treatment. These results indicate that an increase in carcass weight in FM may be due to the supply of ruminally undegraded protein. They also suggest that fish oil along with fish meal can increase long-chain n-3 FA content in phospholipid of muscle membrane. This may be associated with reduced i.m. fat content and altered insulin action and lipoprotein metabolism.

Effect of diets containing n-3 fatty acids on muscle long-chain n-3 fatty acid content in lambs fed low- and medium-quality roughage diets

In two experiments, each with 32 crossbred ([Merino x Border Leicester] x Poll Dorset) wether lambs (26 to 33 kg weight range), animals were randomly assigned to one of four treatments. A mixture of lucerne chaff:oaten chaff was used as a basal diet, offered in different ratios. Animals were allowed to consume on a free-access basis in Exp. 1 or 90% of ad libitum intake in Exp. 2 in order to provide a low- (6.5 MJ ME/d) and medium- (9.5 MJ ME/d) quality basal diet, respectively. Isoenergetic amounts of lipid supplements, fish meal (80 g DM), canola meal (84 g DM), and soy meal (75 g DM) were tested in Exp. 1. In Exp. 2, fish meal (9% DM), unprotected rapeseed (7% DM), and protected canola seed (6% DM) were fed as supplements. At the end of 53-d (Exp. 1) or 46-d (Exp. 2) experimental periods, lambs were slaughtered at a commercial abattoir and at 24 h postmortem longissimus thoracis (LT) muscle was collected for the analysis of fatty acid (FA) composition of structural phospholipid and storage triglyceride fractions. Fish meal diet increased LT muscle long-chain n-3 FA content by 27% (P < 0.02) in Exp. I and 30% (P < 0.001) in Exp. 2 compared with lambs fed the basal diet, but fish meal decreased (P < 0.01) the n-6 FA content only in Exp. 1. Soy meal and protected canola seed diets increased (P < 0.01) LT muscle n-6 FA content but did not affect long-chain n-3 FA content. Longissimus thoracis muscle long-chain n-3 FA were mainly deposited in structural phospholipid, rather than in storage triglyceride. In both Exp. 1 and Exp. 2, the ratio of n-6:n-3 FA in LT muscle was lowest (P < 0.01) in lambs fed fish meal supplement compared with all other treatments. Protected canola seed diet increased the ratio of n-6:n-3 FA (P < 0.01) and PUFA:saturated fatty acid (P < 0.03) content from those animals fed the basal, fish meal, and unprotected rapeseed diets in Exp. 2. This was due to an increase in muscle n-6 FA content, mainly linoleic acid, of both phospholipid (P < 0.001) and triglyceride (P < 0.01) fractions and not to an increase in muscle n3 FA content. The results indicate that by feeding fish meal supplement, the essential n-3 FA can be increased while lowering the ratio of n-6:n-3 content in lamb meat to an extent that could affect nutritional value, attractiveness, and the economic value of meat.

Dietary manipulation of muscle long-chain omega-3 and omega-6 fatty acids and sensory properties of lamb meat

The effects of dietary manipulation of muscle long-chain omega-3 fatty acids (FA) on sensory properties of cooked meat in second cross ([Merino×Border Leicester]×Poll Dorset) wether lambs were evaluated. Lambs fed dietary supplements of fish meal (FM, Exp. 1) and fish oil (FO, Exp. 2) showed moderately (P<0.01) and markedly (P<0.001) increased muscle long-chain omega-3 FA content compared with those fed the basal diet of lucerne chaff and oat chaff. Protected canola seed (PCS, Exp. 1) significantly (P<0.001) increased omega-6 FA content of the longissimus muscle. In each of the 2 experiments (1 and 2), after being fed experimental diets for 6 weeks lambs were slaughtered at a commercial abattoir. At 24 h post-mortem (PM) the semitendinosus and biceps femoris muscles were removed from animals and stored at −20°C until evaluation of sensory properties using experienced panel members. The muscle samples were stored for 3 (Exp. 1) and 12 (Exp. 2) months then removed, thawed and cooked for sensory evaluation. The meat samples were cooked under standardized conditions in a convection microwave at 180°C (20–25 min) to an internal temperature of 75°C. Cooked samples were tested for flavour, aroma, juiciness and overall palatability. The significant increase in muscle long-chain omega-3 with FM (Exp. 1 and 2) and FO (Exp. 2) or omega-6 FA with PCS (Exp. 1) were not detrimental to sensory panel evaluations of flavour or aroma of cooked meat when compared with the basal diet. However, meat from FM (Exp. 1) had lower juiciness and FO (Exp. 2) had lower overall palatability. Protected sunflower meal protein with FO (Exp. 2) significantly lowered ratings for flavour, juiciness and overall palatability. Lamb meat with increased levels of long-chain omega-3 FA can be produced without altering the sensory quality (flavour or aroma) of the cooked meat.

Effects of dietary lipid type on muscle fatty acid composition, carcass leanness, and meat toughness in lambs.

Isonitrogenous amounts of two protein sources differing in rumen degradation rate and in lipid composition were fed to sheep with or without a rapidly fermentable cereal grain. The effects on intake, carcass leanness, and muscle fatty acid (FA) composition were examined. Thirty-eight crossbred wether lambs (9 mo, 35 to 48 kg) were allocated by stratified randomization to six treatment groups: 1) basal diet of alfalfa hay:oat hay (20:80) ad libitum = basal; 2) basal + lupin (358 g DM/d) = lupin; 3) basal + fish meal (168 g DM/d) = fish meal; 4) basal + barley (358 g DM/d) = barley; 5) basal + barley + lupin (179 + 179 g DM/d) = barley/lupin; or 6) basal + barley + fish meal (179 + 84 g DM/d) = barley/ fish meal. Lambs were fed individually. Dietary treatments were imposed for 8 wk, and the supplements were offered at 2-d intervals. Daily feed intake and weekly BW of lambs were recorded. At the end of the feeding period lambs were slaughtered after an overnight fast. Hot carcass weight (HCW) and fat depth (GR; total fat and muscle tissue depth at 12th rib, 110 mm from midline) were recorded. At 24 h postmortem samples of longissimus thoracis (LT) and longissimus lumborum (LL) muscles were taken from chilled (4 deg C) carcasses for the assessment of FA composition and meat tenderness, respectively. Lambs fed lupin or fish meal with or without barley had heavier slaughter weights (P < 0.004) and HCW (P < 0.001) than lambs fed basal or barley when initial BW was included as a covariate. The lupin diet also resulted in heavier carcasses (P < 0.05) than the fish meal or barley/fish meal diets. With GR as an indicator, fish meal and barley/ fish meal diets produced leaner carcasses (P < 0.01) than lupin and barley/lupin lambs. Long-chain n-3 FA content [20:5n-3 (P < 0.001), 22:5n-3 (P < 0.003), and 22:6n-3 (P < 0.001)] in the LT muscle were substantially higher with the fish meal and barley/fish meal diets, whereas muscle total n-6 FA was increased (P < 0.003) by lupin and barley/lupin compared with all other diets. Thus, increased muscle long-chain n-3 FA content occurred without an increase in fatness measured as GR, whereas increased muscle n-6 FA content was associated with an increase in carcass fatness. Under these circumstances, a reduction in carcass fatness had no effect on meat tenderness measured as Warner-Bratzler shear force.

Constructing artificial images of facial expressions

This paper presents a method for construction of artificial images of facial expressions. The proposed fractal-based synthesis procedure called pixel-based correspondence works on 2D images and does not require any depth information. This method can generate artificial images of an object when only a single image is given. Using the proposed method, effective example-based facial analysis systems can be trained and utilised in various applications.