Foreleg Muscles Research Articles

Supplementation with a phenolic-rich fraction from Ilex kudingcha improves inflammatory stress, gastrocnemius weights, and foreleg muscle grip strength in murine squamous cell carcinoma (SCC7)-bearing mice

Ethanol supernatant of Ilex kudingcha methanol extract (ESIKME) was identified to be rich in phenolic compounds and inhibited murine squamous cell carcinoma (SCC)-7 growth through slightly restoring adaptive immunity and Th1-inclination immune balance in the SCC7-bearing mice. To further evaluate the inhibitory effects and mechanisms of ESIKME against SCC7-induced inflammation status, the effects of ESIKME on inflammatory stress and related cachexia induced by SCC7 cells were investigated in vitro and in vivo. Chemical components of ESIKME were measured using an advanced LC-MS/MS analysis. SCC7 cells were treated with mouse splenocyte (SCM) and macrophage conditioned media (MCM) cultured without/with ESIKME in vitro. SCC7-bearing mice were orally administered with ESIKME for 26 days in vivo. Cachexia-related biomarkers in the SCC7-bearing mice were analyzed. Most of the detected compounds in ESIKME were polymeric or oxidized phenolic compounds. Either SCM or MCM without ESIKME significantly inhibited SCC7 cell growth. The high-dose ESIKME-administered group (400 mg/kg b.w./day) had significantly higher body weights, feed efficiency, and energy efficiency than the dietary control (DC) group, suggesting non-adverse toxic effects for ESIKME supplementation in the SCC7-bearing mice. The DC group had a higher TNF-α and TNF-α/IL-10 secretion ratio but lower anti-inflammatory IL-10 cytokine secretions by macrophages than the vehicle control group. ESIKME administration dose-dependently and significantly reversed these adverse statuses. Medium dose ESIKME (100 mg/kg b.w./day) decreased tumor sizes but increased gastrocnemius weights and foreleg muscle grip strength. Our results suggest that ESIKME alleviated inflammatory stress and related cachexia in vitro and in vivo.

Reprint of: The European badger, Meles Meles, as a new host for Trichinella britovi in Romania

The European badger, Meles meles (Carnivora, Mustelidae) is a widespread opportunistic omnivorous mammal. Its food spectrum comprises a wide variety of plants and small animals, occasionally including rodents, such as mice or rats. Considering that rodents are known to play a key role in the life cycle of Trichinella spp., the aims of this study were to investigate the occurrence of these parasites in badgers from Romania and to identify the species. Overall, 61 badgers originating from 14 counties were examined by trichinoscopy and artificial digestion. For species determination, the positive muscle samples, and the larvae recovered from the artificial digestion were used for DNA isolation, and further processed by multiplex PCR. A single badger, originating from Sibiu County, Central Romania, was found positive for Trichinella spp. Five cysts were identified using trichinoscopy: four in the diaphragm and one in the foreleg muscles. Artificial digestion revealed an infection rate of 70 larvae/100 g of muscle. The PCR indicated the occurrence of T. britovi, which is the most commonly detected species in wild carnivores in temperate areas. Although T. britovi has previously been reported in Romania, this represents the first report of its occurrence in the European badger in the country. However, the low prevalence indicates a minor reservoir role of this species.

Comparison of Direct and Indirect Toxoplasma gondii Detection and Genotyping in Game: Relationship and Challenges.

The importance of game as a source of Toxoplasma gondii (T. gondii) infection in humans is largely unknown. New data on the presence of T. gondii in game hunted in the Federal State of Brandenburg, Germany, were obtained by direct and indirect detection (ELISA). DNA extracted either directly (5 g heart or foreleg muscle, DE) or after acid pepsin digestion (50 g heart, PD) or enriched by magnetic capture (50 g heart, MC) was examined by real-time PCR (qPCR). ELISA revealed seroprevalences of 20% in wild boar (Sus scrofa), 11% in roe deer (Capreolus capreolus) and 6% in red deer (Cervus elaphus). T. gondii DNA was detected by at least one direct detection method in 12% of wild boar, 6% of roe deer, 2% of fallow deer (Dama dama) and 2% of red deer. In both, positive wild boar and roe deer, T. gondii type II specific alleles were the most prevalent, as assessed by PCR-restriction fragment length polymorphism. The highest proportion of positive animals was detected by MC qPCR, followed by PD qPCR with a similar proportion of positive findings. Investigation of 50 g of heart muscle revealed a significantly higher proportion of positive qPCR results than analysis of 5 g (p = 0.048). An association between seropositivity and direct detection was evident in wild boar and roe deer (p < 0.001). Infectivity of T. gondii DNA–positive samples was confirmed by bioassay (4/4), providing evidence that game could represent a relevant source of viable T. gondii posing a risk for human infection.

The European badger, Meles meles, as a new host for Trichinella britovi in Romania

The European badger, Meles meles (Carnivora, Mustelidae) is a widespread opportunistic omnivorous mammal. Its food spectrum comprises a wide variety of plants and small animals, occasionally including rodents, such as mice or rats. Considering that rodents are known to play a key role in the life cycle of Trichinella spp., the aims of this study were to investigate the occurrence of these parasites in badgers from Romania and to identify the species. Overall, 61 badgers originating from 14 counties were examined by trichinoscopy and artificial digestion. For species determination, the positive muscle samples, and the larvae recovered from the artificial digestion were used for DNA isolation, and further processed by multiplex PCR. A single badger, originating from Sibiu County, Central Romania, was found positive for Trichinella spp. Five cysts were identified using trichinoscopy: four in the diaphragm and one in the foreleg muscles. Artificial digestion revealed an infection rate of 70 larvae/100 g of muscle. The PCR indicated the occurrence of T. britovi, which is the most commonly detected species in wild carnivores in temperate areas. Although T. britovi has previously been reported in Romania, this represents the first report of its occurrence in the European badger in the country. However, the low prevalence indicates a minor reservoir role of this species.

Effects of dietary vitamin B6 on the skeletal muscle protein metabolism of growing rabbits

This study aimed to evaluate the effects of dietary vitamin B6 on the skeletal muscle protein metabolism and expression of transcription and growth factor of growing rabbits. Two hundred, healthy, rabbits with similar bodyweights were randomly assigned to one of five dietary groups with 40 animals per group. The dietary groups consisted of the following different vitamin B6 supplementation levels: 0, 5, 10, 20 and 40 mg/kg. The feeding trial lasted 60 days. The results showed that dietary vitamin B6 elicited significant effects on the fore and hind leg muscle ratio (the fore and hind leg muscle weight/the liveweight; P &lt; 0.05) and on serum total amino acids (T-AA), blood urea and insulin-like growth factor 1 (IGF1) content (P &lt; 0.05). Additionally, expression of IGF1, myogenic determination factor (MYOD) and myogenin (MYOG), myocyte regulation factor 5 (MYF5), myostatin (MSTN) and WW domain-containing E3 proteasome ubiquitin ligase 1 (WWP1) mRNA in the loin (M. longissimus dorsi) were affected by vitamin B6 in diets (P &lt; 0.05). The immunoblot analysis revealed that dietary vitamin B6 elicited significant effects on IGF1, MYOG and WWP1 expression in the loin (P &lt; 0.05). Our results indicate that the addition of dietary vitamin B6 can significantly alter the protein metabolism of growing rabbits and that an appropriate vitamin B6 supplementation level is 20 mg/kg for 3–5-month-old growing rabbits (the basic diet vitamin B6 content was 4.51 mg/kg).

Emerging Trichinella britovi infections in free ranging pigs of Greece

Trichinella infections in humans and pigs have been documented in Greece since 1945 and a high prevalence of infection in pigs occurred in the 1950s. Up to 1984 only sporadic infections in humans were documented, and this zoonosis was not considered as a public health problem until 2009 when a human outbreak caused by the consumption of pork from an organic pig farm occurred. In the present study, we describe the re-emergence of Trichinella spp. infections in free-ranging pigs from organic farms of 3 counties (Dramas, Evros and Kavala) in Northern-Eastern Greece during the period 2009–2012. Totally 37 out of 12,717 (0.29%) free-ranging pigs which were tested during the period in question, were positive for Trichinella spp. larvae. The etiological agent was identified as Trichinella britovi. The average larval burden was 13.7 in the masseter, 6.2 in the foreleg muscles and 7.5 in the diaphragm. The 37 positive animals originated from seven free range pig farms. The practice of organic pig production systems in Greece has grown in popularity over the last years due to the increasing interest of consumers for products considered as traditional. However, this type of pig production increases the risk for Trichinella spp. infections, since animals can acquire the infection by feeding on carcasses or the offal of hunted or dead wild animals. The awareness and education of hunters and farmers is extremely important to reduce the transmission among free ranging pigs and the risk for humans.

Transsynaptic inhibition of spinal transmission by A2 botulinum toxin.

Type A botulinum toxin blocks not only ACh release from motor nerve terminals but also central synaptic transmission, including glutamate, noradrenaline, dopamine, ATP, GABA and glycine. Neurotoxins (NTXs) are transported by both antero- and retrogradely along either motor or sensory axons for bidirectional delivery between peripheral tissues or the CNS. A newly developed type A2 NTX (A2NTX) injected into one rat foreleg muscle was transported to the contralateral muscle. This finding was consistent with the NTX traveling retrogradely via spinal neurons and then transsynaptically through motor neurons to the contralateral motor neurons within the spinal cord and on to the soleus muscle. In the present study we found that toxin injection into the rat left soleus muscle clearly induced bilateral muscle relaxation in a dose-dependent fashion, although the contralateral muscle relaxation followed the complete inhibition of toxin-injected ipsilateral muscles. The toxin-injected ipsilateral muscle relaxation was faster and stronger in A2NTX-treated rats than A1LL (BOTOX). A1LL was transported almost equally to the contralateral muscle via neural pathways and the bloodstream. In contrast, A2NTX was mainly transported to contralateral muscles via the blood. A1LL was more successfully transported to contralateral spinal neurons than A2NTX. We also demonstrated that A1LL and A2NTX were carried from peripheral to CNS and vice versa by dual antero- and retrograde axonal transport through either motor or sensory neurons.

Noninvasive monitoring of treatment response in a rabbit cyanide toxicity model reveals differences in brain and muscle metabolism

Noninvasive near infrared spectroscopy measurements were performed to monitor cyanide (CN) poisoning and recovery in the brain region and in foreleg muscle simultaneously, and the effects of a novel CN antidote, sulfanegen sodium, on tissue hemoglobin oxygenation changes were compared using a sub-lethal rabbit model. The results demonstrated that the brain region is more susceptible to CN poisoning and slower in endogenous CN detoxification following exposure than peripheral muscles. However, sulfanegen sodium rapidly reversed CN toxicity, with brain region effects reversing more quickly than muscle. In vivo monitoring of multiple organs may provide important clinical information regarding the extent of CN toxicity and subsequent recovery, and facilitate antidote drug development.

Transcript Variants, Expression, and Polymorphisms of the Pig Prosaposin Gene

Prosaposin (PASP) is a sphingolipid hydrolysis protein that plays roles in both the nervous and reproduction systems. In this study, we cloned the pig PASP gene and studied its genomic organization, polymorphism, and expression pattern. Two PASP transcripts, TV1 (HQ245644) and TV2 (HQ245646), were identified in pig. TV1 was the complete transcript that encoded 527 amino acids, whereas TV2 was 9 bp shorter due to an exon 8 deletion. The pig PASP gene spanned over 34 kb in length on chromosome 14 (SSC14), and consisted of 15 exons and 14 introns. The pig PASP gene (TV1 and TV2) expressed predominantly in the cerebellum, lymphnode, pituitary, abdominal fat, hypothalamus, and cerebrum in both females and males. PASP TV1 expressed mainly in teh cerebrum, cerebellum, hypothalamus, pituitary, heart, subcutaneous fat, and foreleg muscle, while TV2 was expressed in the liver, spleen, lung, kidney, and lymphnode. In foreleg muscle, the predominant transcript was TV2 in males and TV1 in females. Some potential transcriptional elements were predicted in 5' flanking region (~3000 bp) of the PASP gene, and they were TATA boxes, RORE, Sp1, SRY, oct-1, Cdx A, and cap. Additionally, we identified 68 single-nucleotide polymorphisms and 9 indels in the pig PASP gene, and three single-nucleotide polymorphisms (C77932320T or L15F; C77928094T or P191L; A77917401G or K522R) were nonsynonymous substitutions. These results provide useful information for future functional investigations of the pig PASP gene.

Detection and surveillance for animal trichinellosis in GB

The zoonotic disease trichinellosis is considered one of the re-emerging diseases with surveillance and control methods constantly gaining more importance worldwide. Recent change in European Union (EU) legislation introduces Trichinella-free production, and the possibility of risk-based monitoring for Trichinella in pigs. This has increased the role of wildlife surveillance programmes and their impact on protecting human health as well as highlighted the need for harmonised surveillance protocols and test methods for these infections. A modified digest method, based on the EU reference method for Trichinella testing of pig meat, was used to screen foxes present in Great Britain (England, Scotland and Wales) for trichinellosis. The method was validated using batched pools of 10 g foreleg muscle from up to 20 foxes (maximum amount 200 g). The method gave an average trichinae recovery rate of 71% for spiked samples. Assuming this recovery rate applies to all contaminated samples, then the test sensitivity would be 70% for all tissue samples with 0.1 trichinae per 10 g of foreleg muscle, 99.9% for samples with 1 trichinae per 10 g, and 100% for samples with 2 or more trichinae per 10 g. In two separate studies, conducted between 1999 to 2001 ( Smith et al., 2003) and 2003 to 2007, over 3500 wild foxes have been screened for Trichinella with negative results. In the second study reported here, foxes were collected from locations throughout Great Britain using a stratified sampling method based on fox population densities. All work was conducted in compliance with appropriate quality assurance systems, latterly under ISO 9001. Results to date indicate the national prevalence of trichinellosis in foxes is <0.001 based on a 10 g individual sample size, an infection level of 1 larva per gram (lpg) and 95% confidence interval. This, together with no reports of trichinellosis in domesticated pigs, suggests that Britain can be considered a region of negligible risk of trichinellosis.

INABILITY TO EASILY PREDICT INDIVIDUAL DEATHS IN A HEMORRHAGE AND RESUSCITATION MODEL

While working on a hemorrhage with low-pressure stabilization protocol, we examined the possibility of identifying individual animals' short term outcomes prior to occurrence using the hemorrhage patterns of several variables currently used or suggested to identify patients and animals as in severe hemorrhagic shock. Methods: 8 anesthetized, ventilated dogs were hemorrhaged (H, 90min mean arterial pressure [MAP] = 35- 40mmHg or MAP < 30mmHg>10min or MAP < 25mmHg>1min), hypotensively stabilized (S, 120min enalaprilat 0.01mg/kg/hr + hemoglobin based oxygen carrier [HBOC] or 7.8% hypertonic saline dextran 70 [HSD] for MAP = 40-45mmHg), resuscitated (60min lactated Ringer's for MAP = 75-80mmHg), then only monitored (60min no fluids), and euthanized. Results: There were 2 non-survivors: NS1 (1 of 5 HBOC, died 117min into S) and NS2 (1 of 3 HSD, died 9min into S). Patterns of continuous, non-invasive variables: Non-survivors had the highest (NS2) and 2nd lowest (NS1) heart rates. All the animals' mean arterial pressures and end-tidal CO2s (related to cardiac output) were completely intermingled. Foreleg muscle StO2 was highest in NS1 (80%s throughout H) and intermingled with the 4 lowest survivors in NS2 (1% for the 1st third of H rising to 10% to 30%s for the remainder of H). Patterns of discontinuous and/or invasive variables: Shed blood volumes in NS1, NS2, and 2 survivors were essentially equivalent (38-40mL/kg) and less than those of the 4 other survivors (45-56mL/kg). All SvO2 values were intermingled. The start H base excess values were intermingled. The end H base excesses were NS1 = −18.8, NS2 = −19.2, 2 survivors each = −19.0, other survivors −13.9 to −17.5 mmol/L. Conclusions: Within our severely hemorrhaged group, the hemorrhage values and obvious patterns of the monitored variables failed to distinguish those individual animals that would undergo cardiovascular collapse despite receiving the same treatment from those that would survive. (Funding: IA Space Grant, Eagles).

Influence of one bout of vigorous exercise on ascorbic acid in plasma and oxidative damage to DNA in blood cells and muscle in untrained rats

We investigated the influence of a single exhaustive bout of downhill running on oxidative damage to DNA and changes of antioxidant vitamin concentrations in rats. Plasma vitamin E levels were unchanged up to 48 hr postexercise. However, plasma ascorbic acid (AA) levels increased after the exercise, then decreased thereafter. This increase corresponded to a marked decrease in AA concentration in the adrenal glands. The activity of hepatic l-gulono-γ-lactone oxidase, which catalyzes AA synthesis, was unaltered after the exercise. The weight of the adrenal glands was significantly increased 24 hr postexercise. These results indicate that the change in the plasma AA concentration after vigorous exercise was due mainly to the release of AA from the adrenal glands. The plasma creatine phosphokinase (CPK) activity and white blood cell (WBC) count increased 3 to 6 hr postexercise. Over this same period, a marker of oxidative DNA damage, 8-hydroxydeoxyguanosine in DNA, increased in the WBC, but not in the foreleg muscle. Lipid peroxide and vitamin E levels were also unchanged in the foreleg muscle. There was a positive correlation between CPK activity in the plasma and DNA damage in the WBC, suggesting that the DNA damage in the WBC was closely related with muscle damage due to exercise.

A Method of Determining the Thermophysical Properties and Calorific Intensity of the Organ or Tissue of a Living Body

A new method was developed to determine simultaneously the thermal conductivity, thermal diffusivity, specific heat, and calorific intensity of the organ or tissue of a living body either in vivo or in vitro with a thin hot probe. By using the method, the thermophysical properties and calorific intensities of a human palm and in vivo liver and a kidney, heart, brain, and foreleg and hindleg muscles of an anesthetized canine were measured. It is concluded that there are no significant differences in the thermophysical properties of organ or tissue of a living body either in vivo or in vitro. The measured thermophysical properties are in good agreement with those reported in the literature.

Carcass characteristics and meat quality of rabbit lines selected for different objectives:: II. Relationships between meat characteristics

Rabbits from three lines (A, V and R) and three different weights (1800, 2050, 2300 g) were used in the experiment. Nine animals of each line and weight were used. Lines A and V were selected for litter size at weaning, whereas line R was selected for growth rate between the 4th and 9th week of life. Muscular pH of Longissimus (pHLD) at the level of the 5th lumbar vertebra and pH of Biceps femoris (pHBF) were taken 24 h post-mortem. Colour (L* lightness, C* chroma, H* hue) was measured on the carcass surface of the Longissimus muscle at the level of the 4th lumbar vertebra and at the 7th lumbar vertebra cut. Fat and moisture were estimated on fore leg muscles (including insertion and thoracic muscles), hind leg, abdominal wall and Longissimus muscle. Water holding capacity, cooking loss (CL) and texture (shear force) were estimated on Longissimus muscle. A principal component analysis was performed in order to examine the relationships between the traits measured. The four 1st PC explained a 63% of the total variation (29%, 16%, 10% and 8%, respectively). The principal component analysis showed that meat quality traits were grouped in independent sets. Fat-moisture, texture, pH and luminosity of Longissimus explained a large part of the observed variation. The differences between lines or liveweight groups in meat quality tend to be small, which implies a certain constancy in rabbit meat quality.

Resistance training-induced increases in muscle mass and performance in ponies.

The purpose of this study was to determine whether 8 wk of progressive resistance exercise training would produce increases in strength and changes in foreleg muscle characteristics indicative of hypertrophy in ponies. Two mature 3- to 6-yr-old, male ponies (188 +/- 16 kg) were taught to carry sheets of lead over their saddle region (wither) while walking on a level treadmill at 1.9 m.s-1. This initial familiarization period was followed by 8 wk of training (3 d per wk), in which the ponies performed a series of progressive sets of weight carrying to fatigue. Each workout started with a 2-min walk at 1.9 m.s-1 followed by sets of weight carrying. The ponies carried 44.5 kg for the first set with increases of 22.3 kg per set until fatigue. Weights were applied and then removed for 60-90 s between sets using a chain hoist and sling apparatus. Measurements of forelimb girth, body weight, and total weight carried were recorded at each workout session. Ultrasound measurement of the diameters of the superdigital flexor muscles and muscle biopsies were performed before and after the 8-wk training period. Eight weeks of resistance training resulted in significant increases in peak weight carried (260%, P < 0.05) and total weight carried (1525%, P < 0.05) during each workout. Forelimb girth increased 12 +/- 1% (P < 0.05) with a corresponding 19 +/- 3% (P < 0.05) increase in muscle cross-sectional diameter. There were no changes (P > 0.05) in Type I muscle fiber area; however, there was a nonsignificant 26% increase in Type IIA+IIB fiber area. These data suggest that 8 wk of progressive resistance exercise training increase strength and cause changes in muscle size and characteristics consistent with hypertrophy.

The relationship between the degree of paw preference and excitability of motor neurons innervating foreleg flexors in right- and left-preferent cats

The relationship between the degree of paw preference and the degree of excitability of motor neurons innervating the foreleg flexor muscles was studied in unanesthetized spinal cats. The paw preference was assessed by a food-reaching test. The cats were spinalized at the first cervical segment of the spinal cord. It was found that there is an inverse relationship between the degree of H-reflex excitability and the degree of paw preference in right- and left-preferent cats. This main result was discussed with respect to functional implications and evolution of human handedness.

Acid protease activity in spinal cord and muscle in wobbler mouse

Acid protease activity is increased in spinal cord and foreleg muscles in wobbler mice with motor neuron disease as compared to control mice from the wobbler colony. The control mice from the wobbler colony, which contain a large proportion of heterozygotes have higher level of acid protease activity in spinal cord, but not muscle, than mice from other strains.

Some evidence for concurrent involvement of the fore- and hindleg muscles in murine muscular dystrophy.

Endurance of the forelegs of the dystrophic mouse was found to be significantly less than that of the normal mouse as early as at 3 weeks of age. It decreased progressively with age although the forelegs seemed to function almost normally even after 2 months of age by which time the hindlegs became almost immobile. Thus, histological and biochemical studies to compare the fore- and hindleg muscles of the dystrophic mice were conducted. Morphological abnormalities similar to those observed in the hindleg muscle were also found in foreleg muscles of the young dystrophic mice. Their foreleg and hindleg muscles showed significantly higher hydroxyproline contents than those of the normal mice even at 2 weeks of age when clinical signs of the disease first became manifest in their hindlegs. The present findings appear to be the first evidence for concurrent involvement of the fore- and hindleg muscles in murine muscular dystrophy. This would raise an issue as to why forelegs with abnormal muscles can subserve somewhat normal function, whereas hindlegs with abnormal muscles can subserve somewhat function, whereas hindlegs in these adult dystrophic mice are immobile. This might yield clues to clarify the mechanism underlying the aggravation of the disease.

Relations between the effect of tetanus toxin on the neuromuscular transmission and histological functional properties of various muscles of the rat.

Various doses of tetanus toxin were injected into three hind leg and two fore leg muscles of the rat. The neuromuscular transmission was tested by recording the mass action potential of the muscles elicited by a single electrical stimulus to the motor nerve after strong symptoms of local tetanus had developed. The muscle responses were depressed and blocked at lower toxin doses in the fast tibialis anterior than in the mixed gastrocnemius latemlis, while blocking of the slow soleus required the highest dose. The extensor carpi radialis and the flexor carpi ulnaris muscles showed medium sensitivity. In all five muscles the contraction time was measured and correlated with its individual minimal blocking dose. The more phasic (i.e., the faster) the muscle, the more sensitive its neuromuscular transmission was to tetanus toxin. The proportional distribution of red, white, and intermediate fibres, which are associated with specific end-plate types, was evaluated for the five muscles. The percentage of white fibres in the muscles displayed a very good negative correlation with the blocking dose. The relation between structures of end-plates and effects of tetanus toxin were analysed and it is suggested that the differences in sensitivity to tetanus toxin in the neuromuscular transmission in the five muscles is determined by a differential distribution of endplates with varying sensitivities to this toxin due to structural properties.