Comparative proteomic study of pig muscle proteins during growth and development of an animal

The production of high-quality pork is closely related to the growth and development of muscle tissue. The present article provides a comparative proteomic research of l. dorsi, b. femoris, m. brachiocephalicus during the pigs’ growth and development (at age of 60 days and 180 days). This work was supported by data of electrophoretic methods: one-dimensional electrophoresis according to Laemmli with densitometric assessment in the ImageJ software and two-dimensional electrophoresis according to O’Farrell method with its further processing on the software ImageMaster. The mass spectrometric identification was conducted with the help of the high-performance liquid chromatography (HPLC) system connected to a mass spectrometer; further the data were interpreted by search algorithm Andromeda. When comparing frequency diagrams of one-dimensional electrophoregrams of all three muscle tissues of weaned pigs, the greatest difference was observed for the muscle sample l. dorsi. Comparison of diagrams of muscle tissue samples taken for mature pigs showed a great similarity of all three studied muscles samples. Within the framework of the research, the Fold indicator was calculated. The exceeding its value by more than 2 units is generally considered to be a statistically significant difference. When analyzing two-dimensional electrophoretograms of weaned pigs’ muscles, 18 protein fractions were revealed with Fold > 2. When examining the muscle tissue of mature pigs, 15 of those proteins were found; the differences were mostly detected in the minor protein fractions. The mass spectrometric analysis of the cut bands with well-pronounced differences from the onedimensional electrophoretogram revealed 214 proteins involved to a greater extent in cellular and metabolic processes, physical activity and localization. Growth and development protein — semaphorin‑6B (96.78 kDa) — was revealed in muscle tissue of l. dorsi, a. Also in l. dorsi and b. femoris the growth and development proteins were found: cadherin‑13 (78.23 kDa), cadherin‑7 (87.01 kDa), the F‑actin-cap protein beta subunit (30.66 kDa), and two uncharacterized proteins at 65.60 kDa and 63.88 kDa.

Quality and Safety of Game Meat from the Biocenosis of the Beloosipovo Mercury Deposit

Introduction. Mercury contamination is one of the most common environmental problems. The research objective was to study the qualitative composition and physicochemical properties of raw game meat obtained from the area near the Beloosipovo mercury deposit in order to define any possible contamination w ith xenobiotics. Study objects and methods. The research featured rib eye muscle tissue and soft flesh of elks shot on the hunting farms of the Kemerovo Region aka Kuzbass. Results and discussion. A complex set of experiments revealed the chemical composition of elk muscle tissue and flesh, as well as the mineral composition of elk muscle tissue. The samples were obtained from different parts of carcasses. The amino acid and fatty acid composition of elk muscle tissue made it possible to describe the biological value, mineral composition, and vitamin profile of elk meat. The physicochemical analysis included toughness, cooking losses, and moisture-retaining capacity, i.e. the properties that ensure juiciness. The research also featured the accumulation of xenobiotics in elk meat samples obtained from the biosinosis near the Beloosipovo merc ury deposit. Conclusion. The slaughter yield of elk meat was 51–53%, which exceeds the average yield of farm cattle meat by 4–6%. The moisture content was 73–78%, while the content of protein was between 20–24% and depended on the anatomical location of the muscle sample; the fat content reached 0.75–1.75%. The mercury accumulation at different storage temperature conditions ranged from 0.004 ± 0.001 to 0.009 ± 0.001 mg/kg, while the max imum allowable concentration of mercury is 0.03 mg/kg.

Biochemical Profile and Antibacterial Examination of Freshwater Crab Scylla Serrata (FORSKAL, 1775)

Natural products from the aquatic organisms provide unlimited opportunities for the production of new drugs. To verify this fact in the present study ‘the biochemical profile and antimicrobial activity of different samples of the estuarine crab, Sylla serrata’ was investigated. Biochemical study of the body parts of the crab gains substantial attention for their specificity in connection to the food values of the crab and for the evaluation of their physiological needs at different periods of life. The total protein was estimated by Lowry's method and the amount of protein was highest in the soft muscle sample (20.27mg/ml). The total sugar was estimated by the Phenol-Sulphuric acid method and it was highest in the leg sample (209.8mg/ml). The lipids of the samples were estimated by Folchs method. Lipid was highest in the shell sample (28.35mg/ml). Thin Layer chromatography was done for characterization of protein. Antibiotics are used against bacterial infections. The natural products gained importance to screen antibiotics. For this reason the antibacterial activity of the samples were tested against grampositive and gramnegative pathogenic bacterial strains. The bacterial strains like Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Enterobacter aerogenes were used as the test strains. In the present study, the ethanol extract of crab sample I showed maximum zone of inhibition (13 mm) in P. aeruginosa. The sample I showed moderate zone of inhibition (11 mm) in K.pneumoniae. The sample II showed the minimum zone of inhibition (7 mm) in P. aeruginosa. It indicated that the shell of crabs would be a good source of antimicrobial agents and would replace the existing inadequate and cost effective antibiotics. The microbial associates of crustaceans have proven to be a rich source of bioactivity with antimicrobial and cytotoxic effects. So the organism used in the present study will be the promising source to the biotechnological and pharmaceutical applications.

Enhancing Interrogation of Skeletal Muscle Samples for Informative Quantitative Data

Careful quantitative analysis of histological preparations of muscle samples is crucial to accurate investigation of myopathies in man and of interpretation of data from animals subjected to experimental or potentially therapeutic treatments. Protocols for measuring cell numbers are subject to problems arising from biases associated with preparative and analytical techniques. Prominent among these is the effect of polarized structure of skeletal muscle on sampling bias. It is also common in this tissue to collect data as ratios to convenient reference dominators, the fundamental bases of which are ill-defined, or unrecognized or not accurately assessable. Use of such ‘floating’ denominators raises a barrier to estimation of the absolute values that assume practical importance in medical research, where accurate comparison between different scenarios in different species is essential to the aim of translating preclinical research findings in animal models to clinical utility in Homo sapiens. This review identifies some of the underappreciated problems with current morphometric practice, some of which are exacerbated in skeletal muscle, and evaluates the extent of their intrusiveness into the of building an objective, accurate, picture of the structure of the muscle sample. It also contains recommendations for eliminating or at least minimizing these problems. Principal among these, would be the use of stereological procedures to avoid the substantial counting biases arising from inter-procedure differences in object size and section thickness. Attention is also drawn to the distortions of interpretation arising from use of undefined or inappropriate denominators.

IMPACT OF GREEN FORAGES IN SPRING, IN THE STEPPE REGIONS OF TIARET “CHEHAIMA AND NAIMA”, ON THE FATTY ACID COMPOSITION OF THE "BICEPS FEMORIS" MUSCLE OF LAMBS

This work aimed to assess the impact of a diet based on green pastures of steppe rangelands and another based on a concentrate diet, on the composition of the intramuscular lipid fraction of different sections of the biceps femoral muscle of lambs, in two regions of the wilaya of Tiaret, in winter and in spring. The biceps femoral muscle sample collection came from 20 different lamb carcasses (10 samples for each of our two study regions, with 05 samples for each season). These samples were taken on different occasions in large butcher shops. Total SFA content in both regions shows dominance in lambs from the regimen concentrate, that C 16: 0 and C 18: 0 are the most dominant. There is no remarkable difference in total MUFA content, and on individual monounsaturated FA, between the two regimens. In both regions, the most dominant FAs in the intramuscular lipid fraction of pasture lambs are C18: 3n3 followed by C18: 2n6 cis-9, C20: 4n6, C20: 5n3, and C22: 6n3, compared to those from concentrate regimen; however, lambs from Chehaima concentrate regimen had the most dominant FA in their intramuscular fat, C18: 3n6 followed by C20: 4n6, compared to those from pastures of Naima. The values of the ratio n-6 / n-3 correspond to the nutritional recommendations. The feed ratio ΣPUFA / ΣSFA of lambs from pastures in the Chehaima region is the only one that corresponds to the desired value. Based on this research, it is concluded that lambs that have grazed grass in steppe rangelands have the richest meat in omega 3, which is more desirable and benefits for human health.

Organochlorine Pesticides Occurrence In Fish Muscle And Their Public Health Risks In Northwest Ethiopia

Pesticides are the parent compounds, their metabolites, and associated impurities of agricultural and health chemical inputs. If they are found concentration level higher than the standard limits, they have potential negative impacts on ecosystem in general and in fish and humans in particular. This study investigates organochlorine pesticides (OCPs) residue occurrences in fish muscle and assesses their public health potential risks, in North West Ethiopia. The concentration of OCPs residue under GC-ECD detected in 37.84% fish muscle samples. The mean amount detected were Endosalfan I, 341.50 ± 32.19 μg/kg; Endosalfan II, 36.01 ± 2.3μg/kg; Endosalfan sulfate, 5.43 ± 4.06 μg/kg; 4,4, DDE, 64.01 ± 9.08μg /kg; 4, 4, DDD, 5.65 ± 3.12μg /kg; and 4,4, DDT, 1.58 ± 0.30μg/kg. The mean concentration of Endosalfan I tested fish muscle sample was higher than that of permissible limit of different international standards. However, due to low per capital consumption rate of fish origin food in Ethiopia, health risk index (HRI) ranges from 0.002-0.1275 which shows there is no public health risk. This study highlights the possibilities of chemical residues occurrence in fish food products and hence pesticide use regulations and monitoring concentration level should be implemented regularly to avoid human and environmental health risks.

Application of shearwave™ elastography to evaluate heat-induced changes in the young's modulus of fresh bovine muscle: A preliminary study

PurposeThe purpose of this study was to investigate the variation in the Young’s modulus (E) of bovine muscle samples as a function of temperature change generated by therapeutic ultrasound using Shearwave™ Elastography.MethodsInitially, the bovine muscle was heated via therapeutic ultrasound with a frequency of 3 MHz, nominal intensity of 2 W·cm-2, and application time of 2 min. Immediately following cessation of therapeutic irradiation, an E image was recorded and the stiffness was measured in circular area positioned at six depths (from 0.4 to 2.9 cm) in the center of the region of interest. Next, an E image was recorded every minute for the first 5 min. Over the next 30 min, an image was recorded every 5 min. Finally, an image was acquired 60 min after cessation of therapeutic irradiation. In the second test, the same experimental procedure was performed 60 min later with the physiotherapy equipment configured with a 10-min application time. Finally, during the ultrasonic irradiation of a new bovine muscle sample, the physiotherapeutic transducer was applied in a circular motion and with an angular velocity of 3.6 ± 0.3 rad·s-1.ResultsIn the first test, the bovine muscle E decreased from 212.2 ± 32.8 to 80.1 ± 13.8 kPa at 0.4 cm-depth, as the temperature increased from 18.2 to 44.9 °C. This effect was reversed when the temperature decreased. In the second test, denaturation and cell death occurred, so an artifact appeared in the elastographic image and the Shearwave™ Elastography did not capture the E from the depth of 1.9 cm.ConclusionWe confirmed that it is possible to use Shearwave™ Elastography to investigate heat-induced changes in the elastic modulus of biological tissue.

Myoglobin Concentration and Oxygen Stores in Different Functional Muscle Groups from Three Small Cetacean Species

Compared with terrestrial mammals, marine mammals possess increased muscle myoglobin concentrations (Mb concentration, g Mb · 100g−1 muscle), enhancing their onboard oxygen (O2) stores and their aerobic dive limit. Although myoglobin is not homogeneously distributed, cetacean muscle O2 stores have been often determined by measuring Mb concentration from a single muscle sample (longissimus dorsi) and multiplying that value by the animal’s locomotor muscle or total muscle mass. This study serves to determine the accuracy of previous cetacean muscle O2 stores calculations. For that, body muscles from three delphinid species: Delphinus delphis, Stenella coeruleoalba, and Stenella frontalis, were dissected and weighed. Mb concentration was calculated from six muscles/muscle groups (epaxial, hypaxial and rectus abdominis; mastohumeralis; sternohyoideus; and dorsal scalenus), each representative of different functional groups (locomotion powering swimming, pectoral fin movement, feeding and respiration, respectively). Results demonstrated that the Mb concentration was heterogeneously distributed, being significantly higher in locomotor muscles. Locomotor muscles were the major contributors to total muscle O2 stores (mean 92.8%) due to their high Mb concentration and large muscle masses. Compared to this method, previous studies assuming homogenous Mb concentration distribution likely underestimated total muscle O2 stores by 10% when only considering locomotor muscles and overestimated them by 13% when total muscle mass was considered.

Expression of titin-linked putative mechanosensing proteins in skeletal muscle after power resistance exercise in resistance-trained men

Little is known about the molecular responses to power resistance exercise that lead to skeletal muscle remodeling and enhanced athletic performance. We assessed the expression of titin-linked putative mechanosensing proteins implicated in muscle remodeling: muscle ankyrin repeat proteins (Ankrd1, Ankrd2 and Ankrd23), muscle-LIM proteins (MLP), muscle RING-finger protein-1 (MuRF-1), and associated myogenic proteins (MyoD1, myogenin, and myostatin) in skeletal muscle in response to power resistance exercise with or without a post-exercise meal, in fed, resistance-trained men. A muscle sample was obtained from the vastus lateralis of seven healthy men on separate days, 3h after 90 min of rest (Rest) or power resistance exercise with (Ex+Meal) or without (Ex) a post-exercise meal, to quantify mRNA and protein levels. The levels of phosphorylated HSP27 (pHSP27-Ser15) and cytoskeletal proteins in muscle and creatine kinase activity in serum were also assessed. The exercise increased (P≤0.05) pHSP27-Ser15 (~6-fold) and creatine kinase (~50%), whereas cytoskeletal protein levels were unchanged (P>0.05). Ankrd1 (~15-fold) and MLP (~2-fold) mRNA increased, whereas Ankrd2, Ankrd23, MuRF-1, MyoD1, and myostatin mRNA were unchanged. Ankrd1 (~3-fold, Ex) and MLPb (~20-fold, Ex+Meal) protein increased, but MLPa, Ankrd2, Ankrd23, and the myogenic proteins were unchanged. The post-exercise meal did not affect the responses observed. Power resistance exercise, as performed in practice, induced subtle early responses in the expression of MLP and Ankrd1, yet had little effect on the other proteins investigated. These findings suggest possible roles for MLP and Ankrd1 in the remodeling of skeletal muscle in individuals who regularly perform this type of exercise.