Physicochemical, thermal and mechanical characterization study of perimysial collagen of two bovine muscles.

Chemical, thermal and mechanical collagen characteristics of intramuscular perimysial connective tissue (IMCT) from bovine Semitendinosus (ST) and Pectoralis profundus (PP) muscles were studied. Furthermore, these collagen characteristics in presence/absence of other extracellular matrix components were analyzed for both muscles. Differences between muscles were observed for collagen content; all IMCT-PP perimysial samples were higher than ST samples. In addition, for both muscles, IMCT-alkali resistant samples allowed the highest trypsin soluble collagen. The main differences between muscles were recorder for thermal and mechanical properties. The denaturation of collagen in the perimysium evidenced differences in total denaturation energy (ΔH) and peak temperatures (Tp). The ΔH resulted higher for IMCT-PP than for IMCT-ST tissues in all samples. By the tensile test it was observed that the maximum loads were constant and higher in all PP samples. In the FTIR assay, the peaks for the main amides were registered in both tissues. However, slight differences between ST and PP-IMCT were detected on hydrogen bond interactions and in secondary structure of the protein. The results reinforce the hypothesis of the presence of different IMCT-perimysial-collagen pools. In this study, chemical, thermal and mechanical characteristics were considered and quantified. However, the mechanical function and development of muscle in-vivo could be the main influence on the extracellular collagen characteristics as well as its interactions with non-collagen compounds. Its formation is essential for muscle function.

Specific effects on strength and heat stability of intramuscular connective tissue during long time low temperature cooking.

Long-time low-temperature (LTLT) cooking of meat is known to produce a tender product. The current work tested the hypothesis that LTLT cooking for periods of up to 24 h at 60 °C reduces the contribution of intramuscular connective tissue to cooked meat toughness. Tensile tests on perimysium excised after cooking showed that its strength diminished with cooking time, although not as markedly as the Warner-Bratzler peak force measure of toughness. A gradually increasing susceptibility to trypsin digestion with increasing heating time demonstrated that there was a slow and gradual increase in the proportion of denatured collagen in the perimysium. Differential scanning calorimetry on perimysium excised after cooking showed an endothermic peak representing the denaturation of the collagen not already denatured on cooking. With increasing cooking time, the energy per milligram of collagen necessary to denature this remaining fraction increased. These results support the hypothesis that there is both an easily destabilized and more resistant fractions of the collagen in intramuscular connective tissue.

Differences in the energetics of collagen denaturation in connective tissue from two muscles.

The thermal denaturation of collagen the perimysium of intramuscular connective tissue isolated from bovine Semitendinosus (ST) and Perctoralis profundus (PP) muscles was investigated using a range of heating rates in differential scanning calorimetry (DSC) and analyzed by application of the Kissinger-Akahira-Sunose (KAS) and Lumry-Eyring models. Thermograms showed a broadening of endotherms and a shift towards higher temperatures as the thermal scanning rate increased. These features are consistent with the two-step process of a reversible transition between native and unfolded collagen molecules followed by an irreversible transition between unfolded and denatured states. There were small differences between muscles in the onset temperatures of the thermal transitions at heating rates of 0.5 K min-1, while both the KAS and Lumry-Eyring models yielded similar values for the effective activation energy of the whole two-step process, the Lumry-Eyring model allowed a greater insight into differences in the reversible and irreversible steps between the perimysium both muscles. Reversible unwinding of the triple-helical collagen molecules in the perimysium from ST muscle required more energy than in PP muscle. It is speculated that the presence of large amounts of elastin in the perimysium of ST muscles may influence this due to a protein crowding mechanism, or by affecting the covalent cross-linking of the collagen.

The thermal shrinkage force in perimysium from different beef muscles is not affected by post-mortem ageing.

Differences in the thermal shrinkage and collagen solubility between bovine Semitendinosus (ST) and Pectoralis profundus (PP) muscles and their interactions with ageing were evaluated by studying collagen solubility, hydrothermal isometric tension and thermal denaturation properties of intramuscular connective tissue after 5-20days post-mortem storage at 4°C. Collagen solubility was higher in ST than in PP muscle at 5-13days, but the differences between the two muscles decreased at longer ageing times. A small decrease in the peak denaturation temperature of perimysium occurred with increasing ageing times in both muscles. Maximum force in isometrically-heated perimysium was broadly equivalent in both muscles. Although the amount and solubility of collagen varies between muscles and ageing decreases the stability of some of the collagen, thermal shrinkage forces in heated perimysium are not significantly diminished by ageing. These findings support the idea of one collagen fraction easily degraded by ageing and heat, and another more resistant fraction that determines the physical properties of the tissue after ageing and cooking.

New recommendations for measuring collagen solubility.

The heat-solubility of intramuscular collagen is usually conducted in 1/4 Ringer's solution at pH7.4, despite this ionic strength and pH being inappropriate for post-rigor meat. The current work studied the percentage of soluble collagen and hydrothermal isometric tension characteristics of perimysial strips on bovine semitendinosus muscles in either 1/4 Ringer's solution, distilled water, PBS, or a solution of the same salt concentration as 1/4 Ringer's but at pH5.6. Values of % soluble collagen were lower at pH7.4 than 5.6. Increasing ionic strength reduced % soluble collagen. The maximum perimysial isometric tension was independent of the bathing medium, but the percent relaxation was higher at pH7.4 than at pH5.6, and increased with ionic strength of the media. It is recommended that future measurements of collagen solubility and tests on connective tissue components of post-rigor meat should be carried out in a solution of concentrations NaCl and KCl equivalent to those in 1/4 Ringer's, but at pH5.6, a pH relevant to post-rigor meat.

Conjugated linoleic acid improves glucose utilization in the soleus muscle of rats fed linoleic acid-enriched and linoleic acid-deprived diets.

The effect that conjugated linoleic acid (CLA) has on glucose metabolism in experimental animals depends on nutritional conditions. Therefore, we hypothesized that CLA improves glucose utilization and insulin sensitivity in rats fed different levels of dietary linoleic acid (LA). We investigated the effect of CLA on the uptake, incorporation, and oxidation of glucose and glycogen synthesis in the soleus muscle of rats who were fed either LA-enriched (+LA) or LA-deprived (-LA) diets, under basal conditions and in the absence or presence of insulin and/or palmitate. For 60 days, male Wistar rats were fed 1 of 4 diets consisting of +LA, -LA, or +LA and -LA supplemented with CLA. Nutritional parameters and soleus glucose metabolism were evaluated. Under basal conditions, CLA enhanced soleus glucose oxidation, whereas increased glucose uptake and incorporation were observed in the -LA + CLA group. Conjugated linoleic acid-supplemented rats presented a lower response to insulin on glucose metabolism compared with non-CLA-supplemented rats. Palmitate partially inhibited the effect of insulin on the uptake and incorporation of glucose in the +LA and -LA groups but not in the +LA + CLA or -LA + CLA groups. Dietary CLA increased glucose utilization under basal conditions and prevented the palmitate-induced inhibition of glucose uptake and incorporation that is stimulated by insulin. The beneficial effects of CLA were better in LA-deprived rats. Conjugated linoleic acid may also have negative effects, such as lowering the insulin response capacity. These results demonstrate the complexities of the interactions between CLA, palmitate, and/or insulin to differentially modify muscle glucose utilization and show that the magnitude of the response is related to the dietary LA levels.

Effect of gamma irradiation and storage time on microbial growth and physicochemical characteristics of pumpkin (Cucurbita Moschata Duchesne ex Poiret) puree.

The effect of gamma irradiation (0-2 kGy) and storage time (0-28 days) on microbial growth and physicochemical characteristics of a packed pumpkin puree was studied. For that purpose, a factorial design was applied. The puree contained potassium sorbate, glucose and vanillin was stored at 25°C . Gamma irradiation diminished and storage time increased microbial growth. A synergistic effect between both variables on microbial growth was observed. Storage time decreased pH and color of purees. Sorbate content decreased with storage time and gamma irradiation. Mathematical models of microbial growth generated by the factorial design allowed estimating that a puree absorbing 1.63 kGy would have a shelf-life of 4 days. In order to improve this time, some changes in the applied hurdles were assayed. These included a thermal treatment before irradiation, a reduction of irradiation dose to 0.75 kGy and a decrease in storage temperature at 20°C . As a result, the shelf-life of purees increased to 28 days.