Inhibition of pyruvate dehydrogenase accelerates anaerobic glycolysis under postmortem simulating conditions.

This research aimed to explore the potential influence of mitochondria on the rate of anaerobic glycolysis. We hypothesized that mitochondria could reduce the rate of anaerobic glycolysis and pH decline by metabolizing a portion of glycolytic pyruvate. We utilized an in vitro model and incorporated CPI-613 and Avidin to inhibit pyruvate dehydrogenase (PDH) and pyruvate carboxylase (PC), respectively. Four treatments were tested: 400 µM CPI-613, 1.5 U/ml Avidin, 400 µM CPI-613 + 1.5 U/ml Avidin, or control. Glycolytic metabolites and pH of the in vitro model were evaluated throughout a 1440-min incubation period. CPI-613-containing treatments, with or without Avidin, decreased pH levels and increased glycogen degradation and lactate accumulation compared to the control and Avidin treatments (P < 0.05), indicating increased glycolytic flux. In a different experiment, two treatments, 400 µM CPI-613 or control, were employed to track the fates of pyruvate using [13C6]glucose. CPI-613 reduced the contribution of glucose carbon to tricarboxylic acid cycle intermediates compared to control (P < 0.05). To test whether the acceleration of acidification in reactions containing CPI-613 was due to an increase in the activity of key enzymes of glycogenolysis and glycolysis, we evaluated the activities of glycogen phosphorylase, phosphofructokinase, and pyruvate kinase in the presence or absence of 400 µM CPI-613. The CPI-613 treatment did not elicit an alteration in the activity of these three enzymes. These findings indicate that inhibiting PDH increases the rate of anaerobic glycolysis and pH decline, suggesting that mitochondria are potential regulators of postmortem metabolism.

Spectro 1-A Potential Spectrophotometer for Measuring Color and Myoglobin Forms in Beef.

The objective of this study was to compare the Color Muse Spectro 1 sensor to the HunterLab MiniScan XE Plus spectrophotometer for evaluating beef color. Color coordinates (lightness (L*), redness (a*), yellowness (b*), chroma (C*), and hue (h*)), myoglobin redox forms (metmyoglobin (MMb), deoxymyoglobin (DMb), and oxymyoglobin (OMb)), and metmyoglobin reducing ability (MRA) were measured on beef steaks over a 5-days storage period. The results indicated that L*, b*, C*, MMb%, OMb%, and MRA% values obtained with Spectro 1 were comparable to those of MiniScan. However, Spectro 1 values for a* were overestimated compared to MiniScan (p < 0.05), whereas those for h* and DMb% were underestimated (p < 0.05). Regardless, Spectro 1 had the capability to detect changes in color coordinates, myoglobin forms, and MRA throughout the storage period. Bland−Altman plots demonstrated that L*, b*, and C* are interchangeable between the two instruments, but it was not the case for a*, h*, myoglobin forms, and MRA. Color coordinates measured by Spectro 1 exhibited excellent stability over time, evidenced by the low total color difference (ΔE*ab) values. Collectively, these findings indicate that Spectro 1 is a potential alternative spectrophotometer for studying meat color and myoglobin redox forms.

Cattle breed type and anabolic implants impact calpastatin expression and abundance of mRNA associated with protein turnover in the longissimus thoracis of feedlot steers.

Two methods that the beef cattle industry can use to improve efficiency, sustainability, and economic viability are growth promotants and crossbreeding cattle of different breed types. In the United States, over 90% of cattle receive an anabolic implant at some point during production resulting in an overall increase in skeletal muscle growth. Recent research suggests that the two main cattle breed types, Bos indicus and Bos taurus, respond differently to anabolic implants. The objective of this study was to characterize changes that occur in skeletal muscle following implanting in Bos indicus influenced steers or Bos taurus steers. Twenty steers were stratified by initial weight in a 2 × 2 factorial design examining two different breeds: Angus (AN; n = 10) or Santa Gertrudis influenced (SG; n = 10), and two implant strategies: no implant (CON; n = 10) or a combined implant containing 120 mg TBA and 24 mg E2 (IMP; n = 10; Revalor-S, Merck Animal Health). Skeletal muscle biopsies were taken from the longissimus thoracis (LT) 2 and 10 d post-implantation. The mRNA abundance of 24 genes associated with skeletal muscle growth were examined, as well as the protein expression of µ-calpain and calpastatin. Succinate dehydrogenase mRNA abundance was impacted (P = 0.05) by a breed × treatment interaction 2 d post-implanting, with SG-CON having a greater increased abundance than all other steers. A tendency for a breed × treatment interaction was observed for calpain-6 mRNA (P = 0.07), with SG-CON having greater abundance than AN-CON and SG-IMP. Additionally, calpastatin protein expression was altered (P = 0.01) by a breed × treatment interaction, with SG-CON and SG-IMP steers having increased expression (P = 0.01) compared with AN-CON steers. At 2 d post-implanting, a breed × treatment interaction was observed with SG-CON steers having greater (P = 0.05) mRNA abundance of mitogen-activated protein kinase compared with AN-CON steers. Furthermore, breed affected (P = 0.05) calpastatin abundance with AN steers having increased (P = 0.05) abundance 2 d post-implanting compared with SG steers. Meanwhile, implants tended to affect (P = 0.09) muscle RING finger protein-1 mRNA abundance, with CON steers having increased (P = 0.09) abundance compared with that of IMP steers. These findings suggest that cattle breed type and anabolic implants impact calpastatin expression and mRNA abundance associated with protein turnover in the LT of feedlot steers 2 and 10 d post-implantation.

Two methods that the beef cattle industry can use to potentially improve efficiency, sustainability, and economic viability are growth promotants and crossbreeding cattle of different breed types. In the United States, over 90% of cattle receive at least one anabolic implant during the production cycle resulting in improvements in production and overall economic and environmental sustainability. Research suggests that the two main cattle breed types, Bos indicus and Bos taurus, respond differently to different anabolic implant strategies. The objective of this study was to characterize changes that occur in the skeletal muscle following implanting in Bos indicus influenced animals and Bos taurus animals. This research measured mRNA abundance of 24 genes associated with skeletal muscle growth, and protein expression of calpain-1 and calpastatin. The findings of this research suggest that anabolic implants and cattle breed type interact to cause changes in mRNA abundance in the longissimus thoracis that are related to protein turnover of skeletal muscle. Furthermore, calpastatin protein abundance was also altered by this breed × treatment interaction. This research demonstrates that anabolic implants cause molecular changes in skeletal muscle of feedlot steers, with some of these changes being breed dependent.

Tandem mass tag labeling to assess proteome differences between intermediate and very tender beef steaks.

Tenderness is considered as one of the most important quality attributes dictating consumers' overall satisfaction and future purchasing decisions of fresh beef. However, the ability to predict and manage tenderness has proven very challenging due to the numerous factors that contribute to variation in end-product tenderness. Proteomic profiling allows for global examination of differentially abundant proteins in the meat and can provide new insight into biological mechanisms related to meat tenderness. Hence, the objective of this study was to examine proteomic profiles of beef longissimus lumborum (LL) steaks varying in tenderness, with the intention to identify potential biomarkers related to tenderness. For this purpose, beef LL muscle samples were collected from 99 carcasses at 0 and 384 h postmortem. Based on Warner-Bratzler shear force values at 384 h, 16 samples with the highest (intermediate tender, IT) and lowest (very tender, VT) values were selected to be used for proteomic analysis in this study (n = 8 per category). Using tandem mass tag-based proteomics, a total of 876 proteins were identified, of which 51 proteins were differentially abundant (P < 0.05) between the tenderness categories and aging periods. The differentially identified proteins encompassed a wide array of biological processes related to muscle contraction, calcium signaling, metabolism, extracellular matrix organization, chaperone, and apoptosis. A greater (P < 0.05) relative abundance of proteins associated with carbohydrate metabolism and apoptosis, and a lower (P < 0.05) relative abundance of proteins involved in muscle contraction was observed in the VT steaks after aging compared with the IT steaks, suggesting that more proteolysis occurred in the VT steaks. This may be explained by the greater (P < 0.05) abundance of chaperonin and calcium-binding proteins in the IT steaks, which could have limited the extent of postmortem proteolysis in these steaks. In addition, a greater (P < 0.05) abundance of connective tissue proteins was also observed in the IT steaks, which likely contributed to the difference in tenderness due to added background toughness. The established proteomic database obtained in this study may provide a reference for future research regarding potential protein biomarkers that are associated with meat tenderness.

Among all the eating quality attributes of beef, tenderness is considered an essential factor influencing consumers' overall satisfaction and future purchasing decisions. However, managing and predicting tenderness of meat products is challenging for the meat industry, as many factors can influence this attribute. The goal of this research was to examine variations in protein abundance between two categories of beef strip steaks varying in tenderness, with the intention to identify proteins related to beef tenderness/toughness. Overall, the results from this study suggest that tender steaks experienced greater protein degradation during aging than tougher steaks, which likely contributed to their improved tenderness. Furthermore, a greater abundance of connective tissue proteins, which are associated with meat toughness, was observed in the tougher steaks. Our results collectively indicate that the difference in tenderness between the two groups of steaks may be due to multiple proteins involved in several biological processes.

Ultrasonication of beef improves calpain-1 autolysis and caspase-3 activity by elevating cytosolic calcium and inducing mitochondrial dysfunction.

The objective of this study was to investigate if ultrasonication of bovine longissimus thoracis et lumborum (LTL) steaks increases calpain-1 and caspase-3 activities, and if so, to explore the underlying mechanisms that trigger their activation. Post-rigor bovine LTL steaks were subjected to ultrasonication at 40 kHz and 12 W/cm2 for 40 min and subsequently aged for 14 d at 4 °C. Ultrasonication improved beef tenderness (P < 0.05) without negatively impacting pH, color, or cook loss (P > 0.05). Improved tenderness in the ultrasonicated steaks was associated with greater degradation of titin, desmin, troponin-T, and calpastatin and increased calpain-1 autolysis and caspase-3 activity (P < 0.05). In addition, ultrasonicated steaks had greater levels of cytosolic calcium and reactive oxygen species and lower mitochondrial oxygen consumption rate (P < 0.05). These data indicate that improved beef tenderness following ultrasonication is, in part, a function of increased calpain-1 and caspase-3 activities, potentially by elevating cytosolic calcium and inducing mitochondrial dysfunction, respectively.

Crystallization Behavior and Quality of Frozen Meat.

Preservation of meat through freezing entails the use of low temperatures to extend a product's shelf-life, mainly by reducing the rate of microbial spoilage and deterioration reactions. Characteristics of meat that are important to be preserve include tenderness, water holding capacity, color, and flavor. In general, freezing improves meat tenderness, but negatively impacts other quality attributes. The extent to which these attributes are affected depends on the ice crystalline size and distribution, which itself is governed by freezing rate and storage temperature and duration. Although novel technology has made it possible to mitigate the negative effects of freezing, the complex nature of muscle tissue makes it difficult to accurately and consistently predict outcome of meat quality following freezing. This review provides an overview of the current understanding of energy and heat transfer during freezing and its effect on meat quality. Furthermore, the review provides an overview of the current novel technologies utilized to improve the freezing process.

Injection of iodoacetic acid into pre-rigor bovine muscle simulates dark cutting conditions.

The purpose of this study was to develop an in situ model for dark cutting beef. Iodoacetic acid (IAA) was injected at different concentrations (0, 0.625, 1.25, 2.5, 3.75, 5, or 10 µmol/g of muscle) into pre-rigor bovine longissimus thoracis et lumborum (LTL) muscle samples, and pH and color were evaluated over a 48 h period. Injection of IAA blunted muscle pH decline and lowered lightness (L*), redness (a*), and yellowness (b*) values (P ≤ 0.05) in a concentration dependent fashion. In a follow-up study, LTL muscle samples were injected with 5 µmol IAA/g of muscle to test whether IAA maintains its effect over a 336 h post-mortem storage period. In addition to inhibiting pH decline and decreasing color values, IAA increased LTL muscle water holding capacity (WHC) and firmness (P ≤ 0.05) throughout the 336 h post-mortem storage period. Collectively, these data suggest that pre-rigor injection of IAA generates beef with dark cutting-like characteristics.

Myosin heavy chain isoform and metabolic profile differ in beef steaks varying in tenderness.

Our objective was to investigate possible differences in muscle fiber characteristics of beef longissimus lumborum (LL) steaks varying in tenderness (very tender vs. intermediate tender). Therefore, the relative abundance of myosin heavy chain (MHC) isoforms and activity/abundance of several glycolytic and oxidative enzymes were compared between the two steak groups. Greater (P < 0.05) content of MHC type IIa (MHC-IIa) and activities of phosphofructokinase (PFK) and glycogen phosphorylase (GP) were observed in the very tender steaks. Conversely, intermediate tender steaks had greater (P < 0.05) contents of MHC type I (MHC-I) and succinate dehydrogenase (SDH) and greater citrate synthase (CS) activity. Increased tenderness in the very tender steaks was associated with greater (P < 0.05) proteolysis as evaluated by desmin and troponin-T degradation. Further, mitochondrial calcium uniporter (MCU) was lower (P < 0.05) in the very tender steaks than steaks of intermediate tenderness. Collectively, shifting muscle characteristics toward a more glycolytic type appears to positively impact postmortem proteolysis and tenderization.

Inhibition of mitochondrial calcium uniporter enhances postmortem proteolysis and tenderness in beef cattle.

The purpose of this study was to examine the role of mitochondria in postmortem calcium homeostasis and its effect on proteolysis and tenderness. We hypothesized that mitochondria buffer cytosolic calcium levels and delay the activation of calpain-1 and subsequently the development of meat tenderness. To test this hypothesis, pre-rigor bovine longissimus thoracis et lumborum muscle samples were injected with DS16570511 to inhibit mitochondrial calcium uptake. Free calcium, tenderness, texture profile analysis (TPA), calpain-1 activity, and proteolysis were evaluated over a 336 h aging period. Inhibition of mitochondrial calcium uptake increased (P < .0001) cytosolic calcium concentration and calpain-1 autolysis and activity at 24 h compared to control steaks. Further, tenderness and TPA at 168 and 336 h, calpastatin degradation at 24 h, and proteolysis at 168 h were all enhanced (P < .05) in the treated steaks. Collectively, these data indicate that inhibition of mitochondrial calcium uptake can enhance postmortem proteolysis and tenderization through an early activation of calpain-1.