Influence of early postmortem protein oxidation on beef quality

The objective of this study was to examine the effect of early postmortem protein oxidation on the color and tenderness of beef steaks. To obtain a range of oxidation levels, the longissimus lumborum muscles (LM) from both strip loins of 20 steers fed either a finishing diet with vitamin E (1,000 IU per steer daily, minimum of 126 d [VITE]; n = 10 steers) or fed the same finishing diet without vitamin E (CON; n = 10 steers) were used. Within 24 h after slaughter, the LM muscle from each carcass was cut into 2.54-cm-thick steaks and individually vacuum packaged. Steaks from each steer were assigned to a nonirradiated group or an irradiated group. Steaks were irradiated within 26 h postmortem, and were aged at 4°C for 0, 1, 3, 7, and 14 d after irradiation. Steaks from each diet/irradiation/aging time treatment were used to determine color, shear force, and degree of protein oxidation (carbonyl content). Steaks from steers fed the VITE diet had higher (P \u3c 0.01) α-tocopherol contents than steaks from steers fed the CON diet. Immediately following irradiation, steaks that had been irradiated had lower (P \u3c 0.05) L* values regardless of diet. Irradiated steaks, regardless of diet, had lower a* (P \u3c 0.05) and b* (P \u3c 0.01) values than nonirradiated steaks at all aging times. Carbonyl concentration was higher (P \u3c 0.05) in proteins from irradiated steaks compared to nonirradiated steaks at 0, 1, 3, and 7 d postirradiation. Immunoblot analysis showed that vitamin E supplementation decreased the number and extent of oxidized sarcoplasmic proteins. Protein carbonyl content was positively correlated with Warner-Bratzler shear force values. These results indicate that increased oxidation of muscle proteins early postmortem could have negative effects on fresh meat color and tenderness

Effect of pH and ionic strength on μ- and m-calpain inhibition by calpastatin

The objectives of this study were to determine the extent to which pH and ionic strength influence μ- and m-calpain activity and the inhibition of calpains by calpastatin. Calpastatin, μ-calpain, and m-calpain were purified from at-death porcine semimembranosus. μ-Calpain or m-calpain (0.45 U) were incubated with the calpain substrate Suc-Leu-Leu-Val-Tyr-7-amino-4-methyl coumarin in the presence of calpastatin (0, 0.15, or 0.30 U of calpain inhibitory activity) under the following pH and ionic strength conditions: pH 7.5 and 165 mM NaCl or 295 mM NaCl; pH 6.5 and 165 mM NaCl or 295 mM NaCl; and pH 6.0 and 165 mM NaCl or 295 mM NaCl. The reactions were initiated with addition of 100 μM (μ-calpain) or 1 mM CaCl2 (m-calpain), and calpain activity was recorded at 30 and 60 min. μ-Calpain had the greatest (P \u3c 0.01) activity at pH 6.5 at each ionic strength. Higher ionic strength decreased μ-calpain activity (P\u3c 0.01) at all pH conditions. Inhibition percent of μ-calpain by calpastatin was not affected by pH; however, it was influenced by ionic strength. Inhibition of μ-calpain by calpastatin was higher (P \u3c 0.01) at 295 mM NaCl than at 165 mM NaCl when 0.3 units of calpastatin were included in the assay. Activity of m-calpain was greater (P \u3c 0.01) at pH 7.5 than at pH 6.5. m-Calpain activity was not detected at pH 6.0. Inhibition of m-calpain was greater (P \u3c 0.01) when 0.15 and 0.3 U calpastatin were added at pH 6.5 than 7.5 at 165 mM NaCl, whereas percentage inhibition of m-calpain was greater (P \u3c 0.01) at 295 mM than 165 mM NaCl at pH 7.5 and 6.5. These observations provide new evidence that defines further the influence of pH decline and increased ionic strength on μ-calpain, m-calpain, and calpastatin activity, thereby helping to more accurately define a role for these enzymes in the process of postmortem tenderization

Oxidative environments decrease tenderization of beef steaks through inactivation of μ-calpain

This study was designed to test the hypothesis that oxidative conditions in postmortem (PM) tissue decrease calpain activity and proteolysis, subsequently minimizing the extent of tenderization. To achieve different levels of oxidation, the diets of beef cattle were supplemented with vitamin E for the last 126 d on feed, and beef steaks were irradiated early PM. Ten steers were fed a finishing diet with the inclusion of vitamin E at 1,000 IU per steer daily (VITE). Another 10 beef steers were fed the same finishing diet without added vitamin E (CON). At 22 to 24 h PM, strip loins from each carcass were cut into 2.54-cm-thick steaks and individually vacuum packaged. Within 26 h PM, steaks were irradiated at 0 or 6.4 kGy and then aged at 4°C for 0, 1, 3, 7, and 14 d postirradiation. Steaks from each time point were used to determine Warner-Bratzler shear force (WBSF) and calpain activity, and for western blotting of sarcoplasmic proteins and myofibrillar proteins. Calpastatin activity was determined at 0, 3, and 14 d postirradiation. At 1, 3, 7, and 14 d postirradiation, WBSF values of irradiated steaks were higher (P \u3c 0.03) than for nonirradiated steaks. Western blots of troponin-T and desmin showed decreased proteolysis in irradiated samples compared with nonirradiated samples. At 2 d PM, troponin-T degradation products were more evident (P \u3c 0.03) in nonirradiated steaks supplemented with VITE than nonirradiated steaks from the CON diet. Similarly, VITE treatment resulted in steaks with lower (P \u3c 0.05) calpastatin activity at 1 d PM than in steaks from steers fed the CON diet. Irradiation diminished the rate of calpastatin inactivation. Irradiated samples, regardless of diet, had no detectable levels of intact titin or nebulin. Irradiation decreased μ-calpain activity and autolysis, whereas m-calpain activity was not affected by diet or irradiation. Inactivation of μ-calpain by oxidation during early times PM decreased the amount of myofibrillar proteolysis, thereby decreasing the extent of tenderization of beef steaks

Effect of oxidation, pH, and ionic strength on calpastatin inhibition of μ- and m-calpain

The objective of this study was to evaluate the effect of oxidation on μ- and m-calpain activity at varying pH and ionic strength conditions in the presence of calpastatin. In 2 separate experiments, purified porcine skeletal muscle μ- or m-calpain (0.45 units of caseinolytic activity) was incubated in the presence of calpastatin (0, 0.15, or 0.30 units) at pH 7.5, 6.5, or 6.0 with either 165 or 295 mM NaCl. The reactions were initiated with the addition of CaCl2 (100 μM for μ-calpain; 1 mM for m-calpain). In Experiment 1, μ- or m-calpain was incubated with the calpain substrate Suc-Leu-Leu-Val-Tyr-AMC (170 μM). Either 0 or 16 μ μM H2O2 was added to each assay. Activity was measured at 60 min. In Experiment 2, calpain was incubated with highly purified porcine myofibrils (4 mg/mL) under conditions described. Either 0 or 100 μM H2O2 was added immediately prior to the addition of calpain. Degradation of desmin was determined on samples collected at 2, 15, 60, and 120 min. Results from Experiment 1 indicated that oxidation decreased (P \u3c 0.01) activity of μ-calpain. μ-Calpain had the greatest (P \u3c 0.01) activity at pH 6.5, and m-calpain had the greatest (P \u3c 0.01) activity at pH 7.5 at 60 min. m-Calpain activity was not detected at pH 6.0. μ- and m-calpain activity were lower (P \u3c 0.01) at 295 mM NaCl than at 165 mM NaCl at all pH conditions. Oxidation lowered (P \u3c 0.01) calpastatin inhibition of μ-and m-calpain at all pH and ionic strength combinations. In Experiment 2, oxidation decreased proteolytic activity of μ-calpain against desmin at pH 6.0 (P \u3c 0.05 at 15, 60, and 120 min) and decreased m-calpain at all pH conditions. However, desmin degradation by μ-calpain was not as efficiently inhibited by calpastatin at pH 7.5 and as at pH 6.5 (P = 0.03 at 60 min) when oxidizing conditions were created. This is consistent with the results from Experiment 1, which indicated that oxidation decreased the ability of calpastatin to inhibit μ-calpain. These studies provide evidence that oxidation influences calpain activity and inhibition of calpains by calpastatin differently under varying environmental conditions. The results suggest that, at the higher pH conditions used, calpastatin may limit the possibility of oxidation-induced inactivation of μ-calpain

The Energy Conversion Playground (ECP) Design Task: Assessing How Students Think About Technical and Non-technical Considerations in Sustainable Community Development

Students in global service-learning and similar programs frequently encounter substantial social, cultural, political, and ethical differences when working with project partners in different countries and regions. Neglecting such differences can lead to project failures and/or disempowered communities. In response to these challenges, educational resources have been developed to teach students to think about how the people, social structures, and other contextual factors associated with projects can affect, and be affected by, students’ designs. Yet, there remains a scarcity of valid and reliable instruments to evaluate the effectiveness of such interventions. The purpose of this study is create a theoretically and empirically grounded instrument, the Energy Conversion Playground (ECP) design task, that is able to provide a meaningful and robust assessment of an individual’s ability to identify salient technical and non-technical considerations when approaching an engineering design task situated in a developing country context. We present the scenario and an accompanying rubric that was first developed inductively from student responses to the scenario (specifically 449 discrete items from 93 ECP design tasks submitted by students who attended a Global Engineering Design Symposium). Further development of the rubric involved deductive grounding in relevant literature. To demonstrate the sensitivity of ECP design task to changes in students’ thinking, we also performed comparative analysis of responses from a subset of the students (n=37) who completed the same instrument both before and after participating in the GEDS

MDMA Decreases Gluatamic Acid Decarboxylase (GAD) 67-Immunoreactive Neurons in the Hippocampus and Increases Seizure Susceptibility: Role for Glutamate

3,4-Methylenedioxy-methamphetamine (MDMA) is a unique psychostimulant that continues to be a popular drug of abuse. It has been well documented that MDMA reduces markers of 5-HT axon terminals in rodents, as well as humans. A loss of parvalbumin-immunoreactive (IR) interneurons in the hippocampus following MDMA treatment has only been documented recently. In the present study, we tested the hypothesis that MDMA reduces glutamic acid decarboxylase (GAD) 67-IR, another biochemical marker of GABA neurons, in the hippocampus and that this reduction in GAD67-IR neurons and an accompanying increase in seizure susceptibility involve glutamate receptor activation. Repeated exposure to MDMA (3×10mg/kg, ip) resulted in a reduction of 37–58% of GAD67-IR cells in the dentate gyrus (DG), CA1, and CA3 regions, as well as an increased susceptibility to kainic acid-induced seizures, both of which persisted for at least 30 days following MDMA treatment. Administration of the NMDA antagonist MK-801 or the glutamate transporter type 1 (GLT-1) inducer ceftriaxone prevented both the MDMA-induced loss of GAD67-IR neurons and the increased vulnerability to kainic acid-induced seizures. The MDMA-induced increase in the extracellular concentration of glutamate in the hippocampus was significantly diminished in rats treated with ceftriaxone, thereby implicating a glutamatergic mechanism in the neuroprotective effects of ceftriaxone. In summary, the present findings support a role for increased extracellular glutamate and NMDA receptor activation in the MDMA-induced loss of hippocampal GAD67-IR neurons and the subsequent increased susceptibility to evoked seizures

Me And Mrs. Jones

Illustration of man and woman\u27s silhouettehttps://scholarsjunction.msstate.edu/cht-sheet-music/6495/thumbnail.jp

Simulations of the OzDES AGN Reverberation Mapping Project

As part of the OzDES spectroscopic survey we are carrying out a large scale reverberation mapping study of $\sim$500 quasars over five years in the 30 deg$^2$ area of the Dark Energy Survey (DES) supernova fields. These quasars have redshifts ranging up to 4 and have apparent AB magnitudes between $16.8<r<22.5$ mag. The aim of the survey is to measure time lags between fluctuations in the quasar continuum and broad emission line fluxes of individual objects in order to measure black hole masses for a broad range of AGN and constrain the radius-luminosity ($R-L$) relationship. Here we investigate the expected efficiency of the OzDES reverberation mapping campaign and its possible extensions. We expect to recover lags for $\sim$35-45\% of the quasars. AGN with shorter lags and greater variability are more likely to yield a lag, and objects with lags $\lesssim$6 months or $\sim$1 year are expected be recovered the most accurately. The baseline OzDES reverberation mapping campaign is predicted to produce an unbiased measurement of the $R-L$ relationship parameters for H$\beta$, Mg II $\lambda$2798, and C IV $\lambda$1549. However, extending the baseline survey by either increasing the spectroscopic cadence, extending the survey season, or improving the emission line flux measurement accuracy will significantly improve the $R-L$ parameter constraints for all broad emission lines.Comment: Published online in MNRAS. 28 page

Effect of pectin, lecithin, and antacid feed supplements (Egusin®) on gastric ulcer scores, gastric fluid pH and blood gas values in horses

BACKGROUND: The objectives of this study were to evaluate the effects of two commercial feed supplements, Egusin 250(®) [E-250] and Egusin SLH(®) [E-SLH], on gastric ulcer scores, gastric fluid pH, and blood gas values in stall-confined horses undergoing feed-deprivation. METHODS: Nine Thoroughbred horses were used in a three-period crossover study. For the three treatment groups, sweet feed was mixed with E-250, E-SLH, or nothing (control group) and fed twice daily. Horses were treated for 21 days, then an additional 7 days while on an alternating feed-deprivation model to induce or worsen ulcers (period one). In periods two and three, horses (n=6) were treated for an additional 7 days after feed-deprivation. Gastroscopies were performed on day -1 (n=9), day 21 (n=9), day 28 (n=9) and day 35 (n=6). Gastric juice pH was measured and gastric ulcer scores were assigned. Venous blood gas values were also measured. RESULTS: Gastric ulcers in control horses significantly decreased after 21 days, but there was no difference in ulcer scores when compared to the Egusin® treated horses. NG gastric ulcer scores significantly increased in E-250 and control horses on day 28 compared to day 21 as a result of intermittent feed-deprivation, but no treatment effect was observed. NG ulcer scores remained high in the control group but significantly decreased in the E-SLH- and E-250-treated horses by day 35. Gastric juice pH values were low and variable and no treatment effect was observed. Mean blood pCO(2) values were significantly increased two hours after feeding in treated horses compared to controls, whereas mean blood TCO(2) values increased in the 24 hour sample, but did not exceed 38 mmol/l. CONCLUSIONS: The feed-deprivation model increased NG gastric ulcer severity in the horses. However, by day 35, Egusin(®) treated horses had less severe NG gastric ulcers compared to untreated control horses. After 35 days, Egusin(®) products tested here ameliorate the severity of gastric ulcers in stall-confined horses after feed stress