Heat-induced gelation of myofibrillar proteins and myosin (0.6M; pH 6.0) from rabbit fast-and slow-twitch muscles was analyzed by thermal scanning rheometry. Proteins from slow-twitch muscle exhibited higher thermostability and lower gel strength than those from fast-twitch muscle. Purifying myosin from myofibrillar proteins changed heat-gelation profiles and generally increased gel rigidity at 80ЊC. However, the effect of some proteins on the gelation of myosin was muscle dependent. Complete elimination of actin decreased the heat-gelling ability of slow myosin and increased that of fast myosin. Also, elimination of C-protein led to a greater increase in rigidity of gels from slow myosin than from fast myosin. The heat-behavior of the different protein fractions was related to the degree and type of aggregation in the gel.
The effects of ionic strength on myofibrils and myosin from rabbit fasttwitch Psoas major (PM) and slow-twitch Semimembranosus proprius (SMp) muscles before and after heating were studied by electron microscopy and thermal scanning rheometry. The direct suspension of proteins in low ionic strength (0.2M KCl; pH 6.0) led to very weak gels, whereas a gradual lowering of the ionic strength (by dialysis against 0.2M KCl; pH 6.0) of 0.6M KCl protein solutions induced strand-type networks at low temperature and strong heat-induced gels. As shown by transmission and scanning electron micrographs, in low ionic strength, SMp myosins formed shorter filaments before heating and thinner and shorter structures in heat-induced gels, as well as a lower gel porosity than PM myosins.
Although activation of CYP3A4 is frequently observed in vitro, predictive computational-based models and methods for in vitro-in vivo scaling are scarce. It has been previously shown that in vitro CYP3A4 heteroactivation of carbamazepine (CBZ)-epoxide (ep) formation can be associated with the clinical drug interaction between felbatame and CBZ. The previously reported prediction methodology is applied here to an additional set of in vitro CYP3A4 heteroactivators, some exerting this effect at concentrations relevant in vivo. The antimalarial artemisinin potently increases CBZ-ep formation by a maximum of 500% at 300 M. Testosterone and progesterone activates by a maximum of 1680 and 920%, respectively, at 150 M, and quinidine causes a 130% increase at 300 M. The predicted maximum in vivo decrease in steady-state concentration of carbamazepine (Css CBZ ) at saturating effector concentrations is 85 to 90% for testosterone and progesterone, 75% for artemisinin, and 45% for quinidine. The corresponding predicted in vivo increase in Css CBZ-ep is 50, 60, 55, and 30% for artemisinin, testosterone, progesterone, and quinidine, respectively. At effector concentrations relevant in vivo, the Css CBZ change is predicted to Յ20% for testosterone, artemisinin, and quinidine and Յ10% for progesterone, with a concomitant Css CBZ-ep increase of 12% for testosterone and Յ10% for progesterone, artemisinin, and quinidine. Structure-heteroactivation relationships were evaluated by generating a pharmacophore. The model includes two hydrogen bond acceptor features separated by hydrophobic features. Internal predictivity is high, and heteroactivation of an external test set correlate to observed in vitro heteroactivation.Inhibition of cytochromes P450 is a well known phenomena in vitro and has been shown to translate to decreases in P450-mediated metabolism in vivo, sometimes leading to clinically relevant drug-drug interactions (Lin and Lu, 1998). Quantitative prediction of the in vivo relevance of P450 inhibition based on in vitro data has also become an integral part of drug discovery and is based on methods that are continuously validated by comparison with in vivo observations (Ito et al., 2004). An increased number of attempts have also been made at producing computational models of P450 inhibition (Afzelius et al., 2001;Ekins et al., 2001).Extensive observations, however, suggest that apart from inhibition, modulation of cytochromes P450 can also occur by activation, a phenomenon that if translated to in vivo events is also a potential source of drug-drug interactions. The in vivo effects of P450 heteroactivation would be observed as an increased clearance of the affected drug, similar to the changes observed by P450 induction, except that the time course of the interaction would not be determined by the rate of synthesis and turnover of P450 enzyme but by the halflives of the interacting drugs. Activation of CYP3A4 has been extensively studied in vitro and based on site-directed mutagenesis studies (Khan et al., 2002), ...
Cocaine has been associated with hepatotoxicities in man and is a potent hepatotoxin in mice. The theorized toxic metabolite of cocaine is thought to be generated by a multistep pathway mediated primarily by cytochrome P-450. Ethanol, whether administered acutely or chronically, is known to have diverse effects on numerous hepatocellular biochemical pathways. The present study was designed to characterize not only the effects of acute and chronic ethanol on cocaine-mediated hepatotoxicity but also on the hepatic reduced glutathione (GSH) in an attempt to correlate depletions of GSH with changes in toxicity. Male and female mice were administered an acute 50 mg/kg dose of cocaine either 1 hr after an acute 3 g/kg dose of ethanol, or after 5 days of consuming an ethanol-containing liquid diet. Serum alanine aminotransferase (ALT) activity was measured in blood collected 24 hr after the acute cocaine dose. In addition, hepatic reduced glutathione (GSH) and cytochrome P-450 content were measured at the point in the pretreatment where cocaine was administered. The results of this study indicate that both acute and chronic ethanol pretreatment can markedly enhance the hepatotoxicity of cocaine in both male and female mice and that the enhancement is significantly greater after chronic ethanol pretreatment. Hepatic GSH was slightly decreased 1 hr after an acute dose of ethanol and significantly decreased after chronic ethanol consumption.(ABSTRACT TRUNCATED AT 250 WORDS)
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.