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Whole-body phenylalanine kinetics and skeletal muscle protein signaling in horses with pituitary pars intermedia dysfunction
2014
Mastro, Laurel M. | Adams, Amanda A. | Urschel, Kristine L.
Objective—To compare whole-body phenylalanine kinetics and the abundance of factors in signaling pathways associated with skeletal muscle protein synthesis and protein breakdown between horses with pituitary pars intermedia dysfunction (PPID) and age-matched control horses without PPID. Animals—12 aged horses (6 horses with PPID and 6 control horses; mean age, 25.0 and 25.7 years, respectively). Procedures—Plasma glucose, insulin, and amino acids concentrations were determined before and 90 minutes after feeding. Gluteal muscle biopsy samples were obtained from horses 90 minutes after feeding, and the abundance and activation of factors involved in signaling pathways of muscle protein synthesis and breakdown were determined. The next day, horses received a priming dose and 2 hours of a constant rate infusion of 13C sodium bicarbonate followed by a priming dose and 4 hours of a constant rate infusion of 1-13C phenylalanine IV; whole-body protein synthesis was determined. Results—Plasma glucose and insulin concentrations were higher after feeding than they were before feeding for both groups of horses; however, no significant postprandial increase in plasma amino acids concentrations was detected for either group. Phenylalanine flux, oxidation, release from protein breakdown, and nonoxidative disposal were not significantly different between groups. No significant effect of PPID status was detected on the abundance or activation of positive or negative regulators of protein synthesis or positive regulators of protein breakdown. Conclusions and Clinical Relevance—Results of this study suggested that whole-body phenylalanine kinetics and the postprandial activation of signaling pathways that regulate protein synthesis and breakdown in muscles were not affected by PPID status alone in aged horses.
Show more [+] Less [-]Effect of small interfering RNAs on in vitro replication and gene expression of feline coronavirus
2014
Anis, Eman A. | Wilkes, Rebecca P. | Kania, Stephen A. | Legendre, Alfred M. | Kennedy, Melissa A.
Objective—To evaluate the ability of small interfering RNAs (siRNAs) to inhibit in vitro viral replication and gene expression of feline coronavirus (FCoV). Sample—Cell cultures of Crandell-Rees feline kidney cells. Procedures—5 synthetic siRNAs that each targeted a different region of the FCoV genome were tested individually and in various combinations for their antiviral effects against 2 strains of FCoV (feline infectious peritonitis virus WSU 79-1146 and feline enteric coronavirus WSU 79-1683) in cell cultures. Tested combinations targeted the FCoV leader and 3′ untranslated region, FCoV leader region and nucleocapsid gene, and FCoV leader region, 3′ untranslated region, and nucleocapsid gene. For each test condition, assessments included relative quantification of the inhibition of intracellular viral genomic RNA synthesis by means of real-time, reverse-transcription PCR analysis; flow cytometric evaluation of the reduction of viral protein expression in infected cells; and assessment of virus replication inhibition via titration of extracellular virus with a TCID50 infectivity assay. Results—The 5 siRNAs had variable inhibitory effects on FCoV when used singly. Combinations of siRNAs that targeted different regions of the viral genome resulted in more effective viral inhibition than did individual siRNAs that targeted a single gene. The tested siRNA combinations resulted in approximately 95% reduction in viral replication (based on virus titration results), compared with findings in negative control, nontargeting siRNA–treated, FCoV-infected cells. Conclusions and Clinical Relevance—In vitro replication of FCoV was specifically inhibited by siRNAs that targeted coding and noncoding regions of the viral genome, suggesting a potential therapeutic application of RNA interference in treatment of feline infectious peritonitis.
Show more [+] Less [-]In vitro digestibility study of fractionated Refined, Bleached and Deodorized Palm Stearin
2014
Shawaluddin T. | Zahariah I. | Mardhati M. | Norliza S. | Mohd Suria A. Y. | Wong H. K.
In vitro digestibility study was conducted to determine the effectsof supplementing fractionated Refined,Bleached and Deodorized Palm Stearin(RBDPST) on ruminal digestion. Fractionated RBDPST was soaked in incubation medium consisting of distilled water, buffer solution, trace element solution, micro and macro mineral solution, as well as rumen liquor that was collected from slaughtered cattle. This experiment was conducted at 39°C with an incubation period of 24 hours. Dried napier grass was used as control treatment. Gas producedwas recorded and collected to measure the methane gas produced. Methane gas produced from fractionated RBDPST was found to be relatively lower than control. This indicates that fractionated RBDPST had the ability to function as rumen bypass fat as it was not fully digested in the rumen.
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