Comparision of staining methods for two dimensional electrophoresis gel resolved with Puntius javanicus liver proteome

The aim of this study was to compare the various staining methods based on commassie briliant blue and silver nitrate stain for the two dimensional gel electrophoresis resolved with Puntius javanicus liver proteome. The staining methods were selected base on the previous reportabout their compatiblitywiththe mass spectrometry analysis. Sliver staining methodis known as the most sensitive method to visualize the maximum number of protein spots resolved in 2D gel but it is less sensitive(incompatible) toward mass spectrometry detection. Results of this study showed that a modifiedstaining method using colloidal coomassie blue G-250 (CCB) is roughly similarly sensitive but lower protein spot detected compared with silver staining (SS) as indicated at the number of 303±26 and 693±14of proteinresolved in both types of stained gels. The conventional methods of staining using commassie brilliant blue G-250 and R-250 only detected less number of protein spots(128±17and 78±11, respectively) compared to modified CCB staining method. As the commasie brilliant blue stain is known to be a very sensitive for mass spectrometry detection, the modified method of CCB was selected for further study on Puntius javanicus liver proteome

A proteomic based assessment on changes in myofirbrillar proteins of goat longissimus muscle as affected by heat treatments

The present study examined the effect of different heat treatments; (1) chilled, (2) boiled at 100°C for 30 min, and (3) autoclaved at 121°C at 15 psi for 20 min, on the expression of goat skeletal muscle proteins using two-dimensional gel electrophoresis. The molecular weight (MW) and isoelectric point (pI) of heat stable proteins were characterised followed by identification of the proteins by MALDI-TOF/TOF mass spectrometry. There were 153 protein spots obtained in the boiled samples, while only 46 protein spots were observed in the autoclaved samples. Thirteen spots that exhibited high intensity of protein were chosen from the autoclaved sample for MALDI-TOF/TOF mass spectrometry analysis. The putative heat stable proteins identified were myosin light chain (MLC), actin, tropomyosin (TPM), troponin T (TnT), myoglobin, and creatine kinase. The Proc-GLM analysis revealed that the heat treatments have resulted in significant differences in spot intensities of actin, troponin T (TnT), myoglobin, and creatine kinase with no significant changes noted in other proteins

Skeletal muscle proteome and meat quality of broiler chickens subjected to gas stunning prior slaughter or slaughtered without stunning

The study examined the effects of pre-slaughter gas stunning and slaughter without stunning on meat quality and skeletal muscle proteome of broiler chickens. Fifty Cobb broiler chickens were randomly assigned to either a neck cut without pre-slaughter stunning (Halal slaughter) or pre-slaughter gas stunning followed by a neck cut. Samples of Pectoralis major muscle at 7 min, 4 h and 24 h postmortem were analyzed for pH, shear force, color, drip and cooking losses. Proteome profile of the 7 min samples was examined by two-dimensional polyacrylamide gel electrophoresis. Birds subjected to Halal slaughter had higher (P < 0.05) redness than those gas stunned at 4 and 24 h postmortem. Gas-stunned birds had lower (P < 0.05) muscle pH and shear force and higher (P < 0.05) drip and cooking losses compared with those subjected to Halal slaughter throughout postmortem storage. Gas stunning up-regulated (P < 0.05) the expression of beta-enolase, pyruvate kinase and creatine kinase compared with Halal slaughter. Results indicate that pre-slaughter gas stunning hastened postmortem energy metabolism and had detrimental effects on the water holding capacity and redness of broiler breast muscles

Transforming agriculture research into commercialisation: experience of Universiti Putra Malaysia

One of the major goals of any high impact research and development is an overall improvement in the well-being and sustainable quality of life through innovations. As universities continuously disseminate innovations from R&D activities, many prototypes and lab-scale products, whether tangible or intangible, can be made available for public use. The success of bringing these innovations to the marketplace depends on the quality and capability of the technology transfer office to lead different types of activities, engagements, negotiation and inclusiveness towards fulfilling the needs of commercialisation partners and the market. This paper presented a general overview of transforming research output into commercialisation in the context of Universiti Putra Malaysia (UPM). Throughout this paper, different commercialization channels, the roles of technology transfer offices and multiple agencies are further discussed with a special focus on agricultural innovations and technologies. This review contributes to both academic and agricultural industry research, development and commercialization activities by illustrating current innovation produced by UPM and industry-university collaboration, conducted at a leading agriculture university

LC–QTOF-MS identification of porcine-specific peptide in heat treated pork identifies candidate markers for meat species determination

The purpose of this study was to identify porcine-specific peptide markers from thermally processed meat that could differentiate pork from beef, chevon and chicken meat. In the initial stage, markers from tryptic digested protein of chilled, boiled and autoclaved pork were identified using LC–QTOF-MS. An MRM method was then established for verification. A thorough investigation of LC–QTOF-MS data showed that only seven porcine-specific peptides were consistently detected. Among these peptides, two were derived from lactate dehydrogenase, one from creatine kinase, and four from serum albumin protein. However, MRM could only detect four peptides (EVTEFAK, LVVITAGAR, FVIER and TVLGNFAAFVQK) that were consistently present in pork samples. In conclusion, meat species determination through a tandem mass spectrometry platform shows high potential in providing scientifically valid and reliable results even at peptide level. Besides, the specificity and selectivity offered by the proteomics approach also provide a robust platform for Halal authentication

Comparing the effect of heat on tropomyosin isoforms patterns from water buffalo and wild boar meat by two-dimensional gel electrophoresis

Tropomyosin is one of the most abundant proteins in meat; however, very little is known about it due to the lack of scientific literature. In this study, the spot volume of tropomyosin (TPM) isoforms, TPM2 and TPM1, in meat from water buffalo and wild boar subjected to various cook treatments were compared. We hypothesized that primary structures of the tropomyosin isoforms from both species would remain stable despite the application of heat. Proteins extracted from the treated meats were analyzed using two-dimensional gel electrophoresis and mass spectrometry. A Kruskal-Wallis test showed that there were no significant differences in protein spot volumes for all treatments; however, a significant difference was observed between species. Changes in the amino acid sequence of TPM1 were observed between the two species, indicating that the isoforms could be used as thermostable proteins or peptide markers for species identification because of their resistance to high temperatures

Cytokines (IL 1β and IL 6) responses in non-pregnant does infected with corynebacterium pseudotuberculosis following intradermal route of infection in chronic state

Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis (CLA) which commonly affects sheep and goats. The disease remains as a major disease causing economic loss to the small ruminant industries. There is little information related to responses of interleukin-1β and interleukin-6 in the chronic states. This study was designed to determine the serum concentrations of interleukin-1β and interleukin-6 (pg/mL) in non-pregnant does experimentally inoculated with Corynebacterium pseudotuberculosis via intradermal route in chronic form. Eighteen non-pregnant healthy Katjang does aged 2 years old were divided randomly into two groups. The control and the treatment groups consist of nine does each and were kept for 3 months. The control group was inoculated with PBS solution while the treatment group was inoculated intradermally with C. pseudotuberculosis. Serum samples were collected every 3 days (72 hours) for 3 months (2064 hours). The present study showed significant increase in IL-1β (278 ± 19.19 pg/mL) after 1day (24 hours) of post infection (p<0.0001) which decreased sharply (98.31 ± 19.19 pg/mL) after 5 days (120 hours) of post infection (p = 0.9293) and attained a significant concentration (217.43 ± 19.19 pg/mL) after 3 months (2064 hours) of post infection (p<0.0048) in does challenged with C. pseudotuberculosis compared to the control group. In contrast, the concentration of IL-6 increased significantly (p<0.0001) to (56.43 ± 1.98 pg/mL) in 2 months (1392 hours) of post infection and then decreased significantly (p<0.0001) to concentration of (22.18 ± 1.98 pg/mL) in 3 months (2064 hours) compared to the control group. In conclusion, the present study indicate that the importunity of C. pseudotuberculosis is associated with persistently high concentrations of IL-1β and low concentration of IL-6 which, when interpreted, could severely contribute to pathological vicissitudes and injury of organs and tissues in the chronic stage of C. pseudotuberculosis infections