• Food Science of Animal Resources
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• v.34 no.5
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• pp.656-664
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• 2014

The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscle by liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer (n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide queries were obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43 queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicate identity or extensive homology, p<0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowse score (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirm the expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8 queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

• BMB Reports
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• v.48 no.1
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• pp.48-53
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• 2015

Oxidized LDL (oxLDL) performs critical roles in atherosclerosis by inducing macrophage foam cell formation and promoting inflammation. There have been reports showing that oxLDL modulates macrophage cytoskeletal functions for oxLDL uptake and trapping, however, the precise mechanism has not been clearly elucidated. Our study examined the effect of oxLDL on non-muscle myosin heavy chain IIA (MHC-IIA) in macrophages. We demonstrated that oxLDL induces phosphorylation of MHC-IIA (Ser1917) in peritoneal macrophages from wild-type mice and THP-1, a human monocytic cell line, but not in macrophages deficient for CD36, a scavenger receptor for oxLDL. Protein kinase C (PKC) inhibitor-treated macrophages did not undergo the oxLDL-induced MHC-IIA phosphorylation. Our immunoprecipitation revealed that oxLDL increased physical association between PKC and MHC-IIA, supporting the role of PKC in this process. We conclude that oxLDL via CD36 induces PKC-mediated MHC-IIA (Ser1917) phosphorylation and this may affect oxLDL-induced functions of macrophages involved in atherosclerosis.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.1
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• pp.11-21
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• 2009

Objectives : This study was performed to investigate the effects of Semen Persicae(SP) on muscle type change and vascularization effect by measuring expression of Myosin heavy chain (MHC) and Vascular endothelial growth factor (VEGF) protein in the Middle cerebral artery occlusion(MCAo) rats. Methods : The middle cerebral artery was occluded, SP extraction was administrated for 4 days. The effects of SP on muscle type change and vascularization were measured. Results : 1. VEGF protein expressions in the Semen Persicae oral administration group of MCAo group were increased compared to the control group. 2. There were no significant difference between the Semen Persicae oral administration of MCAo group and the control group in MHC isoform (Type I, Type IIa) expression change. Conclusions : The present study demonstrates the effect of Semen Persicae in the vascularizing after ischemia, but has no significant effect in musle type change and the improvement of musle atrophy.

• Animal cells and systems
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• v.14 no.1
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• pp.25-30
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• 2010

We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

• Food Science of Animal Resources
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• v.36 no.6
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• pp.819-828
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• 2016

Muscle fiber characteristics account for meat quality and muscle fibers are mainly classified into three or more types according to their contractile and metabolic properties. However, the majority of previous studies on bovine skeletal muscle are based on myosin ATPase activity. In the present study, the differences in the characteristics of muscle fibers classified by the expression of myosin heavy chain (MHC) among four bovine skeletal muscles such as longissimus thoracis (LT), psoas major (PM), semimembranosus (SM) and semi-tendinosus (ST) and their relationships to beef quality were investigated. MHCs 2x, 2a and slow were identified by LC-MS/MS and IIX, IIA and I fiber types were classified. PM, which had the smallest size and highest density of fibers regardless of type, showed the highest myoglobin content, CIE $L^*$, $a^*$, $b^*$ and sarcomere length (p<0.05), whereas ST with the highest composition of IIX, showed high shear force and low sarcomere length (p<0.05). The correlation coefficients between muscle fiber characteristics and meat quality showed that type IIX is closely related to poor beef quality and that a high density of small-sized fibers is related to redness and tenderness. Therefore, the differences in meat quality between muscles can be explained by the differences in muscle fiber characteristics, and especially, the muscles with good quality are composed of more small-sized fibers regardless of fiber type.

• Food Science of Animal Resources
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• v.40 no.1
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• pp.132-144
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• 2020

The aim of this study was to optimize staining procedures for muscle fiber typing efficiently and rapidly in bovine and porcine skeletal muscles, such as longissimus thoracis, psoas major, semimembranosus, and semitendinosus muscles. The commercially available monoclonal anti-myosin heavy chain (MHC) antibodies and fluorescent dye-conjugated secondary antibodies were applied to immunofluorescence histology. Two different procedures, such as cocktail and serial staining, were adopted to immunofluorescence analysis. In bovine muscles, three pure types (I, IIA, and IIX) and one hybrid type, IIA+IIX, were identified by the cocktail procedure with a combination of BA-F8, SC-71, BF-35, and 6H1 anti-MHC antibodies. Porcine muscle fibers were typed into four pure types (I, IIA, IIX, and IIB) and two hybrid types (IIA+IIX and IIX+IIB) by a serial procedure with a combination of BA-F8, SC-71, BF-35, and BF-F3. Unlike for bovine muscle, the cocktail procedure was not recommended in porcine muscle fiber typing because of the abnormal reactivity of SC-71 antibody under cocktail procedure. Within the four antibodies, combinations of two or more anti-MHC antibodies allowed us to distinguish pure fiber types or all fiber types including hybrid types. Application of other secondary antibodies conjugated with different fluorescent dyes allowed us to get improved image resolution that clearly distinguished hybrid fibers. Muscle fiber characteristics differed depending on species and muscle types.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.640-645
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• 1990

Changes of the electrophoretic patterns of myofibrillar proteins by sugar audition and heat treatment was studied. In the electrophoretic patterns of myofibrills prepared from no sugar added meat, as the intensity of higher molecular weight band such as myosin heavy chain showed a remarkable decrease by heating, that of lower molecular weight band such as actin showed no change. That from sugar added meat showed more remarkable decrease in the intensity of higher molecular weight band than that from no sugar added meat and this tendency was most noticeable in case of glucose addition. The effect of digestion with proteases after sugar addition and heat treatment on the electrophoretic patterns exhibited the descending order of trypsin >chymotrypsin >peptidase. By digestion with these three enzymes at one time myosin produced 27.000 dalton and 32.000 dalton components, and actin showed 16,000 dalton component. in the case of heat treatment, a part of actin was not digested. And in the case of glucose addition the myosin aggregates was not digested with these three enzymes at a time.

• Korean Journal of Food Science and Technology
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• v.28 no.2
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• pp.292-297
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• 1996

In this study, biochemical changes of muscle protein in squid Sikhae prepared at various temperatures and moisture content were investigated. The pH values in squid Sikhae decreased rapidly with increase in titratable acidity during fermentation at high temperature and moisture content. Growth rate of lactic acid bacteria increased rapidly during fermentation at high temperature and moisture content. Changes in muscle protein subunit were analyzed by SDS-PAGE; myosin heavy chain component disappeared completely during preparation of squid Sikhae and actin component decreased slowly during fermentation. Therefore, it was suggested that myosin heavy chain component in squid muscle protein was autolyzed at the incipient for-mentation time and actin component was degraded by acidic protease in muscle and protease produced from microorganisms during fermentation.

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.158-158
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• 2015

Myoblast are myogenic precursors that proliferate, activate, and differentiate on muscle injury to sustain the regenerative capacity of skeletal muscle; The neuronal isoform of nitric oxide synthase (nNOS, termed also NOS-I) is expressed in normal adult skeletal muscle, suggesting important functions for Nitric oxide (NO) in muscle biology1,2,3. However, the expression and subcellular localization of NO in muscle development and myoblast differentiation are largely unknown. In this study, we examined effects of the nitric oxide generated by a microwave plasma torch, on proliferation/differentiation of rat myoblastic L6 cells. Experimental data pertaining to nitric oxide production are presented in terms of the oxygen input in units of cubic centimetres per minute. The various levels of nitric oxide are observed depending on the flow rate of nitrogen gas, the ratio of oxygen gas, and the microwave power4. In order to evaluate the potential of nitric oxide as an activator of cell differentiation, we applied nitric oxide generated from the microwave plasma torch to L6 skeletal muscles. Differentiation of L6 cells into myotubes was significantly enhanced the differentiation after nitric oxide treatment. Nitric oxide treatment also increase the expression of myogenesis marker proteins and mRNA level, such as myogenin and myosin heavy chain (MHC), as well as cyclic guanosine monophosphate (cGMP), However during the myotube differentiation we found that NO activate oxidative stress signaling erks expression. Therefore, these results establish a role of NO and cGMP in regulating myoblast differentiation and elucidate their mechanism of action, providing a direct link with oxidative stress signalling, which is a key player in myogenesis. Based on these findings, nitric oxide generated by plasma can be used as a possible activator of cell differentiation and tissue regeneration.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.1
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• pp.102-108
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• 2006

Microbial transglutaminase (MTGase) was investigated to determine whether it was an effective binding agent for the processing of low-quality pork loins. MTGase especially promoted the coagulation of myosin heavy chain (MHC). However, the effect of MTGase on MHC from low-quality meat was less than that from the normal meat when the reaction time was not enough. The breaking strength of the heat-induced gel made of myosin B from low-quality meat with MTGase was lower than that of normal meat. Sausage made with low-quality meat with MTGase did not exhibit improved hardness, as compared to that made with normal meat. Results of this study indicated that use of low-quality meat in the manufacture of sausage was feasible to get textural property equal to that of normal meat sausage, when a half or more of the raw material was normal meat and MTGase was used in the sausage.