• Asian-Australasian Journal of Animal Sciences
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• v.18 no.3
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• pp.427-432
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• 2005

The purpose of this study was performed as model study using four animals to investigate the correction between the changes in Differential Scanning Calorimetry thermogram of muscle proteins during storage and meat freshness. M. longissimus dorsi of pork was obtained immediately after slaughter and chilled/stored at either $-2^{\circ}C$ or $25^{\circ}C$ for up to 96 h for analyses. DSC thermograms were determined and compared with pH values, ATP-related compounds, K-values, volatile basic nitrogen (VBN) levels, bacterial counts and electrophoretic behavior. Changes in pH, bacterial counts, VBN and K-values were associated with increased storage temperature and time. The levels of pH values, bacterial counts, VBN and K-values of pork samples stored at $25^{\circ}C$ were higher than those of the pork samples stored at $-2^{\circ}C$. ATP concentration decreased faster in samples stored at $25^{\circ}C$. Only IMP increased in samples stored at $-2^{\circ}C$, whereas the concentration of hypoxanthine and inosine increased in samples stored at $25^{\circ}C$. One exothermic peak and two endothermic peaks appeared on the thermograms of pork stored at either temperature. Lower transition temperature of myosin, sarcoplasmic protein and actin peaks were observed. The freshness parameters of K-value, VBN and hypoxanthine showed highly negative correlations (-0.742- -0.9980) to the changes in transition temperature. Therefore, the shift temperature on DSC thermogram can be used as an indicator of the freshness parameters of meat.

• Genomics & Informatics
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• v.5 no.2
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• pp.77-82
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• 2007

Serotonin (5-HT), the endogenous nonselective 5-HT receptor agonist, activates the inositol-1,4,5-triphosphate/calcium $(InsP3/Ca^{2+})$ signaling pathway and exerts both stimulatory and inhibitory actions on cAMP production and neuromodulin secretion in rat hypothalamic neurons. Specific mRNA transcripts for 5-HT1A, 5-HT2C and 5-HT4 were identified in rat hypothalamic neurons. These experiments were supported by combined techniques such as cAMP and a $Ca^{2+}$ assays in order to elucidate the associated receptors and signaling pathways. The cAMP production and neuromodulin release were profoundly inhibited during the activation of the Gi-coupled 5-HT1A receptor. Treatment with a selective agonist to activate the Gq-coupled 5-HT2C receptor stimulated InsP3 production and caused $Ca^{2+}$ release from the sarcoplasmic reticulum. Selective activation of the Gs-coupled 5-HT4 receptor also stimulated cAMP production, and caused an increase in neuromodulin secretion. These findings demonstrate the ability of 5-HT receptor subtypes expressed in neurons to induce neuromodulin production. This leads to the activation of single or multiple G-proteins which regulate the $InsP3/Ca^{2+}/PLC-{\gamma}$ and adenyl cyclase / cAMP signaling pathways.

• Journal of Animal Science and Technology
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• v.46 no.1
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• pp.83-90
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• 2004

Processed meals, such as a ground meat and hamburger patty, are required to ensure that no pathogens remain in the final products. However, there was no rapid method available to verify that the recommended end-point cooking temperature(EPT) was reached. Thus, the objective of this study was to rapidly determine EPT of ground pork hams using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SOS-PAGE), based on the disappearance of sarcoplasmic proteins after cooking. Fresh pork hams were added two levels of salt(0, 2%) and fat(15, 25%) combinations, and stored in refrigerator overnight, and cooked to internal cooking temperatures of $64^{\circ}C$ to $74^{\circ}C$ with $2^{\circ}C$ increments. Cooked pork hams were measured cooking loss(CL, %), protein solubility(PS) and SOS-PAGE. CL(%) was reduced with the addition of 2% salt, as compared to the control, regardless of fat contents. It was also increased with increasing eooking temperature. Protein solubility was affected by the cooking temperature, resulting in reduced PS up to $64^{\circ}C$(P < 0.05), but remained constant higher than $68^{\circ}C$. In SOS-PAGE analysis, protein bands with the molecular weights of 36 and 66 kDa were affected by the addition of salt and fat combinations. regardless of treatments. These protein fractions were decreased gradually with increased cooking temperatures up to $68^{\circ}C$ ${\sim}$ $70^{\circ}C$ and might be good indicators for the determination of EPT in ground pork hams.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.190-196
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• 1983

This experiment was carried out to compare the change of muscle protein compositions, amino acid compositions of muscle protein, and free amino acid compositions by age of the Israeli strain of common carp, Cyprinus carpio nudus. Protein compositions of the muscle were: sarcoplasmic protein $25.8-27.2\%$, myofibrillar protein $62.3-56.2\%$, residual intracellular protein $9.6-13.2\%$ and stroma $2.3-2.9\%$. In between 1 year and 3 years, there were differences as follows; myofibrillar protein in 1 year was much than 3 years, and other proteins in 3 years were much than 1 year. By SDS-polyacrylamide gel electrophoresis, sarcoplasmic protein of the samples in 1 year an 3 years were composed of 11 subunits and 10 subunits, respectively. And appeared 210,000 dalton component in 1 year but did not appeared in 3 years. Myofibrillar protein was composed of 23 subunits in both 1 year and 3 years but the differences of subunits by age were not observed. No differences were observed by age in the composition of myofibrillar protein and residual intracellular protein. Amino acid composition of muscle protein in both 1 year and 3 years were no differences to each other, but the contents of glutamic acid, aspartic acid, lysine were higher than other amino acids. The amount of total free amino acid in 1 year was much than in 3 years.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.6
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• pp.539-547
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• 2019

Anoctamin 5 (ANO5)/TMEM16E belongs to a member of the ANO/TMEM16 family member of anion channels. However, it is a matter of debate whether ANO5 functions as a genuine plasma membrane chloride channel. It has been recognized that mutations in the ANO5 gene cause many skeletal muscle diseases such as limb girdle muscular dystrophy type 2L (LGMD2L) and Miyoshi muscular dystrophy type 3 (MMD3) in human. However, the molecular mechanisms of the skeletal myopathies caused by ANO5 defects are poorly understood. To understand the role of ANO5 in skeletal muscle development and function, we silenced the ANO5 gene in C2C12 myoblasts and evaluated whether it impairs myogenesis and myotube function. ANO5 knockdown (ANO5-KD) by shRNA resulted in clustered or aggregated nuclei at the body of myotubes without affecting differentiation or myotube formation. Nuclear positioning defect of ANO5-KD myotubes was accompanied with reduced expression of Kif5b protein, a kinesin-related motor protein that controls nuclear transport during myogenesis. ANO5-KD impaired depolarization-induced $[Ca2^{+}]_i$ transient and reduced sarcoplasmic reticulum (SR) $Ca^{2+}$ storage. ANO5-KD resulted in reduced protein expression of the dihydropyridine receptor (DHPR) and SR $Ca^{2+}-ATPase$ subtype 1. In addition, ANO5-KD compromised co-localization between DHPR and ryanodine receptor subtype 1. It is concluded that ANO5-KD causes nuclear positioning defect by reduction of Kif5b expression, and compromises $Ca^{2+}$ signaling by downregulating the expression of DHPR and SERCA proteins.

• Journal of Fisheries and Marine Sciences Education
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• v.3 no.2
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• pp.47-72
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• 1991

To obtain the principal data for useful treatment and processing of Korean geoduck (Panope japonica A. ADAMS) which inhabit mostly at Dong-Hae coastal area in Korea, changes of $NH_2$-N, TMAO, TMA, total creatinine, protein composition and fatty acid composition in raw and blanched geoduck muscle during storage at $-20^{\circ}C$ were investigated. In addition, its chemical composition variation in the whole year was elucidated. The moisture content in geoduck muscle meat was 78.1% to 82% in the whole year. Particularly, in July its moisture content was maximum as 82% and in September minimum as 78.1%. Crude protein was in the range of 12.3-16.4%, crude lipid the average was 1.5%, crude ash on the average was 1.4%. The abundant fatty acids in geoduck muscle oil were $C_{16}$ : 0, $C_{16}$ : 1, $C_{18}$ : 0, $C_{18}$ : 1, $C_{20}$ : 5, and $C_{22}$ : 6 acids. During storage at $-20^{\circ}C$, content of unsaturated fatty acid such as eicosapentaenoic acid (EPA, $C_{20}$ : 5) and docosahexaenoic acid (DHA, $C_{22}$ : 6)in raw geoduck muscle decreased somewhat and the raw geoduck was slightly oxidized. Trimethylamine (TMA), volatile basic nitrogen (VBN)and $NH_2$-N of raw muscle increased compared to blanched muscle. Trimethylamine oxide (TMAO) was slightly decreased during the storage period. The muscle protein was approximately composed of 37% sarcoplasmic, 29% myofibrillar, 22% alkali soluble, and 12% stroma protein. Among several proteins, myofibrillar protein content decreased mostly, while the alkali-soluble and stroma protein content increased slightly during storage at $-20^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.6
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• pp.550-556
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• 1985

In order to evaluate the marine mollusc muscle as foodstuff not only from the biochemical aspect but also from the view point of food science, we have analyzed the protein and amino acid compositions of the echiurid (Urechis unicinctus) and sea hare (Aplysia kurodai) muscle. The protein quality of the muscles was also investigated using in vitro methods based on in vitro digestibility, predicted digestibility, computed PER (C-PER) and discriminant computed PER(DC-PER). The remarkable feature of the protein compositions of the both muscles was that water soluble protein occupied a large amount of the muscle protein with fairly lower contents of the salt soluble protein. From the analysis of SDS-PAG electrophoresis, the sarcoplasmic proteins in the echiurid and the sea hare muscles were composed of 15 and 10 subunits, respectively. The free amino acid compositions of the total amino acids in the echiurid and sea hare muscle were characterized with $75\%$ of glycine and alanine, and with $78\%$ of taurine, respectively. The amino acid anaylsis of both muscle proteins showed that the echiurid muscle was rich in glycine, aspartic acid, glutamic acid, arginine and lysine, but was poor in cysteine, while the sea hare muscle was rich in glycine, glutamic acid, aspartic acid and arginine, but was negligible in cysteine and tryptophan. In the total amino acid profiles of the freeze dried muscles in echiurid and sea hare, there was not found a significant difference compared to the amino acid compositions of the muscle proteins. Predicting the protein quality of the echiurid and sea hare muscles using the in vitro method, it was apparently low compared to the muscle protein of fishes.

• Biomolecules & Therapeutics
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• v.26 no.6
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• pp.546-552
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• 2018

A comprehensive collection of proteins senses local changes in intracellular $Ca^{2+}$ concentrations ($[Ca^{2+}]_i$) and transduces these signals into responses to agonists. In the present study, we examined the effect of sphingosine-1-phosphate (S1P) on modulation of intracellular $Ca^{2+}$ concentrations in cat esophageal smooth muscle cells. To measure $[Ca^{2+}]_i$ levels in cat esophageal smooth muscle cells, we used a fluorescence microscopy with the Fura-2 loading method. S1P produced a concentration-dependent increase in $[Ca^{2+}]_i$ in the cells. Pretreatment with EGTA, an extracellular $Ca^{2+}$ chelator, decreased the S1P-induced increase in $[Ca^{2+}]_i$, and an L-type $Ca^{2+}$-channel blocker, nimodipine, decreased the effect of S1P. This indicates that $Ca^{2+}$ influx may be required for muscle contraction by S1P. When stimulated with thapsigargin, an intracellular calcium chelator, or 2-Aminoethoxydiphenyl borate (2-APB), an $InsP_3$ receptor blocker, the S1P-evoked increase in $[Ca^{2+}]_i$ was significantly decreased. Treatment with pertussis toxin (PTX), an inhibitor of $G_i$-protein, suppressed the increase in $[Ca^{2+}]_i$ evoked by S1P. These results suggest that the S1P-induced increase in $[Ca^{2+}]_i$ in cat esophageal smooth muscle cells occurs upon the activation of phospholipase C and subsequent release of $Ca^{2+}$ from the $InsP_3$-sensitive $Ca^{2+}$ pool in the sarcoplasmic reticulum. These results suggest that S1P utilized extracellular $Ca^{2+}$ via the L type $Ca^{2+}$ channel, which was dependent on activation of the $S1P_4$ receptor coupled to PTX-sensitive $G_i$ protein, via phospholipase C-mediated $Ca^{2+}$ release from the $InsP_3$-sensitive $Ca^{2+}$ pool in cat esophageal smooth muscle cells.

• The Korean Journal of Physiology
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• v.20 no.1
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• pp.31-41
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• 1986

The sufficient myoplasmic $Ca^{++}$ to react with the contractile proteins is necessary to induce contraction of a cardiac muscle. These $Ca^{++}$ for the production of muscle contraction are supplied from the three recognized $Ca^{++}$ sources; internal $Ca^{++}$ release via the sarcoplasmic reticulum(SR), $Ca^{++}$ influx through a gated Ca-channel in the membrane as a Isi, and $Ca^{++}$ transport by the mechanism of Na/ca exchange. However, it is still controversial which $Ca^{++}$ sources act as a main contributor for myoplasmic $Ca^{++}$, Therefore, this study was undertaken in order to examine the $Ca^{++}$ sources for the contraction of frog ventricle. There is evidence that the SR is sparse in frog ventricular fibers, and that T-tubules are absent. Isolated ventricular strips of frog, Rana nigromaculata, were used in this experiment. Isometric tension was recorded by force transducer, and membrane potentials of ventricular muscles were measured through the intracellular glass microelectrodes, which were filled with 3M KCI and had resistance of $30{\pm}50M{\Omega}$. All experiments were performed at room temperature in a tris·buffered Ringer solution which was aerated with 100% $O_2$. Isotonic high K, low Na solution was used to induce K-contracture, K-contracture appeared at the concentration of 20 to 30mM-KCI and was potentiated in parallel with the increase in KCI concentration. The contracture had two components: an initial rapid phasic and a subsequent slow tonic contractile responses. Membrane Potentials measured at normal Ringer solution(2.5mM KCI) was -90 to -100 mV, and decreased linearly as the KCI concentration increased; -55mV at 20mM.KCI, -45mV at 30 mM.KCI, -30 mY at 50 mM.KCI, and -12 mV at 100 mM.KCI. K-contracture was evoked firstly at the membrane potential of -45 mV. The contracture was potentiated by the increase of bathing extracellular $Ca^{++}$ concentration. However, in the absence of $Ca^{++}$ the contracture was almost not induced by 50 mM.KCI solution. Caffeine(20mM) in normal Ringer solution, which is known to release $Ca^{++}$ from SR without substantial effects on the $Ca^{++}$ fluxes across the surface membrane, did not affect membrane potential and also not initiate contracture, but the caffeine in 20 mM-KCI Ringer solution produced a contracture. Above results suggest that the main $Ca^{++}$ source for the K·contracture of frog ventricle is $Ca^{++}$ influx through the voltage-dependent Ca-channel, and that in the K-contracture at the concentration of 100 mM-KCI, the mechanism of Na/ca exchange also partly contributs, in addition to the $Ca^{++}$ influx.