• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.92-106
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• 2006

Purpose : To address the ability of Olibanum to induce cell death, we investigated the effect of olibanum on cell apoptosis. Twenty-four hours later, apoptosis occurred following olibanum exposure in a dose-dependent manner. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : The treatment of BAPTA-AM regulated olibanum-induced apoptosis in HeLa human cervical carcinoma cells. The 24 hr-earlier -thapsigargin-pretreated cell showed the resistance against olibanum-induced apoptosis and the Ru360-mitochondrial uniporter-inhibited olibanum-induced apoptosis, too. It means that olibanum leads to the accumulation of calcium and the resultant apoptosis in HeLa cells. Immunoblotting data also shows that the expression of GRP78, ER stress marker protein, was induced by the olibanum. Bcl-2, anti-apototic protein, was decreased and that the expression of Bax, pro-apoptotic protein, was increased by the addition of olibanum. Interestingly, the olibanum increased the activity of caspase-8 as well as calpain cysteine pretense in HeLa cervical carcinoma cells. Calpain inhibitor-calpastatin as well as caspase-8C/A expression abrogated olibanum-induced apoptosis in the carcinoma cells. The inhibition of caspase-8 regulated olibanum-induced calpain activation but the inhibition of calpain did not have any effect on the caspase-8 activation in HeLa human cervical carcinoma cells. Conclusion : We conclude that olibanum induces the accumulation of calcium and the resultant apoptosis in which caspase-8 and calpain are involved.

• The Korean Journal of Zoology
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• v.36 no.3
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• pp.347-352
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• 1993

N-ethylmaleimide at low concentrations is known to interact specifically with 2 sulfhydryl residues in myosin heavy chain. Calpain, a Ca$^2$+-dependent neutral protease isolated from chick skeletal muscle, was found to preferentially degrade the alkylated protein but much less significantly the unmodified protein. Exposure of myosin to KMnO$_4$, which is also known to interact with sulthydryl groups, also caused the rapid degradation of the myosin heavy chain. Furthermore, treatment of each agent with increasing concentrations results in a greater loss of the myosin ATPase activity, indicating that the modification occurred at the reactive site sulfhydryl residues. These results suggest that the covalent modification at the reactive site salfhydryl residues in the myosin heavy chain may mark the protein for degradation by intracellular proteases such as calpain.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.127-127
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• 1998

Specific inhibitors of a calcium activated neutral protease calpain could be used for the treatment of neurodegenerative diseases, cataract and muscular dystrophy diseases because of their therapeutic effects. In the course of screening for potential calpain inhibitors from microorganisms, a new analogue of chymostatins named calmostinol was isolated from the culture filtrate of an actinomycete. The MW was determined to be 596 [(M + H)$\^$+/] by FAB-MS in glycerol matrix. The structure was elucidated to be N-[((S)-1-carboxy-2-phenylethyl)-carbamoyl]-${\alpha}$-[2- iminohexahydro-4(S)-pyrimidyl]-L-glycyl- L-valyl-phenylalaninol, by the spectroscopic methods such as NMR and MS fragmentation studies. Calmostinol exhibited strong activity against calpain while not against a Ca$\^$2+/ -independent cysteine protease papain.

• Natural Product Sciences
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• v.18 no.2
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• pp.102-105
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• 2012

Marine organism extracts were prepared from 26 species of Korean indigenous marine organisms, including 25 species belonging in class Anthozoa of phylum Cnidaria and a species belonging to subphylum Urochordata of phylum Chordata, and screened their inhibitory effects against ${\mu}$-calpain. As a result, the thirteen extracts were found to be active in the criteria of $IC_{50}$ < 100 ${\mu}g/ml$. Among them, the MeOH extracts of Plexauroides praelonga and Alveopora japonica showed remarkable ${\mu}$-calpain inhibitory activity with $IC_{50}$ values of $4.62{\pm}0.22$ and $4.82{\pm}0.07{\mu}g/ml$, respectively. In addition, chemical investigation of A. japonica led to the isolation of an active compound, hexadecyl tetradecanoate, as a selective cathepsin B inhibitor ($IC_{50}=9.05{\pm}2.45{\mu}M$). This compound was isolated as constituent of A. japonica for the first time in the present study.

• Journal of Life Science
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• v.20 no.10
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• pp.1498-1504
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• 2010

A number of studies have shown that the calpain system is important in normal skeletal muscle growth. An increased rate of skeletal muscle growth can result from a decreased rate of muscle protein degradation, and this is associated with a decrease in activity of the calpain system, due principally to a large increase in calpastatin (CAST) activity. The CAST gene, mapped to BTA 7, is considered a candidate gene for beef tenderness and muscle growth. The present study used comparative sequencing of five novel polymorphisms located within exon 20 and 22 of the bovine CAST gene in Hanwoo: exon20- 109737G/A, 109749T/C, 109823T/C, exon22- 116151G/A, intron- 109926G/A. The association of the CAST SNPs with economic traits was studied. The 109926G/A showed a significant effect only on the longissimus muscle area (LMA, p<0.05) in Hanwoo. 109926G/A with the genotype GG had a significantly higher effect on LMA (75.35) than the genotype AA (69.6, p<0.05). Also, the 116151G/A showed a significant effect only on weight at 18 months (W18, p<0.05). 116151G/A with the genotype GG had a significantly higher effect on W18 (428.54) than the genotype AA (408.87, p<0.05).

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.67-67
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• 2003

Minocycline is known to protect neurons from microglia-mediated cell death in many experimental models of brain diseases including ischemic stroke, Huntingtons disease (HD), amyotrophic lateral sclerosis (ALS), traumatic brain injury, multiple sclerosis, and Parkinsons disease. When the activity of caspases was assessed using their fluorescent peptide substrates, activation of caspase-2, 3, 8, and 9 was evident within 2 8 hr following oxidative insult with 0.5 mM hydrogen peroxide in PC12 cells. Minocycline significantly attenuated activation of these caspases up to 18 hr, resulting a significant increase in the cell viability as assessed by MTT assay as well as trypan blue staining. However, cleavage of alpha-spectrin and a cdk5 activator p35, which are known to be substrates for calpain, remained unchanged in the presence of minocycline, suggesting that minocycline did not block caspase-3-independent cell death or necrosis. Moreover, co-treatment with minocycline and a calpain inhibitor calpeptin synergistically inhibited hydrogen peroxide-induced cell death. These data suggest that minocycline directly inhibited apoptosis, but not necrosis, after oxidative insult in PC12 cells.

• Proceedings of the Zoological Society Korea Conference
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• 2003.08a
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• pp.177.1-177
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• 2003

No Abstract, See Full Text

• The Journal of Korean Medicine
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• v.23 no.3
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• pp.145-163
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• 2002

Objectives: The purpose of this investigation was to evaluate the effects of Woohwangcheongsim-won and to study the mechanism for neuronal death protection in hypoxia with Embryonic day 20 (E20) cortical cells of a rat (Sprague Dawley). Methods: E20 cortical cells were dissociated in neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Woohwangcheongsim-won was added to the culture media for 24 hrs or 72 hrs. On 17 DIV, cells were given a hypoxic shock and further incubated in normoxia for another three days. On 20 DIV, Woohwangcheongsim-won's effects for neuronal death protection were evaluated by LDH assay, propidium iodide stain and phospho-H2AX immunostain and the mechanisms were studied by Bcl-2, Bak, Bax, caspase family, PKCα, ca1pain I. Results & Conclusions : 1. This study indicated that Woohwangcheongsim-won's effects for neuronal death protection in hypoxia were confirmed by LDH assay, propidium iodide stain and phospho-H2AX immunostain in culture method of Embryonic day 20(E20) cortical neuroblasts. 2. Woohwangcheongsim-won's mechanisms for neuronal death protection in hypoxia are to reduce the membrane damage fraction, to restrain DNA truncate, to restrain inflow of cytochrome c into cellularity caused by Bak diminution, to reduce the caspase cascade intiator caspase-8 and the effector caspase-3, to reduce the calpain I activity and to increase PKCand its activity in the membrane fraction. (J Korean Oriental Moo 2002;23(3):145～163)

• Food Science of Animal Resources
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• v.40 no.3
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• pp.415-425
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• 2020

It is believed that two main proteolytic systems are involved in the tenderization of meat: the cathepsins and the calpains. Many researchers consider the calpain system to be the major contributor to meat tenderness during post-mortem storage. However, the role and activity of cathepsins during post-mortem storage or low temperature meat processing is unclear, particularly for the tough meat cuts like brisket. Thus, the study was designed to investigate the effects of cold (refrigerated and frozen) storage and sous vide processing on the activities of cathepsin B, H, and L in beef brisket. There were no significant changes in pH and cathepsin H activity throughout the 18 d of storage at both temperatures. However, an increase in cathepsin B activity was observed during the first 4 d at both storage temperatures, but subsequently the activity remained unchanged. Cathepsins B and L were found to be more heat stable at sous vide temperatures (50℃ for 24 h, 55℃ for 5 h and at 60℃ and 70℃ for 1 h) compared to cathepsin H. Cathepsin B+L activity was found to increase after sous vide cooking at 50℃ for 1 h but decreased to about 47% relative to the uncooked control after 24 h of cooking. These results suggest that cathepsins B and L may contribute to the improved meat tenderness usually seen in sous vide cooked brisket meat.

• Food Science of Animal Resources
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• v.41 no.4
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• pp.589-607
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• 2021

Meat proteolytic systems play a crucial role in meat tenderisation. Understanding the effects of processing technologies and post-mortem storage conditions on these systems is important due to their crucial role in determining the quality characteristics of meat and meat products. It has recently been proposed that tenderisation occurs due to the synergistic action of numerous endogenous proteolytic systems. There is strong evidence suggesting the importance of μ-calpain during the initial post-mortem aging phase, while m-calpain may have a role during long-term aging. The caspase proteolytic system is also a candidate for cell degradation in the initial stages of conversion of muscle to meat. The role of cathepsins, which are found in the lysosomes, in post-mortem aging is controversial. Lysosomes need to be ruptured, through aging, or other forms of processing to release cathepsins into the cytosol for participation in proteolysis. A combination of optimum storage conditions along with suitable processing may accelerate protease activity within meat, which can potentially lead to improved meat tenderness. Processing technologies such as high pressure, ultrasound, and shockwave processing have been reported to disrupt muscle structure, which can facilitate proteolysis and potentially enhance the aging process. This paper reviews the recent literature on the impacts of processing technologies along with post-mortem storage conditions on the activities of endogenous proteases in meat. The information provided in the review may be helpful in selecting optimum post-mortem meat storage and processing conditions to achieve improved muscle tenderness within shorter aging and cooking times.