• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1653-1658
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• 2002

The objectives of this study were to examine calpain activity and meat tenderness by three different feeding patterns in Korean native cattle (KNC). Total forty-five animals were assigned each fifteen in long term restriction feeding (LTFR), long-term restriction feeding and hormone treatment (LTFR-tH), and short term non-restriction feeding (STFNR), respectively. Concentrate was restricted based on body weight in exp 1 and 2. However, it was fed ad libitum in exp. 3. Hormonal implantation was made with $M-PO^{TM}$ for bulls and with $F-TO^{TM}$ for heifers at 18, 20, 22 months of age in exp. 2. Animals were purchased (3-5 month old) from local cattle market and managed in two local farms and university research unit at three different years. Animals were slaughtered at 24 months for long-term trial and at 18 month for short term-trial. Loin and tender loin muscle was used for calpain activity and meat quality. Calpain proteolytic system was not changed by treatment. However, calpastatin activity was low in short-term trial. The calpain and calpastatin activity is reciprocal relationship, therefore, the high calpain activity may effect on quality grade. The shear force value was decreased as the processing of aging after postmortem. On the other hand, the cooking loss was significantly higher in short-term than in long-term trial, and then gradually decreased by the aging. Hormone implants to increase meat yield influenced to calpastatin activity more powerfully than calpain activity to meat tenderness. In meat color-a*, there was not significant difference in loin. Meat color-b* was decreased as postmortem aging time increased in tenderloin. Western blots were done to learn whether these proteins are degraded during postmortem storage and whether this degradation temporally parallels the decrease of shear force value. Vinculin was detected at 0 day and 1 day and degraded after 3 day. In conclusion, Calpain activity was affected slightly on meat tenderness. But meat tenderness was influenced by calpastatin, more effectively.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1747-1753
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• 2002

The objectives of this study were to examine the effect of early short-term temperature conditioning on metabolic rate and calpain/calpastatin system and to compare variations in metabolic rate and calpain/calpastatin system between Hanwoo and Holstein beef. Longissimus thoracis et lumborum of the right carcass from 3 Hanwoo and 3 Holstein bulls were removed within 30 min of exsanguinations, cut into three pieces, and then temperature conditioned until 3 h postmortem (PM) at 2, 16, and $30^{\circ}C$, respectively. Rigor values (R-values; $R_248$, $R_250$, and $R_258$), pH, muscle temperature, glycogen content, $\mu$- and m-calpain activities, and calpastatin acitivity were measured at 1, 3, 9, and 24 h PM, respectively. Hanwoo beef had higher muscle temperature, faster metabolic rate at early PM stage in R-values, and lower $\mu$-calpain activity than Holstein beef (p<0.05). The $30^{\circ}C$ treatment maintained muscle temperature of $30^{\circ}C$ until 3 h PM and resulted in faster pH decline at 3 and 9 h PM (p<0.05) than other treatments. The $16^{\circ}C$ had higher (p<0.05) muscle temperature at 3 h PM than the $2^{\circ}C$, but no difference in all other traits was observed between the $2^{\circ}C$ and the $16^{\circ}C$. Early shortterm temperature treatment used in this study was not sufficient to effectively activate calpain/calpastatin system. Correlations among all traits except m-calpain and muscle temperature were generally high (r>0.60; p<0.001). Among R-values, $R_258$ had higher correlations with other metabolic traits than those of $R_248$ and $R_250$. These data suggest that early PM metabolic rate, $\mu$-calpain activity, and calpastatin activity may be closely related to each other. Variations in metabolic rate and $\mu$-calpain activity at early PM stage between Hanwoo and Holstein beef may imply variations in meat quality between both breeds.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.5
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• pp.753-762
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• 2020

Objective: The aim of the present study was to determine the effect of supplementing pasture-finished steers with corn silage on the expression level of the calpain system proteins and beef tenderization. Methods: Thirty Braford steers grazing on summer pasture were used for the study. For 120 days fifteen animals were supplemented with corn silage at 1% of body weight per head per day (Suppl) whereas the remaining 15 steers only received pasture (Contr). Carcass and meat traits were evaluated and compared between groups. Gene expression and activities of proteases (calpain 1 and calpain 2) and inhibitor (calpastatin) were measured using real-time polymerase chain reaction and casein zymography. Results: Carcass and meat traits were significantly different between feeding systems. Supplemented steers showed higher hot carcass weight (p<0.01), fat content (p = 0.02), and Warner-Bratzler shear force (p = 0.03). Furthermore, the control group showed higher protease:inhibitor ratios, at mRNA (p = 0.01) and protein levels (p<0.10). Warner-Bratzler shear force and mRNA calpains:calpastatin ratio were associated in both feeding systems (p<0.01). Conclusion: Based on the results obtained in the study, beef tenderness differences among finishing strategies could be modulated through differential expression of the calpain system proteins.

• Food Science of Animal Resources
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• v.38 no.3
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• pp.515-529
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• 2018

The present study was designed to investigate the effects of calpain system on the formation of volatile flavor compounds in Hanwoo beef. In the first experiment (exp.1), Longissimus lumborum (LL) muscle samples were injected with solutions containing 50 mM $CaCl_2$ or 50 mM $ZnCl_2$ and 154 mM NaCl respectively, and aged for 7 d at $4^{\circ}C$. In the second experiment (exp.2), the ground LL muscle was incubated with the aforementioned solutions containing cathepsin inhibitor. The injection with $CaCl_2$ solution greatly elevated the calpain activity and concomitantly, significantly decreased the Warner-Bratzler shear force (p<0.05). The pH, meat color and cooking loss did not differ (p>0.05) between the treatment groups. A total of 51 volatile compounds were identified using the solid phase microextraction with gas chromatography (SPME-GC). Results on volatile analyses from the both experiments showed that the injection with calcium ions led to significant increase (p<0.05) concentrations of pyrazines and sulfuric compounds. These results coincide with a higher rate of protein degradation due to the $CaCl_2$ injection as compared to the control group. Significantly (p<0.05) higher levels of lipid oxidation derived-aldehydes were found in the samples with $ZnCl_2$. The exp.1 showed that cathepsin inhibitors had no effect on the formation of volatile flavor components after 7 d of aging. These results imply that the proteolytic activity of the calpain system is associated with generation of volatile compounds of chiller-aged beef, while the role of cathepsins is likely very limited.

• 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.

• Food Science of Animal Resources
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• v.25 no.3
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• pp.310-315
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• 2005

The changes in pH, temperature, R-values and ${\mu}-calpain$ and its inhibitor activity of M. longissimus from Hanwoo steer were investigated at 1, 3, 9 and 24h postmortem. The pH and temperature of M. longissimus were significantly (p<0.05) decreased during 24h postmortem time, and were 6.50 and $31.99^{\circ}C$, respectively, at 3h postmortem. $R_{248}\;and\;R_{250}$ were increased, but $R_{258}$ was decreased after 9h postmortem time (p<0.05). Calpain I and calpastatin activity were decreased after 3h and 9h postmortem time, respectively (p<0.05). pH and temperature showed high positive correlations with $R_{258}$ (r=0.967 and r=0.970, respectively), calpain I (r =0.956 and r=0.954, respectively) and calpastatin (r=0.978 and r=0.986, respectively) but had high negative correlations with $R_{248}$ (r=-0.982 and r=-0.973, respectively) and $R_{248}$ (r=-0.983 and r=-0.976, respectively). from these results, the change of postmortem metabolism of M. longissimus from Hanwoo steer likely occurred after 9h postmortem time. However, the further study on the establishment of metabolism from Hanwoo between postmortem 3h and 9h are necessary to produce Hanwoo beef with high acceptance in meat quality.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.3
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• pp.443-454
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• 2013

Tenderness is the most important meat quality trait, which is determined by intracellular environment and extracellular matrix. Particularly, specific protein degradation and protein modification can disrupt the architecture and integrity of muscle cells so that improves the meat tenderness. Endogenous proteolytic systems are responsible for modifying proteinases as well as the meat tenderization. Abundant evidence has testified that calpains (CAPNs) including calpain I (CAPN1) and calpastatin (CAST) have the closest relationship with tenderness in livestock. They are involved in a wide range of physiological processes including muscle growth and differentiation, pathological conditions and post-mortem meat aging. Whereas, Calpain3 (CAPN3) has been established as an important activating enzyme specifically expressed in livestock's skeletal muscle, but its role in domestic animals meat tenderization remains controversial. In this review, we summarize the role of CAPN1, calpain II (CAPN2) and CAST in post-mortem meat tenderization, and analyse the relationship between CAPN3 and tenderness in domestic animals. Besides, the possible mechanism affecting post-mortem meat aging and improving meat tenderization, and current possible causes responsible for divergence (whether CAPN3 contributes to animal meat tenderization or not) are inferred. Only the possible mechanism of CAPN3 in meat tenderization has been confirmed, while its exact role still needs to be studied further.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.77-91
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• 2006

Purpose : To address the ability of Dohaekseungkitang (DST: a commonly used herb formulation in Korea, Japan and China to have anti-cancer effect on cervical carcinoma), we investigated the effects of DST on programmed cell death (apoptosis) in HeLa human cervical carcinoma cells. Methods : We cultured 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 : After the treatment of DST for 48 hours, apoptosis occurred in a dose-dependent manner. In this study, we have shown that DST induces calpain and the associated caspase-8 and -9 activations. Apoptosis was prevented by pre-incubation of the cells with the calcium cHeLator-BAPTA-AM, calcium channel blocker-Nif edipine or Ryonidine agonist-Ryonidine peptide, implicating calcium in the apoptotic process. Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, especially in calcium-related apoptosis. However this study showed 1hat either calpain inhibitor-calpastin or caspase-3 inhibitor-DEVD- did not blocked the herb formulation-induced apoptosis in HeLa human cervical carcinoma cells. D ST initiates a cell death pathway that is partially dependent of caspases. DST-induced apoptosis requires caspase-independent mechanism. Conclusion : We conclude that DST-induced calpain activation triggers the intrinsic apoptotic pathway in which caspase-independent mechanism is also involved.

• Journal of Pharmacopuncture
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• v.13 no.1
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• pp.15-35
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• 2010

연구목적 : 이 연구는 봉약침의 봉독이 NF-${\kappa}B$ 활성억제와 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 발현을 통하여 세포자멸사를 유도하고, 전립선 암세포를 이식한 쥐에서의 세포자멸사 유도 효과를 확인함으로써, 봉약침의 봉독이 생체 내에서도 세포자멸사를 유도하여 전립선암에 효과를 나타냄을 확인하고자 하였다. 실험방법 : 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절 단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였다. 결과 : 1. DAPI, TUNEL staining assay 결과 봉독 및 melittin을 처리한 LNCaP 세포 모두에서 세포자멸사 유도율이 유의한 증가를 나타내었다. 2. LNCaP 세포에 봉독이나 melittin을 처리한 결과, 안드로겐 수용체 조절 단백질 중 p-Akt, COX-2, calpain은 봉독과 melittin 모두에서 유의한 감소를 나타내었고, Akt는 melittin에서 유의한 감소를 나타냈으며, 봉독에서 증가하는 경향을 보였고, MMP-9은 증가하였다. 3. 생체 내에서의 봉독의 항암효과를 확인하기 위해 전립선암세포가 이식된 쥐에 봉독을 처리한 후 암세포의 부피와 무게, 쥐의 체중을 측정한 결과, 봉독을 처리한 군에서 암 세포 부피비율 및 무게는 감소하였고, 쥐의 체중은 증가하였다. 4. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, NF-${\kappa}B$ 활성에서 유의한 감소를 나타내었다. 5. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, 세포자멸사 조절 단백질 중 Bax/Bcl-2, p53, caspase-3, caspase-9, calpain은 유의한 증가를, COX-2는 유의한 감소를 나타냈으며, MMP-9는 증가를 나타내었다. 결론 : 이상의 결과는 봉독이 시험관 내에서 뿐만 아니라 생체 내에서도 NF-${\kappa}B$의 활성을 억제하고 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 조절을 통하여 인간 전립선암 세포주인 LNCaP의 세포자멸사를 유도함으로써 전립선암 세포 증식억제 효과 및 호르몬 비의존적인 전립선암으로의 전이를 지연시키는 경향이 있을 것으로 사료되고, 봉독이 전립선암의 예방과 치료에 효과적으로 활용될 수 있을 것으로 기대된다.

• Korean Journal of Agricultural Science
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• v.40 no.2
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• pp.147-153
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• 2013

The aim of study was to investigate genetic diversity for the calpain/calpastatin gene in three Hanwoo breeds [(Brown (n=62), Brindle (n=81) and Jeju Black (n=30)]. Random samples from three breeds of Hanwoo were selected and genotyped for the 7 SNPs of calpain/calpastatin using TaqMan method. Allele frequencies were investigated for CAPN1/CAST gene. Allele frequency of CAST2 SNP was 0.75, 0.59 and 0.22 for Brown, Brindle and Jeju black, respectively. The CAST3 revealed allele frequency of 0.59 and 0.57 in Brown and Jeju Black, while it showed very low allele frequency (0.07) in Brindle. In particular, favorable allele (G allele) for the CAPN1-2 SNP which was shown a strong association with tenderness in Taurine and Indicine cattle revealed 16% and 17% higher allele frequency in Brown Hanwoo (0.82) comparing Brindle (0.66) and Jeju Black Hanwoo (0.65). AMOVA demonstrated that among population variance occupied only 10% of total variance and among individual variance was 0%, while within individual variance was 90% of total variance. This result showed that population effect contributed very small portion of genetic to these three Hanwoo breeds, while within individual variance contributed large portion of genetic diversity within these Hanwoo breeds. In conclusion, three Hanwoo breeds (Brown, Brindle and Jeju black) showed a genetically homogeneous based on the 7 SNPs of CAPN1/CAST gene and it came from same ancestor to form modern Hanwoo breed.