• Asian-Australasian Journal of Animal Sciences
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• v.19 no.5
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• pp.739-743
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• 2006

Postmortem proteolysis of breast (BM) and leg (LM) muscles from Taiwan colored chickens (TCC) and silkie bantams (SB) at $5^{\circ}C$ were compared. Myofibrils were prepared from BM and LM samples that were randomly taken from the carcasses of SB and TCC after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. pH of samples was determined, and degradation of myofibrillar proteins was analyzed by the SDS-PAGE and western blots. The results showed that pH was lower in BM than in LM samples from both avian strains. Appearance of 30 kDa components and disappearance of titin and nebulin were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data might suggest that postmortem proteolysis occurred more rapidly in BM than in LM from both TCC and SB.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.721-724
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• 2002

Postmortem Proteolysis of breast (BM) and leg (LM) muscles in Chiayi native chickens at $5^{\circ}C$ were compared. Myofibrils were purified from BM and LM samples that were randomly taken from carcasses after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. Fragmentation of myofibrils were determined, and degradation of myofibrillar proteins were analyzed by the SDS-PAGE and western blots. The results showed that myofibril fragmentation index (MFI) was significantly (p<0.05) higher in BM than in LM samples. Disappearance of titin and nebulin and appearance of the 30 kDa component were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data suggested that postmortem proteolysis occurred more rapidly in breast muscles in Chiayi native chickens.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.594-599
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• 2006

In our previous report, we indicated that not only BSA but also BGG played an important role in the allergenicity of beef. In this study, the effect of heat or high-pressure treatments to beef extract on the SDS-PAGE patterns was examined. The antigenicity of each treated samples was also investigated by Western blots assay with the sera of BGG-positive beef allergic patients. The BGG band and its antigenicity slightly disappeared but not generally in $100^{\circ}C$ group, indicating $100^{\circ}C$ treatment is not sufficient to totally eliminate the antigenicity of beef allergens. Compared with BGG band, BSA band significantly disappeared in SDS-PAGE with $100^{\circ}C$ treatment, indicating BSA is more heat- sensitive than BGG. When the beef extract was heated at $120^{\circ}C$, not only BSA but also BGG bands was largely disappeared in both SDS-PAGE and Western blots. High pressure (HP) treatment even at 600 MPa did not affect SDS-PAGE and Western blots pattern of BSA. On the contrary, BGG treated with HP showed visible changes in SDS-PAGE. 600 MPa treatment significantly reduced the antigencity. Interestingly, these behaviors of BGG were not found in the same experiments with pure BGG treated with HP. From these results, it was speculated that some kinds of proteolytic enzymes in beef extracts were involved in the BGG molecular degradation by HP treatment. The aging experiments of beef extracts treated with HP supported this hypothesis. Further studies are needed to clarify the function and working mechanism of enzymes associated with BGG degradation in beef extracts by HP treatment.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.131-146
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• 1998

To examine the effect of Shiquandabutang on the metastasis of cancer, the following experiments were made. Before the main experiments, the cytotoxicity was measured by putting Shiquandabutang sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. And western blotting was carried out to examine the changes of Fos, Jun, Ets, the transcription factors of MMP-2, MMP-9, and Erk, JNK on signal transduction pathway to AP-1. Third, in vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the results of the above the following conclusions were obtained. 1. The experimental result about cytotoxicity of Shiquandabutang against HT1080 was as below. The stained cell count after being treated by Shiquandabutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Shiquandabutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Shiquandabutang sample $400{\mu}g/ml$, MMP-2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disappeared. In Shiquandabutang sample $800{\mu}g/ml$, both bands of MMP-2 and MMP-9 disappeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were as below. In Shiquandabutang sample $200{\mu}g/ml$, Ets was reduced, and Fos were increased. 4. The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Shiquandabutang-treated group was less than that of +TPA control group. From the above results, it was concluded that Shiquandabutang might control the appearing and acting of collagenase not by the MMP-2, -9 promoter but by other way.

• Herbal Formula Science
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• v.25 no.4
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• pp.537-551
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• 2017

Objectives : Jinnoe-san (JNS) is a novel herbal formula consisting of five oriental medicinal herbs including Polygalae Radix, Prunellae Spica, Perillae Herba, Betulae Cortex, and Lonicerae Flos. In this study, we investigated the effects and molecular mechanism of JNS on Parkinson's disease in vitro model. Methods : The effects of JNS on 1-methyl-4-phenylpyridinium ($MPP^+$)-induced cell death in SH-SY5Y cells were evaluated with a cell viability assay, flow cytometry, and western blots analysis. The effects of JNS on lipopolysaccharide (LPS)-stimulated BV2 microglia were determined with a nitric oxide (NO) assay, enzyme linked immunosorbent assays, and western blots analysis. Result : $MPP^+$-induced cell death in SH-SY5Y cells was significantly reduced by JNS pre-treatment in a dose-dependent manner. JNS inhibited the production of reactive oxygen species, mitochondria dysfunction, and apoptosis induced by $MPP^+$ in SH-SY5Y cells. Furthermore, JNS significantly activated Akt and ERK in SH-SY5Y cells and the ability of JNS to prevent mitochondria dysfunction by $MPP^+$ was antagonized by pre-treatment of LY294002 and PD98059, an Akt and ERK inhibitor, respectively. In addition, JNS inhibited LPS-induced NO and $PGE_2$ production as well as iNOS expression and secretion of TNF-${\alpha}$, pro-inflammatory cytokines without affecting the cell viability. JNS also suppressed LPS-induced ERK activation. Conclusions : These results demonstrate that JNS has a protective effect on the dopaminergic neurons against $MPP^+$-induced neurotoxicity and anti-inflammatory effect on the LPS-stimulated microglia. These findings provide evidences for JNS to be considered as a new prescription for treating Parkinson's disease.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.222.1-222.1
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• 2003

The signal transduction pathway leading to the activation of the p70s6k plays an important role in the progression of cells from G0/G1 to S phase of the cell cycle but remains incompletely characterized. We investigated the role of the Rho family G protein Rac1 in H2O2-mediated p70s6k activation. Transient expression of a dominant negative mutants of the small GTP-binding proteins Rac1 (Rac1N17) and Cdc42(Cdc42N17) showed reduced levels of slower migration on Western blots of one-dimensional SDS-PAGE in p70s6k and ERK1/2 by PDFG stimulation. (omitted)

• Journal of Gastric Cancer
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• v.3 no.1
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• pp.44-49
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• 2003

Purpose: The aim of this study was to investigate the impact of survivin expression and the decrease or loss of KAI-1 on the clinical stage and the survival rate in gastric adenocarcinomas. Materials and Methods: Expressions of survivin and KAI-1 were immunohistochemically determined in 40 cases of gastric adenocarcinomas. The survivin and KAI-1 expressions were also analyzed by using western blots in 14 cases among them. Results: Resected gastric cancer specimens from 40 patients (intestinal type: 15 cases and diffuse type: 25 cases) were evaluated immunohistochemically. Survivin protein expressions were significantly higher in diffuse types (P=0.03) and in advanced clinical stages (UICC TNM II and III, P=0.02). In contrast, a decrease or loss of KAI-1 expression had no statistically significant correlation with the Lauren classification or the clinical stage. Survivin protein positivity was associated with an unfavorable prognosis. Decrease or loss of KAI-1 was associated with a shorter disease free survivial rate (P < 0.01). The western blot data (n=14) indicated that neither survivin protein over-expression nor KAI-1 down-expression had an significant correlation with the Lauren classification or the clinical stage. Conclusion: In gastric carcinomas, survivin over-expression and decrease or loss of KAI-1 were associated with unfavorable prognosis, being independent prognostic factors along with the clinical stage and the disease free survival rate.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.79-89
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• 2004

Saussurea lappa and Taraxacum mongolicum have been used for herbal medicinal treatments against cancers in East Asia. We performed this study to understand the molecular basis underlying the anti-tumor effects of two herbs and analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules by using an AGS gastric cancer cell line. The treatments of Saussurea lappa dramatically reduced cell viabilities in a dose and time-dependent manner, but Taraxacum mongolicum did not. FACS analysis and Annexin V staining assay also showed that Saussurea lappa induces apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Saussurea lappa increased expression of the p53 and its downstream effector p21/sup Waf1/, and that the both increased expression of apoptosis related Bax and cleavage of active caspase-3 protein. We also confirmed the translocation of Bax to mitochondria Collectively, our data demonstrate that Saussurea lappa induces growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

• Journal of Society of Preventive Korean Medicine
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• v.10 no.2
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• pp.51-62
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• 2006

Psoriasis is a chronic skin disease characterized by angiogenesis. It has been reported that growth factor as vascular endothelial growth factor(VEGF) and insulin like growth factor(IGF) II are overexpressed in psoriatic epidermis. To investigate the inhibitory effects of IGF-II induced VEGF and HIF-1${\alpha}$ expression by RR extracts, we performed MTS assay, western blots using HaCaT cells. RR extracts significantly reduced IGF-II induced HIF 1${\alpha}$ protein level via MAPK pathway in HaCaT cells. Also, RR extracts inhibited IGF-II induced VEGF mRNA and protein expression levels in the HaCaT keratinocytes. These results suggest that inhibition of HIF-1${\alpha}$ and VEGF expressions by RR extracts contributes to the anti angiogenic effects.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.418-422
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• 2015

In the course of screening for novel cell cycle inhibitors and apoptotic inducers, CR389, elucidated as 5-(1H-benzoimidazol-2-yl)-1H-pyridin-2-one, was generated as a new hit compound. Flow cytometric analysis and western blots of PA-1 cells treated with $60{\mu}M$ CR389 revealed an appreciable cell cycle arrest at the G2/M phase through direct inhibition of the CDK1 complex. In addition, activation of p53 via phosphorylation at Ser15 and subsequent up-regulation of p21^CIP1 showed that CR389 also induces p53-dependent-p21^CIP1-mediated cell cycle arrest. Furthermore, apoptotic induction in $60{\mu}M$ CR389-treated PA-1 cells is associated with the release of cytochrome c from mitochondria through up-regulation of the proapoptotic Bax protein, which results in the activation of procaspase-9 and -3, and the cleavage of poly(ADP-ribose) polymerase (PARP). Accordingly, CR389 seems to have multiple mechanisms of antiproliferative activity through p53-mediated pathways against human ovarian cancer cells. Therefore, we conclude that CR389 is a candidate therapeutic agent for the treatment of human ovarian cancer via the activation of p53.