• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.319-324
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• 2010

Phorbol ester extraction was carried out from Jatropha curcas seed cake, a by-product from the biodiesel fuel industry. Four repeated extractions from 5 g of J. curcas seed cake using 15 ml of 90% (v/v) ethanol and a shaking speed of 150 rpm gave the highest yield of phorbol esters. The ethanolic extract of J. curcas seed cake showed antifungal activities against important fungal phytopathogens: Fusarium oxysporum, Pythium aphanidermatum, Lasiodiplodia theobromae, Curvularia lunata, Fusarium semitectum, Colletotrichum capsid, and Colletotrichum gloeosporioides. The extract contained phorbol esters mainly responsible for antifungal activities. The extract could therefore be used as an antifungal agent for agricultural applications.

• Korean Journal of Pharmacognosy
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• v.35 no.1 s.136
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• pp.62-79
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• 2004

Cell-scattering is a phenotypic change easily observed in most epithelial cells treated with Hepatocyte Growth Factor /scatter Factor (HGF/SF) or phorbol esters (PKC-activators). Recent studies have shown the possibilities to use as therapeutic materials of HGF/SF or non tumor promoting phorbol esters for liver disease, cancer and AIDS. In this study, we tested a cell-scattering activity of 534 methanol extracts from plants inhabiting in Korean peninsula using the phenotype-based assay system. Nine Active extracts were detected : Daphne genkwa, Daphne kiusiana, and Aleurites fordii showed high activity (+++), Euphorbia sieboldiana and Rhodotypos scandens showed medium activity (++), Sambucus sieboldiana var. pendula, Catalpa bignonioides, Sambucus sieboldiana and Lycoris squamigera showed low activity (+). Furthermore, the effects of these active materials in the culture cells were investigated with biochemical studies.

• BMB Reports
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• v.38 no.6
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• pp.639-645
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• 2005

Protein kinase C (PKC) isozymes, a family of serine-threonine kinases, are important regulators of cell proliferation and malignant transformation. Phorbol esters, the prototype PKC activators, cause PKC translocation to the plasma membrane in prostate cancer cells, and trigger an apoptotic response. Studies in recent years have determined that each member of the PKC family exerts different effects on apoptotic or survival pathways. $PKC{\delta}$, one of the novel PKCs, is a key player of the apoptotic response via the activation of the p38 MAPK pathway. Studies using RNAi revealed that depletion of $PKC{\delta}$ totally abolishes the apoptotic effect of the phorbol ester PMA. Activation of the classical $PKC{\alpha}$ promotes the dephosphorylation and inactivation of the survival kinase Akt. Studies have assigned a pro-survival role to $PKC{\varepsilon}$, but the function of this PKC isozyme remains controversial. Recently, it has been determined that the PKC apoptotic effect in androgen-dependent prostate cancer cells is mediated by the autocrine secretion of death factors. $PKC{\delta}$ stimulates the release of $TNF{\alpha}$ from the plasma membrane, and blockade of $TNF{\alpha}$ secretion or $TNF{\alpha}$ receptors abrogates the apoptotic response of PMA. Molecular analysis indicates the requirement of the extrinsic apoptotic cascade via the activation of death receptors and caspase-8. Dissecting the pathways downstream of PKC isozymes represents a major challenge to understanding the molecular basis of phorbol ester-induced apoptosis.

• The korean journal of orthodontics
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• v.32 no.3 s.92
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• pp.227-234
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• 2002

In order to investigate the action mechanism of protein kinase C on $K^+$ channel in osteoblastic cell, effects of phorbol 12, 13-dibutyrate on human osteoblast-like cells (G292) were studied by patch clamp technique with cell-attacked configuration. 111 this experiment, 45pS ion channel was dominant in G292 cell line according to their approximate conductances in symmetrical 140mM KCl saline at holding potential of 60mV. In torrent-voltage relationship, reversal potential was 5.5mV at the condition of potassium enriched saline in the pipette and -27 mV at the condition of standard extracellular saline In the pipette. Phorbol 12, 13-dibutyrate 10nM increased the open probability of 45pS channel and staurosporine, an inhibitor of protein kinase C, suppressed this effect. Phorbol 12,13-dibutyrate moved the reversal potential of 45pS channel to more negative potential and increased the single channel current at the same membrame potential. In order to check the activation of protein kinase C in G292 cell by phorbol 12,13-dibutyrate, western blot of protein kinase C was performed. Phorbol 12,13-dibutyrate $0.1{\mu}M$ translocated protein kinase C from cellular compartment to membrane compartment of the cell. These findings suggest that phorbol 12,13-dibutyrate, one of phorbol esters, activate 45pS channel In G292 cell and affect cell membrane potential, that regulate cellular function.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.6
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• pp.888-898
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• 2018

Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

• Korean Journal of Biological Psychiatry
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• v.2 no.2
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• pp.252-256
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• 1995

Corticotropin-releasing factor(CRF)를 분비하는 배양세포주는 CRF의 세포내 조절을 연구하는데 있어 훌륭한 체계가 된다. BE(2)-M17 및 BE(2)-C 세포주는 CRF를 생산하고 분비하며 forskolin 처치에 반응하는 것으로 알려져 있다. 저자들은 이들 세포주에 phorbol 에스테르, 즉 phorbol 12-myristate 13-acetate(TPA) 0.8, 4, 10, 20 nM, 및 nitric oxide(NO)의 신호전달 경로의 자극제(1-${\mu}M$ nitroprusside)와 차단제($1{\mu}M$ nitroprusside+$300{\mu}M$ $N^G$-methyl-D-arginine), 그리고 interleukin-$1{\alpha}$ (IL $1{\alpha}$ ; 4, 20, 100, 500 pM)를 처치하여 CRF를 의미있게 증가시켰으나 세포외 분비는 C 세포에서만 변화되었다. NOS계의 자극제와 차단제는 C 세포의 forskolin 효과를 의미있게 변화시켰다. IL$1{\alpha}$는 두 세포주에 대한 영향이 없었다. 상기 제제들에 대한 이들 세포의 반응이 중추신경계 CRF 신경들과 관련하여 논의되었다.

• Bulletin of the Korean Chemical Society
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• v.30 no.9
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• pp.1981-1984
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• 2009

Subcellular localization of protein kinase often plays an important role in determining its activity and specificity. Protein kinase C (PKC), a family of multi-gene protein kinases has long been known to be translocated to the particular cellular compartments in response to DAG or its analog phorbol esters. We used C-terminal green fluorescent protein (GFP) fusion proteins of PKC isoforms to visualize the subcellular distribution of individual PKC isoforms. Intracellular localization of PKC-GFP proteins was monitored by fluorescence microscopy after transient transfection of PKC-GFP expression vectors in the HeLa cells. In unstimulated HeLa cells, all PKC isoforms were found to be distributed throughout the cytoplasm with a few exceptions. PKC$\theta$ was mostly localized to the Golgi, and PKC$\gamma$, PKC$\delta$ and PKC$\eta$ showed cytoplasmic distribution with Golgi localization. DAG analog TPA induced translocation of PKC-GFP to the plasma membrane. PKC$\alpha$, PKC$\eta$ and PKC$\theta$ were also localized to the Golgi in response to TPA. Only PKC$\delta$ was found to be associated with the nuclear membrane after transient TPA treatment. These results suggest that specific PKC isoforms are translocated to different intracellular sites and exhibit distinct biological effects.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.59-65
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• 1993

We isolated Actinomycetes strain GMT 1155 and purified the active compound, MT 1155, on the morphological reversion of ts/NRK cell from the isolate. MT 1155 was identified as toyocamycin having antifungal and antitumor activities from physico-chemical properties and UV, IR, $^1H$-NMR, $^13C$-NMR and mass spectrum. MT 1155 showed the morphologically reversional activity on ts/NRK cell and the cytotoxicity on CTLL cell at the final concentrations of 1.7 JlM and 0.2 11M, respectively and its $IC_{50}$ value on protein kinase A enzyme was 2.3 $\mu$M. Also it had strong antifungal activity against several pathogenic fungi but not antibacterial activity. And it did not inhibit both protein kinase C activity and the bleb-formation of K562 cell induced by phorbol esters.

• The Korean Journal of Zoology
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• v.36 no.4
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• pp.439-451
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• 1993

Protein kinase C isozymes in a human prostate adenocarcinoma PC-3 cell line were characterized. Immunoreactive bands and immunocytochemical stains were obsenred in PC-3 cells with antibodies raised against protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$, and ζ types, respectively. Protein kinase C ${\alpha}$ corresponded to a immunoreactive band at a molecular weight of 80,000-dalton, whereas molecular weights of other immunoreactive isozvmes of protein kinase C were detected at 68,000-dalton. Protein kinHse C $\delta$ and ζ antibodies detected additional bands at 55,000-dalton and 80,000-dalton, respectively Immunocvtochemical study confirmed the results of the immunoblotting experiments qualitatively: all six protein kinase C isozymes were detected in the cytoplasm of PC-3 cells. Translocation of protein kinase C in PC-3 cells were also examined with phorbol 12-myristate 13-acetate (PMA), bryostatin 2, diolein, and 1-oleoyl-2-acetyl glycerol (OAG). Differential reactions of protein kinase C isozvmes to these activators were obsenred. When PC-3 cells were treated with 10mM bryostatin 2, protein kinase C isozyme u was translocated into the nucleus, whereas s type was translocated into the plasma membrane and the nucleus. Protein kinase C ${\alpha}$ and ζ types were translocated into the nucleus following the treatment with 101M diolein, whereas protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, and $\varepsilon$ types were translocated into the nucleus by the treatment with 10mM OAG. Protein kinase C ${\alpha}$ and $\varepsilon$ types were translocated into the nucleus in the presence of 100nM PMA. Protein kinase C $\delta$ type was translocated to the nuclear membrane by these activators, however, only PMA-induced translocation was inhibited by protein kinase C inhibitor, 1-(5-isoquinolinesulfonyll-2-methvlpiperazine dihvdrochloride (H7) . H7 inhibited translocation of protein kinase C ${\alpha}$ type induced by PMA, ${\beta}$ type by OAG and s type by PMA and OAG, whereas it did not affect translocations induced by bryostatin and diolein, respectively. These results suggest that there exist six isoformes of protein kinase C (${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$ and ζ types) in PC-3 cells and that each of these isozvmes distinctivelv reacts to bryostatin, diolein, OAG and PMA, in part due to an altered molecular size and conceivably discrete binding site(s).

• Tuberculosis and Respiratory Diseases
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• v.42 no.6
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• pp.862-870
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• 1995

Background: Oxygen free radicals have generally been considered as cytotoxic agents. On the other hand, recent results suggest that small nontoxic amounts of these radicals may act a role in intracellular signal transduction pathway and many efforts to reveal the role of these radicals as secondary messengers have been made. It is evident that the oxygen radicals are released by various cell types in response to extracellular stimuli including LPS, TNF, IL-1 and phorbol esters, all of which translocate the transcription factor $NF{\kappa}B$ from cytoplasm to nucleus by releasing an inhibitory protein subunit, $I{\kappa}B$. Activation of $NF{\kappa}B$ is mimicked by exposure to mild oxidant stress, and inhibited by agents that remove oxygen radicals. It means the cytoplasmic form of the inducible tanscription factor $NF{\kappa}B$ might provide a physiologically important target for oxygen radicals. At the same time, it is well known that LPS induces the release of oxygen radicals in neutrophil with the activation of $NF{\kappa}B$. From above facts, we can assume the expression of IL-8 and IL-$1{\beta}$ gene by LPS stimulation may occur through the activation of $NF{\kappa}B$, which is mediated through the release of $I{\kappa}B$ by increasing amounts of oxygen radicals. But definitive evidence is lacking about the role of oxygen free radicals in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. We conducted a study to determine whether oxygen radicals act a role in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. Method: Human peripheral blood monocytes were isolated from healthy volunteers. Time and dose relationship of $H_2O_2$-induced IL-8 and IL-$1{\beta}$ mRNA expression was observed by Northern blot analysis. To evaluate the role of oxygen radicals in the expression of IL-8 and IL-$1{\beta}$ mRNA by LPS stimulation, pretreatment of various antioxiants including PDTC, TMTU, NAC, ME, Desferrioxamine were done and Northern blot analysis for IL-8 and IL-$1{\beta}$ mRNA was performed. Results: In PBMC, dose and time dependent expression of IL-8 and IL-$1{\beta}$ mRNA by exogenous $H_2O_2$ was not observed. But various antioxidants suppressed the expression of LPS-induced IL-8 and IL-$1{\beta}$ mRNA expression of PBMC and the suppressive activity was most prominant when the pretreatment was done with TMTU. Conclusion: Oxygen free radical may have some role in the expression of IL-8 and IL-$1{\beta}$ mRNA of PBMC but that radical might not be $H_2O_2$.