Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.
Non-alcoholic fatty liver disease accompanies the rise in the prevalence of obesity, diabetes and the tendency toward high-fat dietary habits. Specifically, the higher prevalence of non-alcoholic fatty liver disease in men and postmenopausal women seems to be caused by the protective effects of estrogen against liver fibrosis, or lack thereof. There are no effective preventive therapies for liver diseases because the mechanisms underlying the progression of fatty liver diseases to chronic liver diseases and the protective effects of estrogen against fibrogenesis remain unclear. Recently, it has been reported that the hedgehog signaling pathway plays an important role in the progression of chronic liver diseases. Hedgehog, a morphogen regulating embryonic liver development, is expressed in injured livers but not in adult healthy livers. The level of hedgehog expression parallels the stages of liver diseases. Hedgehog induces myofibroblast activation and hepatic progenitor cell proliferation and leads to excessive liver fibrosis, whereas estrogen inhibits the activation of hepatic stellate cells to myofibroblasts and prevents liver fibrosis. Although the mechanism underlying the opposing actions of hedgehog and estrogen on liver fibrosis remain unclear, the suppressive effects of estrogen on the expression of osteopontin, a profibrogenic extracellular matrix protein and cytokine, and the inductive effects of hedgehog on osteopontin transcription suggest that estrogen and hedgehog are associated with liver fibrosis regulation. Therefore, further research on the estrogen-mediated regulatory mechanisms underlying the hedgehog-signaling pathway can identify the mechanism underlying liver fibrogenesis and contribute to developing therapies for preventing the progression of fibrosis to chronic liver diseases.
Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
KSBB Journal
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v.21
no.3
/
pp.204-211
/
2006
For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.
Journal of the Korea Institute of Building Construction
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v.9
no.4
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pp.63-73
/
2009
Recently, the use of lightweight panels in building structures has been increasing. Of the various lightweight panel types, styrofoam sandwich panels are inexpensive and are excellent in terms of their insulation capacity and their constructability. However, sandwich panels that include organic material are quite vulnerable to fire, and thus can numerous casualties in the event of a fire due to the lack of time to vacate and their emission of poisonous gas. On the other hand, lightweight foamed concrete is excellent, both in terms of its insulation ability and its fire resistance, due to its Inner pores. The properties of lightweight concrete is influenced by foaming agent type. Accordingly, this study investigates the insulation properties by foaming agent type, to evaluate the possibility of using light-weight foamed concrete instead of styrene foam. Our research found thatnon-heating zone temperature of lightweight foamed concrete using AP (Aluminum Powder) and FP (animal protein foaming agent) are lower than that of light-weight foamed concrete using AES (alkyl ether lactic acid ester). Lightweight foamed concrete using AES and FP satisfied fire performance requirements of two hours at a foam ratio 50, 100. Lightweight foamed concrete using AP satisfied fire performance requirements of two hours at AP ratio 0.1, 0.15. The insulation properties were better in closed pore foamed concrete by made AP, FP than with open pore foamed concrete made using AES.
This study was carried out to compare the physicochemical characteristics and preference as a sensory quality of Sogokju (Korean traditional rice wine) from waxy rice varieties. The protein and moisture contents of milled waxy rice varieties were ranged 6.9~7.2% and 12.1~ 12.6%, respectively. Nunbora had the largest grain size. In pasting properties, Hangangchalbyeo had the highest peak, trough and final viscosities, and Dongjinchalbyeo had the lowest viscosity curve. These differences suppose to be caused by the amylopectin(AP) structure: Dongjinchalbyeo has the largest short AP chains (degree of polymerization (DP) 6-12) and the smallest middle AP chains (DP 13-24) in 9 waxy rice varieties, while Hangangchalbyeo has the smallest short AP chains and the largest middle AP chains. The alcohol contents of Sogokju brewed from 9 waxy rice varieties were 17.6~19.9%. The brix degree were ranged $20.5{\sim}23.9^{\circ}Bx$. The organic acid of Sogokju consisted mainly of succinic acid, and the free sugar of it consisted mostly of glucose. The sensory evaluation showed the highest palatability at the Sogokju from Baegseolchalbyeo. The palatability was positively correlated with the brix degree, the glucose content, and the turbidity, and negatively correlated with the production yield of Sogokju.
Journal of the Korean Society of Food Science and Nutrition
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v.37
no.4
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pp.465-471
/
2008
Physicochemical characteristics of black garlic were analyzed. Colorimetry measurement showed that the black garlic, compared with fresh and steamed garlics, was the highest in a value and the lowest in L and b values. Crude lipid, crude protein, and total sugars were the highest in black garlic, which was followed by steamed and fresh garlic. On the other hand, moisture content was the lowest in the black garlic and the highest in the fresh garlic. The pH of garlics was ca. 6.8, 6.5, and 4.4 in fresh, steamed, and black garlic, respectively, which indicated that garlics tended to be acidified with the thermal processing. Total pyruvate and total thiosulfinates were the lowest in steamed garlic ($77{\mu}mol$/g and 0.07 OD/g for each) and the highest in black garlic ($278{\mu}mol$/g and 0.77 OD/g). Arabinose and galactose were detected only in black garlic and their contents were 1.6 and 13 mg/100 g, respectively. Free sugars such as glucose, sucrose and fructose were the highest in the order of fresh, steamed, and black garlic. Potassium was a predominant mineral in all garlics, constituting 76% of total minerals. Glutamic acid, arginine, and aspartic acid were the major composition amino acids in all garlics, regardless of processing conditions. 15 kinds of free amino acids were detected in fresh and steamed garlic, while five more free amino acids, O-phosphoethanolamine, and urea were additionally detected in black garlic.
Eggs are an important source of protein for the human diet. Consumers want fresher, more delicious and more sanitary eggs. In Korea, the Shell Egg Grading system (EGS) was employed in 2001. The portion of graded shell eggs has increased every year, but graded shell eggs account for only 6% of all eggs. The EGS should satisfy producers, distributors and consumers. However, the EGS does not have an official function because of many problems. Consumers cannot select various graded shell eggs in the market, and producers do not receive enough profit even though they produce top-quality graded shell eggs. There are few studies on the EGS, Therefore, this study was performed to improve the EGS. We surveyed the EGS, GP Centers and farmers. Large companies (farmers) are more satisfied than small companies with the EGS. There was a high tendency for the companies (farmers) that are not involved with the EGS to think that graded and ungraded shell eggs are similar, in contrast to the companies (farmers) connected to the EGS. We should have to change the grading system of grade shell eggs, establish of the cold chain system, change of the law for the school meals, minimize payment for the grading shell eggs for developing EGS. Based on this study, the egg industry can benefit through the improvement of the EGS.
Background: Although pulmonary resection is the standard approach for the management of pulmonary metastases from soft tissue sarcoma, most of them are unresectable and chemotherapy remains the only option. The effectiveness of the cytotoxic drugs may be limited by the toxicities that occur before the therapeutic dose is reached. The regional administration of doxorubicin using pulmonary arterial perfusion in a rodent model can produce 10 to 25 times higher concentrations in the lung than systemic administration with minimal systemic toxicities. However, it is unclear whether a high concentration of doxorubicin has beneficial effects for killing cancer cells. Material and Method: We studied this to evaluate the dose-dependent cytotoxic and apoptotic effects of doxorubicin on methylcholanthrene-induced rat fibrosarcoma(MCA) cells. This study examined the cytotoxicity and apoptosis-related gene expressions(Fas, FasL, Bax, caspase 1, caspase 2, caspase 8, Bcl-2, Bcl-xL, Bcl-xS) in MCA cells after 24 hours exposure to various concentrations of doxorubicin such as 1, 5, 10, 50, and 100 $\mu$M. Result: Dose-dependent cytotoxicity was observed after 24 hours exposure to doxorubicin. However, peak apoptosis after 24 hours exposure was observed at 5 $\mu$M of doxorubicin. Above 5 $\mu$M, apoptotic activity was decreased with dose-increment. All mRNA levels of apoptosis-related genes after 24 hours exposure were up-regulated above the control level at 1 $\mu$M of doxorubicin and then decreased by doxorubicin dose-increment except caspase 8, which showed higher levels than the control level at 5 $\mu$M. Apoptosis-related protein levels were highest at 1 $\mu$M of doxorubicin and then decreased by doxorubicin dose-increment. However, Bax and Bcl-xL proteins steadily showed higher levels than the control throughout the different concentrations of doxorubicin. Conclusion: These results suggest that apoptosis is the main cytotoxic mechanism in low concentrations of doxorubicin in MCA cells and apoptosis-related genes, such as Bax, caspase 8, and Bcl-xL, are involved. At high concentrations, doxorubicin still can kill MCA cells, even when apoptosis is inhibited, and have its propriety for achieving much cytotoxicity against MCA cells.
In order to determine the antigenic localization in the tissues of the adult Metagonimus yokegawai, immunogoldlabeling method was applied using serum immunoglobulins (IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size: 12 nm) , It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.
Park, Chan Sung;Park, Kyung Soo;Kim, Mi Lim;Kong, Hyun Joo;Yang, Kyung Mi
Journal of Life Science
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v.23
no.2
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pp.228-236
/
2013
This study was aimed at investigating whether dietary therapy using medicinal enzyme powder is effective in reducing constipation caused by loperamide in rats. Nine-week-old male Sprague Dawley were subdivided into 4 groups: normal diet group (C), loperamide treatment and normal diet (CL), medicinal enzyme powder diet (E), and loperamide treatment and medicinal enzyme powder diet (EL). Constipation was induced by subcutaneous injection of loperamide (1.5 mg/kg) 3 days prior to sacrifice. The treatment with loperamide led to an increase in weight gain, a decrease in the number and wet weight of fecal pellets, and a decrease in intestinal motility. The administration of the medicinal enzyme powder significantly reduced weight gain but increased intestinal mobility compared with the loperamide-treated group. The treatment with loperamide in the normal diet group reduced the activities of both suggesting that constipation may be involved in the low level of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT). Additionally, the loperamide treatment in the medicinal enzyme powder diet group increased the level of GOT, but reduced the level of GPT. Loperamide treatment also reduced cholesterol and increased the atherogenic index (AI) and cardiac risk factors (CRFs). Interestingly, the treatment with the medicinal enzyme powder effectively attenuated both the increase in AI and the reduction in high density lipopretein (HDL)-cholesterol, caused by the treatment with loperamide. Although there were no significant differences in the blood protein level, including hemoglobin and hematocrit, between the normal diet group and the loperamide-treated group, the administration of the medicinal enzyme powder to the loperamide-treated group effectively increased the levels of both hemoglobin and hematocrit. Collectively, the results demonstrate that the medicinal enzyme powder can help to combat the negative events caused by constipation.
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