In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.
In this study, the radioprotective effects of Grifola umbellata hot water extracts (Gu-extract) on mice were investigated. Single pre-administration of Gu-extract increased the 40-day survival ratio of irradiated mice from 65.5% to 78.6%. The growth of 3 week old male mice in the irradiated group was slightly retarded as compared to those of the control and Gu-extract treated mice. The average spleen and thymus weights of the irradiated mice were lower than those of the control and Gu-extract treated mice. The weight reduction of testis in the irradiated mice was significant. While it was relatively slight in the Gu-extract treated mice as compared to that of control mice. No significant difference in the weight was observed in heart, kidney or liver among three groups. The leukocytes of the Gu-extract treated mice did not decrease dramatically as in the irradiated group, but recovery patterns were similar in both groups. Reduction of erythrocytes were similar in both groups but its recovery occurred more rapidly in the extract treated group. The glucose level of the Gu-extract treated group did not change during the period examined, while it was still higher in the irradiated group than the level in the control group in two weeks. The cholesterol levels in the irradiated and the Gu-extract treated groups were higher than that of control group on day 7, but decreased to the level of the control group on day 14. No difference was observed in total protein amount of the serum among the three groups. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the livers of the irradiated and extract treated groups, in which some proteins were missing or less present.
The objective of this study was to model the kinetics of S. aureus survival on high risk foods in school foodservice operations. After inoculating S. aureus ATCC25923 onto the various high risk foods, the effects of competitive microorganism, storage temperatures($25^{\circ}C$, $35^{\circ}C$), and initial contamination levels ($1.0{\times}10^2\;CFU/g$, $1.0{\times}10^5\;CFU/g$) on the growth of S. aureus were investigated. Lag time decreased and specific growth rate increased with a storage temperature ($25^{\circ}C$<$35^{\circ}C$) and with a higher initial inoculation level ($1.0{\times}10^2\;CFU/g$<$1.0{\times}10^5\;CFU/g$). Previously it was shown that S. aureus is a weaker competitor than other organisms, but it proliferates aggressively in a noncompetitive environment. However, in our study, when S. aureus was used to inoculate japchae (glass noodles with sauteed vegetables) and meat ball, the growth of S. aureus was similar and more active with competitive organisms than that without competitive organisms. Regardless of other factors, the initial level of S. aureus was a more significant factor of the growth. High inoculation levels of S. aureus were reached at 6 log CFU/g within 3 hours. An incubation temperature of $35^{\circ}C$ and the animal protein component of menu items also were identified as significant factors influencing the growth of S. aureus. Therefore, the duration of time meals are stored before serving should be considered a critical control point. Food service providers must control time and temperature to insure the safety of cooked foods.
Ixeris dentata var. albiflora Nakai, a herbal plant, is often used to make a strong stomach as an antiphlogistic used when dyspepsia and to improve appetite in Korea and China. And also it is used for adult diseases such as diabetes and liver diseases as Korean traditional medicine. In this study, the composition and DPPH radical scavenging activities of the root of Ixeris dentata var. albiflora Nakai and its effects on cell viability on vero and chang cells were investigated. Moisture, crude ash, crude protein and crude lipid were 79.14, 2.49, 8.28 and 2.56 g/100 g respectively. The highest mineral content was K. The major free sugars were glucose, fructose and sucrose. Major fatty acid are linoleic acid, palmic acid and linolenic acid. Major amino acids were glutamic acid, arginine and aspartic acid and the total contents of amino acids were 28.12 mg/g. The methanol extracts were further fractionated with n-hexane, chloroform, ethylacetate, butanol and water to get an active fraction. In addition, cell viabilities in each fraction were determined. Methanol extract, butanol, and aqueous fraction showed strong survival rates in vero cell and chang cell viability test, and hexane, chloroform, and ethylacetate fraction were examined for toxin in a cell. The root of Ixeris dentata var. albiflora Nakai had scavenging activities against DPPH radicals in a dose-dependent assay. Ethylacetate fraction's SC50 was $6.8\{\mu}g/mL$, very strong DPPH radical scavenging activities, but water fraction did not show any activity.
Kim Kyoung-Duck;Kang Yong-Jin;Lee Hae-Young;Kim Kang-Woong;Kim Kyong-Min;Lee Sang-Min
Journal of Aquaculture
/
v.19
no.3
/
pp.173-177
/
2006
This study was conducted to evaluate extruded pellets (EP) for growth of adult flounder by comparing with raw fish-based moist pellet (MP). Two replicate groups of 150 fish per each tank (initial mean weight 329 g) were fed one of seven EP (EP1, EP2, EP3, EP4, EP5, EP6 and EP7) and a MP for 8 months. Survival of fish fed the MP was not significantly different from that of fish fed the EP1, EP5 and EP7, but significantly higher than that of fish fed the EP2, EP3, EP4 and EP6 (P<0.05). Weight gain of fish fed the MP was significantly lower than that of fish fed the EPI (P<0.05), but not significantly different from that of fish fed EP2, EP3, EP4, EP5, EP6 and EP7. Feed efficiency of fish fed the MP was significantly lower than EP1, EP3, EP4, EP5 and EP6 (P<0.05), but not significantly different from that of fish fed EP2 and EP7 Feed supply (kg/tank) of fish fed the MP was significantly higher than that of fish fed all EP (P<0.05). Condition factor of fish fed the MP was not significantly different from that of fish fed all EP. The contents of moisture, crude protein and lipid in dorsal muscle and whole body was not significantly different among the groups. It is concluded that the dietary formulation used in the EP1, EP3, TP4, EP5 and EP6 can be applied in the practical extruded pellet feeds for adult flounder (329-680g).
Song Si Yeol;Lee Sang-wook;Yu Eunsil;Cho Kyung-Ja;Park Jin-hong;Kim Sang Yoon;Nam Soon Yuhl;Lee Bong-Jae;Kim Sung Bae;Choi Seung-Ho;Ahn Seung Do;Shin Seong Soo;Choi Eun Kyung
Radiation Oncology Journal
/
v.23
no.1
/
pp.1-8
/
2005
Purpose : The objective of this study was to determine whether the expressions of the two components of DNA-dependent protein kinase, Ku70 and DNA-PKcs, influence the response to radiotherapy (RT) and outcome of treatment of non-disseminated nasopharyngeal carcinoma (NPC) in patients who received definitive RT. Materials and Methods : Sixty-six patients with NPC who were treated with radiotherapy alone or with concurrent chemotherapy between June 1995 and December 2001 were divided into groups based on the levels of immunoreactivity for Ku70 and DNA-PKcs in pretreatment biopsy specimens. The over-expression of Ku70 or DNA-PKcs groups Included patients whose biopsy specimens showed at least 50% immunopositive tumor cells; patients in which less than 50% of the tumor cells in the biopsy tissues were immunopositive were placed in the low Ku70 and DNA-PKcs groups. The immunoreactivities for Ku70 and DNA-PKcs were retrospectively compared with the sensitivity of the tumor to radiation and the patterns of therapy failure. Univariate analyses were peformed to determine the prognostic factors that influenced locoregional control of NPC. Results : The five-year locoregional control rate was significantly higher in the low Ku70 group (Ku(-)) (85%) than in the high Ku70 group (Ku(+)) (42%) (p=0.0042). However, there were no differences in the metastases-free survival rates between the two groups (Ku70 (+), 82%; Ku70 (-), 78%; p=0.8672). Univariate analysis indicated that the over-expression of Ku70 surpassed other well-known predictive clinocopathologic parameters as an Independent prognostic factor for locoregionai control. Eighteen of 22 patients who had locoregional recurrences of the tumor displayed an over-expression of Ku70. No significant association was found between the level of DNA-PKcs expression and the clinical outcome. Conclusion : Our data suggest that the level of Ku70 expression can be used as a molecular marker to predict the response to RT and the locoregional control after RT and concurrent chemotherapy in patients with non-disseminated NPC.
In this study, we carried out an experiment for estimation the larval digestibility in aspects which digestive enzymatic activities and nutrition of the rotifers, Brachionus rotundiformis. Thus we enhanced the digestive enzymatic activity through the addition of starch for the increase of digestibility of rotifer (starch-rotifer), and compared with the feed efficiency through rearing of the olive flounder, Paralichthys olivaceus used rotifer lipid-enriched with Algamac $2000^{(R)}$ (CE-rotifer). The digestive enzyme activities (except for TG-lipase), total protein contents, total essential amino acid, essential amino acids (methionin and phenylalanine) of starch-rotifer (the rotifer used a starch as additive, and enriched not) was assayed significantly higher than CE-rotifer (P<0.05). And total lipid, lipid classes (except for sterol) and fatty acids as DHA and EPA showed higher in CE-rotifer than starch-rotifer (P<0.05). But, sterol contents and ST/TG ratio were shown significantly higher in starch-rotifer (P<0.05). The flounder larvae supplied the two rotifers showed standard length and body weight that not significantly differed with ranges $3.72{\sim}3.79\;mm$ and $32.9{\sim}37.8\;mg$/larva on 6 days after hatching (DAH), respectively (P>0.05). However, these of 12 DAH showed the values of significantly higher to $5.94{\pm}0.249\;mm$, $144.0{\pm}23.86\;mg$/larva and $26.2{\pm}12.13%$ in standard length, body weight and survival in CE-flounder than that of starch-flounder (P<0.05). The hydrolytic enzymatic activities of flounder larvae severally supplied the two rotifers showed the significantly higher activities in acidic -amylase, neutral -amylase, TG-lipase, lysozyme and acidic phosphatase in starch-flounder on 5 DAH (P<0.05). But neutral $\alpha$-amylase, three proteases and two phosphatases of CE-flounder on 11 DAH showed the significantly higher activities than that of starch-flounder (P<0.05). Therefore, for the flounder, Paralichthys olivaceus larvae just depleted yolk was more beneficial to supply the feed, rotifer, enhanced the digestibility than to supply the feed lipid-enriched for aspect of larval digestibility up to 6 DAH, thereafter nutrition of absorption due to the development of digestive organs suggested that enrichment effect appeared with larval somatic growth. Consequently, investigation more detailed about the larval digestive physiological and nutritional requirement variations after 6 DAH will be necessary, thereafter.
Lim, Eun Gyeong;Kim, Guen Tae;Kim, Bo Min;Kim, Eun Ji;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
Journal of Life Science
/
v.26
no.6
/
pp.663-672
/
2016
The Cnidium monnieri (L.) Cusson is an annual plant distributed in China and Korea. The fruit of C. monnieri is used as a medicinal herb that is effective for the treatment of carbuncle and pain in female genitalia. However, the anti-cancer effects of CME have not yet been reported. In this study, we assessed the apoptotic effects and cell cycle arrest effects of ethanol extracts from C. monnieri on HCT116 colon cancer cells. The results of an MTT assay and LDH assay demonstrated a decrease in cell viability and the cytotoxic effects of CME. In addition, the number of apoptotic body and the apoptotic rate were increased in a dose-dependent manner through Hoechst 33342 staining and Annexin V-PI double staining. In addition, cell cycle arrest occurred at the G1 phase by CME. Protein kinase B (Akt) plays an important role in cancer cell survival, growth, and division. Akt down-regulates apoptosis-mediated proteins, such as mammalian target of rapamycin (mTOR), p53, and Glycogen Synthase kinase-3β (GSK-3β). CME could regulate the expression levels of p-Akt, p-mTOR, p-GSK-3β, Bcl-2 family members, caspase-3, and PARP. Furthermore, treatment with CME, LY294002 (PI3K/Akt inhibitor), BIO (GSK-3β inhibitor), and Rapamycin (mTOR inhibitor) showed that apoptotic effects occurred through the regulation of the AKT/mTOR/GSK-3β signaling pathway. Our results demonstrated CME could induce apoptosis and cell cycle arrest in HCT116 colon cancer cells.
Osteosarcoma (OS) is the most common and malignant bone tumors. Although many types of resection surgery and experimental agents were developed, median survival and clinical prognosis are poorly investigated. Recently, several researches have reported that Eucheuma cottonii has potent as protective effects of coal dust-induced lung damage via inhibition of malondialdehyde (MDA) and oxidative stress in bronchoalveolar lavage fluids (BALF). However, anti-cancer effects and specific molecular mechanism of extract from Eucheuma cottonii (EE) has not been clearly studied yet. This study evaluated that anti-cancer potential of EE in human osteosarcoma Saos-2 cells. EE indicated cytotoxicity on Saos-2 cells in a dose-dependent manner. Morphological degradation and nucleic condensation were also observed under the EE treatment. However, it did not significantly affect on non-cancerous kidney HEK-293 cells under the same concentration which is shown cytotoxicity on Saos-2 cells. The phosphorylation of Fas-Associated Death Domain (FADD) and expression of cleaved caspase-8, -7 and -3 were upregulated in a dose-dependent manner. In immunofluorescence staining, expression level of Fas and cleaved PARP were upregulated by EE treatment. Furthermore, treatment of EE induces upregulation of sub G1 phase by flow cytometry analysis. The results demonstrated that EE has a therapeutic potential against osteosarcoma via FADD mediated caspase cascade apoptosis signal pathway.
Kim, Jong-Hyeon;Kim, Jong-Suk;Yoo, Wan-Hee;Hur, Hyeon
Journal of Food Hygiene and Safety
/
v.21
no.4
/
pp.213-217
/
2006
The halophilic bacterium Vibrio vulnificus is known to be a foodborne pathogen that causes septicemia in human. V. vulnificus infection is characterized by the high fatality rates and the primary attack against a person who have underlying diseases such as liver cirrhosis. However, there is no effective treatment for V. vulnificus septicemia except for classical treatments such as antibiotics. Recently, it has been known that lipoprotein (LDL) plays a major role in the protection against infection and inflammation. Consequently in this paper we analyzed the effects of LDL on V. vulnificus septicemia. We purified V. vulnificus cytolysin, a major virulent factor of V. vulnificus infection and measured inhibitory effects of mouse serum, cholesterol, and LDL on its hemolytic activity. Next experiments were performed to investigate whether LDL has a protective role against septicemia induced by V. vulnificus in mice. Intraperitoneal injection of LDL (1mg as protein) into mice 3hr before V. vulnificus $(1\times10^6\;CFU)$ injection, and V. vulnificus -induced lethality was determined. For the determination the relationship between LDL or cholesterol and prognosis, we determined serum levels of cholesterol and lipoprotein from V. vulnificus septicemia patients (n=15) who had visited the Chonbuk National University Hospital in Chonju. V. vulnificus cytolysin -induced hemolysis of mice erythrocytes was completely inhibited by serum, cholesterol, and low-density lipoprotein. V. vulnificus- induced lethality of mice injected with LDL showed only 40% compared to 100% of control. In survival groups (n=4) of V. vulnificus septicemia patients (n=15), their serum LDL and cholesterol revealed normal levels ($153.3{\pm}40.7,\;LDL;\;190.8{\pm}16.3$, Total cholesterol). However, in death groups (n=11) showed very low levels ($35.6{\pm}13.9,\;LDL;\;59.2{\pm}15.1$, Total cholesterol). Our study indicates that cholesterol and LDL are a prognosis indicator of V. vulnificus septicemia as well as an inhibitor of virulent action of V. vulnificus cytolysin. We suggested that the serum levels of cholesterol or LDL would be major index in the treatment and prevention of V. vulnificus septicemia.
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