Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.
The present study was performed to study the genetic relationship of productive life with production and type traits of Korean Holsteins at first three lactations. The data for the analysis from 56,054, 28,997, and 11,816 animals of first, second and third parity cows which were born from 2006 to 2011 were collected by Dairy Cattle Improvement Center, National Agricultural Co-operative Federation. Milk, protein and fat yields adjusted for 305 days and average somatic cell score considered as production traits and analyzed type traits were stature, strength, body depth, dairy form, rump angle, rump width, rear leg side view, foot angle, front attachment placement, rear attachment height, rear attachment width, udder cleft, udder depth, front teat placement and front teat length. A multi trait genetic analysis was performed using Wombat program with restricted maximum likelihood animal model composed of fixed effect of birth year, farm and the random effect of animal and random residual effect according to the traits. Heritability estimates of productive life were between 0.06 and 0.13. Genetic and phenotypic correlations between production and productive life traits ranged from 0.35 to 0.04 for milk, 0.16 to 0.05 for protein and 0.18 to 0.02 f 15-0034 (2nd) 150520 or fat. Somatic cells score showed a negative genetic and phenotypic correlation with productive life and also udder type traits, indicating that the selection for higher udder traits will likely to improve resistance to mastitis and persistence in the herd. Among all dairy form type traits, udder characters such as udder cleft showed a significant relationship with productive life. However, a specific change of heritabilities or correlations were not observed with the change of parity. Moreover, further studies are needed to further confirm the significance of the above traits and the effect of parity on above relationships in order to minimize both voluntary and involuntary culling rates while improving herd health and maintaining high yielding dairy cows.
Journal of the Korea Academia-Industrial cooperation Society
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v.15
no.11
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pp.6518-6526
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2014
The aim of this study was to obtain a complete picture of green catalytic technology developments. The data set for analysis was collected from the data from 4,172 patents, 28,726 technical papers and 548 Government R&D projects. The covering periods were 2001 to 2012 for patents and technical papers, whereas while that for the Government R&D projects were 2001 to 2011. The analysis methodologies consisted of qualitative and quantitative approaches. The patent for catalytic technology has been increasing, even though it was developed some time ago. The increase in patents for catalytic technology has been outstanding since 2008. As a result of the analysis of patents, fuel cell comprised 41.9%, followed by coal liquefaction 23.6%. The analysis of technical papers ranked fuel cell ranked 1st, followed by hydrogen. Fuel cells, hydrogen and coal liquefaction outperformed in green catalytic technological developments. As a result of this study, it was apparently concluded that the speed for the green economy from the fossil fuel-based economy has accelerated. Therefore it will be necessary to prepare for the early realization of a green energy-based economy on a variety of aspects as soon as possible.
Ulmus davidiana var. japonica Rehder (Urticales: Ulmaceae) (UD) is a tree widespread in northeast Asia. It is traditionally used for anticancer and anti-inflammatory therapy. The present study investigated the effect of an ethanol extract of UD on vascular tension and its underlying mechanism in rats. The dried root bark of UD was ground and extracted with 80% ethanol. The prepared UD extract was used in further analysis. The effect of UD on the cell viability, vasoreactivity and hemodynamics were investigated using propidium iodide staining in cultured cells, isometric tension recording and blood pressure analysis, respectively. Low dose of UD ($10{\sim}100{\mu}g/ml)$ did not affect endothelial cell viability, but high dose of UD reduced cell viability. UD induced vasorelaxation in the range of $0.1{\sim}10{\mu}g/ml$ with an $ED_{50}$ value of $2{\mu}g/ml$. UD-induced vasorelaxation was completely abolished by removal of the endothelium or by pre-treatment with L-NAME, an inhibitor of nitric oxide synthase. UD inhibited calcium influx induced by phenylephrine and high $K^+$ and also completely abolished the effect of L-NAME. Intravenous injection of UD extracts (10~100 mg/kg) decreased arterial and ventricular pressure in a dose-dependent manner. Moreover, UD extracts reduced the ventricular contractility (+dP/dt) in anesthetized rats. However, UD-induced hypotensive actions were minimized in L-NAME-treated rats. Taken together, out results showed that UD induced vasorelaxation and has antihypertensive properties, which may be due the activation of nitric oxide synthase in endothelium.
To obtain basic information on the potential use of Camellia japonica leaf as a raw material in functional food, leaf antioxidant and anticancer activities were investigated. The radical-scavenging activity of various solvent fractions from the leaf, as shown by the DPPH radical test, increased in a dose-dependent manner, with the water fraction showing the highest activity. The reducing power of various solvent fractions from the leaf was also dose-dependent, and, again, the water fraction showed the highest reducing power. The water fraction showed strong antioxidant activity in the linoleic acid test and was also capable of scavenging nitrite in a dose-dependent manner. Proportions of 92.15% and 95.61% of available nitrite were scavenged by the water and butanol fractions, respectively, at levels of $1,000{\mu}g/mL$. Both butanol and water fractions exhibited strong inhibitory effects on the growth of human lung and colon cancer cells. The total phenolic contents of the butanol and water fractions were 216.26 mg/g and 220.68 mg/g, respectively. High-performance liquid chromatography (HPLC) showed that quercetin and epicatechin were the predominant phenolic compounds in the water fraction. The activities of this fraction are attributable to the presence of these phenolic compounds, particularly quercetin and epicatechin.
Kim, Kyoung-Woon;Cho, Mi-La;Lee, Sang-Heon;Min, So-Youn;Park, Mi Kyung;Park, Sung-Hwan;Jue, Dae-Myung;Kim, Ho-Youn
IMMUNE NETWORK
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v.3
no.4
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pp.310-319
/
2003
Inflammatory mediators has been recognized as an important role in the pathogenesis of rheumatoid arthritis (RA). IL-17 is increasingly recognized as an important regulator of immune and inflammatory responses, including induction of proinflammatory cytokines and osteoclastic bone resorption. Evidence of the expression and proinflammatory activity of IL-17 has been demonstrated in RA synovium and in animal models of RA. However, the signaling pathways that regulate IL-17 production remain unknown. In the present study, we investigated the role of the phosphatidylinositol 3 kinase (PI3K)-Akt pathway in the regulation of IL-17 production in RA. PBMC were separated from RA (n=24) patients, and stimulated with various agents (anti CD3, anti CD28, PHA, ConA, IL-15). IL-17 levels were determined by sandwich ELISA and RT-PCR. The production of IL-17 was significantly increased in cells treated with anti-CD3 antibody, PHA, IL-15 or MCP-1 (P<0.05). ConA also strongly induced IL-17 production (P<0.001), whereas TNF-alpha, IL-1beta, IL-18 or TGF-beta did not. IL-17 was detected in the PBMC of patients with osteoarthritis (OA) but their expression levels were much lower than those of RA PBMC. Anti-CD3 antibody activated the PI3K-Akt pathway and activation of the PI3K-Akt pathway resulted in a pronounced augmentation of nuclear factor kappaB ($NF-{\kappa}B$). IL-17 production by activated PBMC in RA is completely or partially blocked in the presence of $NF-{\kappa}B$ inhibitor PDTC and PI3K-Akt inhibitor, wortmannin and LY294002, respectively. Whereas the inhibition of AP-1 and extracellular signal-regulated kinase (ERK)1/2 did not affect IL-17 production. These results provide new insight into that PI3K/Akt and $NF-{\kappa}B$ dependent signal transduction pathway could be involved in the overproduction of key inflammatory cytokine, IL-17 in rheumatoid arthritis.
This study was conducted to improve quality of takju where natural honey is used to control the ripening fermentation and the amount of sediment derived from takju mash. A koji was prepared using rice starch and Aspergillus awamori var. kawachii. Takju mash was prepared by alcohol fermentation with Saccharomyces cerevisiae and a 3-step addition of steamed rice. The clean part of the mash (CPM) was separated from the sediment at $5^{\circ}C$ and 5% (w/v) of natural honey was added and then ripened for 23 days at 5, 10, or $15^{\circ}C$. Temperature, pH, acidity, and total sugar content showed no significant differences, but a 0.2 percent reduction in alcohol content occurred during storage. However, CPM ripened with honey had a comparatively higher score on sensory evaluation than did immature CPM with added honey added. Takju with 8% alcohol content was prepared by mixing the water from the mixed CPM ripened with honey together with 100, 50, and 25% of the frozen sediment. Several quality characteristics of the takju were checked over 37 days of fermentation at $10^{\circ}C$. The pH was sustained between 4.1 and 4.3, and changes in the number of viable yeast cells, acidity, total sugar amounts, and alcohol content showed similar patterns but differences in scale. Smaller amounts of sediment affected the stability of the takju. Mixing the CPM ripened with natural honey at low temperature moderately reduced the amount of sediment in the mash and resulted in a highly flavorful takju with an extended shelf life.
Background: An easy, reproducible and simple marker is needed to estimate phase of endometrial pathologic lesions such as hyperplasia and endometrial cancer and distinguish from pathologically normal results. We here aimed to clarify associations among neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), endometrial hyperplasia and cancer in patients with abnormal uterine bleeding. Materials and Methods: Patients (n=161) who were admitted with abnormal uterine bleeding and the presence of endometrial cells on cervical cytology or thick endometrium were investigated. The study constituted of three groups according to pathologic diagnosis. Group 1 included endometrial precancerous lesions like hyperplasia (n=63), group 2 included endometrial cancerous lesions (n=38) and group 3 was a pathologically normal group (n=60). Blood samples were obtained just before the curettage procedure and the NLR was defined as the absolute neutrophil count divided by the absolute lymphocyte count; similarly, PLR was defined as the absolute platelet count divided by the absolute lymphocyte count. Results: The white blood cell count was significantly higher in patients with cancer than in those with hyperplasia (p=0.005). The platelet count and neutrophil to lymphocyte ratio were significantly higher in patients with cancer than in control patients, but there was significantly no difference between patients with hyperplasia and other groups (p=0.001 and p=0.025 respectively). PLR was significantly lower in control subjects than in other groups (p<0.001), but there was no significant difference between patients with hyperplasia and those with cancer. Conclusions: PLR was significantly lower in control subjects than in other groups. Thus both hyperplasia and cancer may be differentiated from pathologically normal patients by using PLR. White blood cell count was significantly higher in patients with cancer than in those with hyperplasia and pathologically normal patients. Therefore white blood cell count may be used for discriminate hyperplasia to cancer. By using multiple inflammation parameters, discrimination may be possible among endometrial cancer, endometrial precancerous lesions and pathologically normal patients.
Kim, Dong Hyun;Oh, Sung Yong;Kim, So Yeon;Lee, Seul;Koh, Myeong Seok;Lee, Ji Hyun;Lee, Suee;Kim, Sung-Hyun;Park, Heon Soo;Hur, Won Joo;Jeong, Jin Sook;Ju, Mi Ha;Seol, Young Mi;Choi, Young-Jin;Chung, Joo Seop;Kim, Hyo-Jin
Asian Pacific Journal of Cancer Prevention
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v.15
no.24
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pp.10985-10989
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2015
Background: The capability for DNA double-strand breaks (DSBs) repair is crucial for inherent radiosensitivity of tumor and normal cells. We have investigated the clinicopathologic significance of DNA repair gene expression in nasopharyngeal (NP) carcinoma. Materials and Methods: A total of 65 NP cancer patients who received radiotherapy were included. The immunopositivity to Ku 70, DNA-PKcs, MRN, RAD50, XRCC4, and LIG4 were examined in all tumor tissues. Results: The patients comprised 42 males and 23 females, with a median age of 56 years (range, 18-84). The expression levels of RAD50 (0,+1,+2,+3) were 27.7%, 32.3%, 21.5%, and 18.5%. LIG4 (${\pm}$) were 43.1% and 56.9% respectively. The 5-year OS rate of patients with LIG4 (${\pm}$) were 90% and 67.9%, respectively (p=0.035). The 5-year TTP rate of patients with LIG4 (${\pm}$) were 75.9%, 55.5%, respectively (P=0.039). Conclusions: Our results suggest the possibility of predicting the radiosensitivity of NP cancer by performing immunohistochemical analysis of LIG4.
Fusarium graminearum (teleomorph Gibberella zeae) is an important plant pathogen that causes head blight of major cereal crops such as wheat, barley, and rice, as well as causing ear and stalk rot on maize worldwide. Plant diseases caused by this fungus lead to severe yield losses and accumulation of harmful mycotoxins in infected cereals [1]. Fungi utilize spore production as a mean to rapidly avoid unfavorable environmental conditions and to amplify their population. Spores are produced sexually and asexually and their production is precisely controlled. Upstream developmental activators consist of fluffy genes have been known to orchestrate early induction of condiogenesis in a model filamentous fungus Aspergillus nidulans. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we characterized functions of the F. graminearum fluffy gene homologs [2]. We found that FlbD is conserved regulatory function for conidiogenesis in both A. nidulans and F. graminearum among five fluffy gene homologs. flbD deletion abolished conidia and perithecia production, suggesting that FlbD have global roles in hyphal differentiation processes in F. graminearum. We further identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum [3]. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. F. graminearum ortholog of Aspergillus nidulans wetA has been shown to be involved in conidiogenesis and conidium maturation [4]. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidia dormancy by suppressing microcycle conidiation in F. graminearum. In A. nidulans, FlbB physically interacts with FlbD and FlbE, and the resulting FlbB/FlbE and FlbB/FlbD complexes induce the expression of flbD and brlA, respectively. BrlA is an activator of the AbaA-WetA pathway. AbaA and WetA are required for phialide formation and conidia maturation, respectively [5]. In F. graminearum, the AbaA-WetA pathway is similar to that of A. nidulans, except a brlA ortholog does not exist. Amongst the fluffy genes, only fgflbD has a conserved role for regulation of the AbaA-WetA pathway.
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