• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.179-188
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• 2017

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.187-190
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• 2004

Protein kinase C exists as a family of serine/threonine kinases which are broadly classified into three groups as cPKC nPKC and aPKC depending on their cofactor requirements. Previous studies have shown that the role of PKC in the process of mouse oocyte maturation. For example, phorbol 12-myristate 13-acetate which is known as an activator of cPKC and nPKC inhibits germinal vesicle break down and 1st polar body extrusion in maturing oocytes. In this study, the effect of thymeleatoxin, a specific activator of cPKC not nPKC, was tested comparing with PMA to address the roles of cPKC and nPKC during mouse oocyte maturation. Cumulus-oocyte complex were cultured in M16 medium for 6 or 12 hr with each of these PKC activators to investigate the effect of germinal vesicle breakdown (GVBD) or the extrusion of 1st polar body. IC/sup 50/ of GVBD were at concentrations of 50nM in PMA and 400nM in thymeleatoxin and of 1st polar body extrusion were 20nM in PMA and 200nM in thy- meleatoxin. The results suggest that activation of nPKC is more closely related to the inhibition of GVBD and 1st polar body extrusion than activation of cPKC. Additionally, we found that the oocytes inhibited 1st polar body extrusion with PMA or thymeleatoxin were arrested in metaphase I of first meiosis.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.307-311
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• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG ($33.1{\pm}9.6$ vs $21.7{\pm}8.3%$). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP-treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The mRNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.287-294
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• 2011

Autophagy is a process of intracellular bulk protein degradation, in which the accumulated proteins and cytoplasmic organelles are degraded. It plays important roles in cellular homeostasis, apoptosis, and development, but its role during early embryo development remains contentious. Therefore, in the present study, we investigated the effects of 3-methyladenine (3-MA) on early embryonic development in pigs, we also investigated several indicators of developmental potential, including mitochondrial distribution, genes expressions (autophagy-, apoptosis- related genes), apoptosis and ER-stress, which are affected by 3-MA. After in vitro maturation and fertilization, presumptive pig embryos were cultured in PZM-3 medium supplemented with 3-MA for 2 days at $39^{\circ}C$ 5% $CO_2$ in air. Developmental competence to the blastocyst stage in the presence of 3-MA was gradually decreased according to increasing concentration. Thus, all further experiments were performed using 2 mM 3-MA. Blastocysts that developed in the 3-MA treated group decreased LC3-II intensity and expressions of autophagy related genes than those of the untreated control, resulting in down-regulates the autophagy. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 3-MA treated group compared with control ($6.0{\pm}1.0$ vs $3.3{\pm}0.6$, p<0.05). Also, the expression of the pro-apoptotic gene Bax increased in 3-MA treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. Mito Tracker Green FM staining showed that blastocysts derived from the 3-MA treated group had lower mitochondrial integrity than that of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. Then, the expression of the spliced form of pXBP-1 product (pXBP-1s) increased in 3-MA treated group, resulting increase of ER-stress. Taken together, these results indicate that inhibition of autophagy by 3-MA is closely associated with apoptosis and ER-stress during preimplantation periods of porcine embryos.

• Journal of Animal Science and Technology
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• v.60 no.2
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• pp.1.1-1.6
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• 2018

Background: Maize fodder is being used as staple feed for livestock but it lacks protein and essential amino acids; lysine and tryptophan. Intercropping maize with leguminous soybean crop is promising technique under limited land resources of South Korea but it can only give considerable advantages when adequate distance is provided between corn and soybean rows. Main aim of present study was to find-out adequate distance between corn and soybean seeding rows for optimum growth, yield and nutritive value of intercropped forage. Methods: Different interrow distances between corn and soybean were evaluated under four treatments, viz. 1) Corn sole as positive control treatment 2) Zero cm between corn and soybean (control); 2) Five cm between corn and soybean; 3) 10 cm between corn and soybean, with three replicates under randomized block design. Results: Findings depicted that height and number of corn stalks and ears were similar (P > 0.05) among different treatments. Numerically average corn ear height was decreased at zero cm distance. Dry matter percentage in all components; corn stalk, corn ear and soybean was also found not different (P > 0.05) but dry matter yield in component of corn ear was lower (P < 0.05) at zero cm distance as compared to that of 5 and 10 cm interrow distances. In case of nutritive value, total digestible nutrient yield in intercropped corn was also found lower (P < 0.05) at zero cm distance than that of 5 and 10 cm interrow distances between corn and soybean seeding rows. Substantial decrease in dry matter yield of maize ear at zero cm distance might be attributed to factor of closed interrow spacing which made interplant competition more intensified for light interception, necessary for photosynthetic activity. Lower dry matter yield in ear also reduced total digestible nutrients in intercropped maize because it was determining factor in calculation of digestible nutrients. The optimum yield and nutritive value of forage at wider interrow distance i.e. 5 cm between corn and soybean might be due to adequate interseed distance. Conclusion: Conclusively, pattern of corn and soybean seeding in rows at 5 cm distance was found suitable which provided adequate interrow distance to maintain enough mutual cooperation and decreased competition between both species for optimum production performance and nutritive value of intercropped forage.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.152-157
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• 2004

Gluten was extracted from domestic wheat flour using UTH buffer (4 M urea in 0.1 M Tris-HCl, pH 8.6) and validated by SDS-PAGE analysis for production of wheat flour products with reduced gluten content.. Anti-gluten polyclonal antibody was made by administering extracted gluten fraction on animal model. Anti-gluten serum titer of extracted gluten fraction was evaluated by ELISA, and that of antibody titer according to administration period. Anti-gluten sera were used for ELISA and immunoblot analysis before and after hydrolysis of gluten fraction at optimal pH and temperature condition for each protease. Gluten fraction separated by SDS-PAGE showed several bands covering 75 to 10 kDa, in which anti-gluten sera were 25, 34, and 45 kDa. Enzyme hydrolysis of gluten fraction revealed protein band sizes to be lower than 15 kDa. Content of pretense from bovine pancreas (b.p. protease) for gluten hydrolysis was estimated as 1 mg in 10 mL gluten fraction extracted for 4 hr.

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.2
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• pp.121-126
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• 1999

Manure management is a top priority on both dairy and swine farrns at present. This experiment was carried out to determine the effect of different types and N rates of liquid manure on forage yield and quality of oats(Avena sativa L.) in Suweon, 1997. Seven treatments consisting of chemical fertilizer $120kg\;N\;ha^{-1}$, liquid cattle manure 120, 240 and $360kg\;N\;ha^{-1}$, and liquid swine manure 120, 240 and $360kg\;N\;ha^{-1}$ were arranged in a randomized complete block design with three replications. As increasing N rate of the liquid manure, the plant height was increased, but dry matter content of oats was decreased as increasing liquid manure N rate. There were significant differences in dry matter yield of oats among the liquid manure N rates(P<0.05). Crude protein(CP) content of oats ranged from 14.7 to 24.1% (P<0.05), and CP yield was orderly ranked as liquid swine manure $360kg\;N\;ha^{-1}$>liquid cattle manure $360kg\;N\;ha^{-1}$>liquid swine manure $240kg\;N\;ha^{-1}$(P<0.05). Acid detergent fiber(ADF) content of the liquid cattle manure treatment showed a slightly higher than that of the liquid swine manure treatment(P<0.05). Neutral detergent fiber(NDF) content of the liquid cattle manure treatment was lower than that of the liquid swine manure treatment(P<0.05). Relative feed value(RFV) of the liquid cattle manure treatment decreased with increasing liquid manure N rates compared with that of the liquid swine manure treatment. Based on the results of this experiment, it is indicates that to increase the N rates from liquid manure application could be obtained from the increase of forage yield.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.6
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• pp.837-843
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• 2001

Two experiments were conducted to compare the feed intake patterns and growth performance of Meishan and Yorkshire growing pigs. Experiment 1 was carried out over a 6-wk period and used 48 barrows with equal numbers of purebred Meishan (M) and Yorkshire (Y). Pigs were allocated to four groups of 12 pigs consisting of equal numbers of M and Y. Initial BW were $36.4{\pm}0.32kg$ and $42.1{\pm}1.41kg$ for M and Y, respectively. Experiment 2 was carried out over a 5-week period and used 48 pigs consisting of equal numbers of both barrows and gilts and of crossbred Meishan$\times$Yorkshire (MY) and purebred Yorkshire (Y) animals. Pigs were allotted to 6 pens of 8 pigs, with 4 single- and 2 mixed-genotype groups (initial $BW=28.5{\pm}0.99kg$). In both experiments, pigs were given ad libitum access to a grower diet (17% crude protein, 0.9% lysine, 3365 kcal/kg ME) via feed intake recording equipment (F.I.R.E.). Pigs carried an ear-tag transponder with an unique identification which allowed the time, duration, and size of individual meals to be recorded. In Exp. 1, Y had higher ADG (721 vs 353 g, p<0.01), daily feed intake (DFI; 2.338 vs 1.363 kg, p<0.01), made more frequent visits to the feeder per day (NFV; 18.5 vs 7.7, p<0.01), had a shorter feeder occupation time per visit (FOV; 7.4 vs 12.9 min, p<0.01), and ate less feed per visit (FIV; 130 vs 177 g, p<0.01) than M pigs. Feed consumption rates (CR) were greater for Y compared to M (19.3 vs 14.8 g/min, p<0.01). Feeder occupation time per day (FOD) was longer for Y than M (114.3 vs 82.8 min/pig, p<0.01). Yorkshire pigs visited the feeder more frequently between 0800 and 1100 h. Meishan pigs showed more frequent feeder visits between 0600 and 0800 h, and between 1600 and 2100 h when feeding competition with Y was reduced. In Exp. 2, there was no effect of genotype or group composition on DFI, ADG or gain:feed ratio. Crossbred pigs (MY) made fewer feeder visits (12.6 vs 17.7, p<0.01), and had greater FIV (124 vs 98 g/visit, p<0.01), and longer FOV (8.11 vs 7.24 min/visit, p<0.01) and FOD (112 vs 100 min, p<0.05) than Y pigs. Results of this study suggest substantial genetic variation in feeding patterns as well as in growth performance.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.2
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• pp.288-298
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• 2016

Resveratrol acts as a free radical scavenger and a potent antioxidant in the inhibition of numerous reactive oxygen species (ROS). The function of resveratrol and resveratrol-loaded nanoparticles in protecting human lung cancer cells (A549) against hydrogen peroxide was investigated in this study. The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay was performed to evaluate the antioxidant properties. Resveratrol had substantially high antioxidant capacity (trolox equivalent antioxidant capacity value) compared to trolox and vitamin E since the concentration of resveratrol was more than $50{\mu}M$. Nanoparticles prepared from ${\beta}$-lactoglobulin (${\beta}$-lg) were successfully developed. The ${\beta}$-lg nanoparticle showed 60 to 146 nm diameter in size with negatively charged surface. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. Fluorescein isothiocynate-conjugated ${\beta}$-lg nanoparticles were identified into the cell membrane of Caco-2 cells, indicating that nanoparticles can be used as a delivery system. Hydrogen peroxide caused accumulation of ROS in a dose- and time-dependent manner. Resveratrol-loaded nanoparticles restored $H_2O_2$-induced ROS levels by induction of cellular uptake of resveratrol in A549 cells. Furthermore, resveratrol activated nuclear factor erythroid 2-related factor 2-Kelch ECH associating protein 1 (Nrf2-Keap1) signaling in A549 cells, thereby accumulation of Nrf2 abundance, as demonstrated by western blotting approach. Overall, these results may have implications for improvement of oxidative stress in treatment with nanoparticles as a biodegradable and non-toxic delivery carrier of bioactive compounds.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.1
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• pp.64-70
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• 2017

Objective: In the present study, role of increasing levels of Ecklonia cava (seaweed) supplementation in diets was investigated on growth performance, coefficient of total tract apparent digestibility (CTTAD) of nutrients, serum immunoglobulins, cecal microflora and intestinal morphology of weanling pigs. Methods: A total of 200 weaned pigs (Landrace${\times}$Yorkshire${\times}$Duroc; initial body weight $7.08{\pm}0.15kg$) were randomly allotted to 4 treatments on the basis of body weight. There were 5 replicate pens in each treatment including 10 pigs of each. Treatments were divided by dietary Ecklonia cava supplementation levels (0%, 0.05%, 0.1%, or 0.15%) in growing-finishing diets. There were 2 diet formulation phases throughout the experiment. The pigs were offered the diets ad libitum for the entire period of experiment in meal form. Results: The pigs fed with increasing dietary concentrations of Ecklonia cava had linear increase (p<0.05) in the overall average daily gain, however, there were no significant differences in gain to feed ratio, CTTAD of dry matter and crude protein at both phase I and phase II. Digestibility of gross energy was linearly improved (p<0.05) in phase II. At day 28, pigs fed Ecklonia cava had greater (linear, p<0.05) Lactobacillus spp., fewer Escherichia coli (E. coli) spp. (linear, p<0.05) and a tendency to have fewer cecal Clostridium spp. (p = 0.077). The total anaerobic bacteria were not affected with supplementation of Ecklonia cava in diets. Polynomial contrasts analysis revealed that villus height of the ileum exhibited a linear increase (p<0.05) in response with the increase in the level of dietary Ecklonia cava. However, villus height of duodenum and jejunum, crypt depth, villus height to crypt depth ratio of different segments of the intestine were not affected. Conclusion: The results suggest that Ecklonia cava had beneficial effects on the growth performance, cecal microflora, and intestinal morphology of weanling pigs.