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Effects of Supplementation of Ruminally Protected Amino Acids on In vitro Ruminal Parameters and Milk Yield and Milk Composition of Dairy Cows in Mid-lactation (보호아미노산의 추가 공급이 반추위 발효성상 및 비유중기 착유우의 유량 및 유성분에 미치는 영향)

  • Lee, Jong-Min;Nam, In-Sik;Ahn, Jong-Ho
    • Journal of Animal Science and Technology
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    • v.50 no.2
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    • pp.199-208
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    • 2008
  • This study was undertaken to investigate the effects of ruminally protected amino acids (Methionine and Lysine) on in vitro ruminal parameters, and in vivo milk yield and milk composition in mid-lactating cows. In the first in vitro experiment, there were no statistical significances between treatments in ruminal pH and dry matter digestibility during various incubation times. In the second in vivo experiment, milk yield decreased by 11.92% in control and 5.68% in the treatment respectively, but decrease rate of milk yield in the treatment was lower than control. Milk yields naturally decreased as time goes by since the DIMs(Days in milk) of the cows in experiment were in mid-lactation period. 4% FCM(Fat corrected milk) and milk protein yields also, respectively, decreased by 11.25% and 11.09% in control and 6.16% and 5.47% in the treatment as compared with the intial. Milk protein and milk fat production were higher in the treatment(0.90kg, 1.10kg) than those of control(0.66kg, 0.79kg). Milk fat content significantly increased with supplementing protected amino acids as compared to control(P<0.05). From the above results, protected amino acids were positively utilized in the performances of mid-lactating cows without inhibiting rumen fermentation. Further investigation is suggested for essential amino acid composition and intestinal digestion rate out of rumen bypass protein in dietary protein to be estimated.

A Reliable Protocol for transfection of mature primary hippocampal neurons using a neuron-glia co-culture system (신경세포-신경교세포 공동배양을 이용한 성숙한 해마신경세포의 효율적인 형질전환 방법)

  • Lee, Hyun-Sook;Cho, Sun-Jung;Jung, Yong-Wook;Jin, Ing-Nyol;Moon, Il-Soo
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.198-203
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    • 2007
  • DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

Statistical Optimization of Culture Conditions of Probiotic Lactobacillus brevis SBB07 for Enhanced Cell Growth (프로바이오틱 Lactobacillus brevis SBB07의 균체량 증가를 위한 배양 조건 최적화)

  • Jeong, Su-Ji;Yang, Hee-Jong;Ryu, Myeong Seon;Seo, Ji Won;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Journal of Life Science
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    • v.28 no.5
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    • pp.577-586
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    • 2018
  • We recently reported the potential probiotic properties of Lactobacillus brevis SBB07 isolated from blueberries. The present study investigates the effect of culture conditions such as temperature, initial pH, culture time, and medium constituent for industrial application. The ingredients of the medium to improve cell growth were selected by Plackett-Burman design (PBD) and central composite design (CCD) within a desirable range. The PBD was applied with 19 factors: seven carbon sources, six nitrogen sources, and six microelements. Protease peptone, corn steep powder (CSP), and yeast extract were found to be significant factors for the growth of SBB07. The CCD was then applied with three variables found from the PBD at five levels, and the optimum values were decided for the three variables: protease peptone, CSP, and yeast extract. In the case of the growth of SBB07, the proposed optimal media contained 2.0% protease peptone, 2.5% CSP, and 2.0% yeast extract, and the maximum dried-cell weight was predicted to be 2.93963 g/l. By the model verification, it was confirmed that the predicted and actual results are similar. Finally, the study investigated the effects of incubation temperature and initial pH at the optimized medium. It was confirmed that the dried-cell weight increased from $2.2933{\pm}0.0601g/l$ to $3.85{\pm}0.0265g/l$ when compared to the basal medium at $37^{\circ}C$ and initial pH 8.0. Establishing the optimal culture condition for SBB07 provides good potential for applications in probiotics and can serve as the foundation for the industrialization of materials.

Evaluation of Fertilization Effect of Slow-Release Complex Fertilizer on Pepper Cultivation (원예용 완효성 복합비료의 고추에 대한 시비효과 평가)

  • Lee, Chang-Hoon;Lee, Hyub;Ha, Byung-Hyun;Kang, Chang-Sun;Lee, Yong-Bok;Kim, Pil-Joo
    • Korean Journal of Environmental Agriculture
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    • v.26 no.3
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    • pp.228-232
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    • 2007
  • Slow-release fertilizers (SRF) have been used to reduce nutrient loss through increasing fertilizer efficiency and to save labor. Several SRFs were developed for rice plant in Korea, but there is few for horticultural crop plants. Two slow-release complex fertilizers, 100T and 150T, which made for controlling nitrogen release time up to 100 and 150 days, respectively, were selected for the incubation test cto evaluate nitrogen (N) release rate in soil. The N of urea selected as the control was completely released within a week after application. Sixty three and 53% of total N were released from 110T and 150T of slow release fertilizers within 8th weeks after application, respectively. For pepper cultivation CF110 and CF150, new slow-release complex fertilizer, were made of mixing 40% of conventional fertilizer and 60% of 110T and 150T, respectively, based on the amount of recommended fertilizer for pepper cultivation $(N-P_2O_5-K_2O=190-112-149\;kg\;ha^{-1})$, and were totally applied before pepper transplanting in the field as the basal fertilizer. Inorganic N $(NH_4^+-N+NO_3^--N)$ concentration in soil was higher in the CF110 treatment than in the control (NPK) at all period of pepper cultivation. In the CF150 treatment concentration of inorganic N in soil was low compared to control up to 8th weeks after transplanting. However, there was no difference in plant height and nutrient content of pepper leave between CF110 treatment and the control. In comparison, plant height was significantly lower in CF150 than the control and CF110 treatments. Around 4% of fresh pepper yield was increased in CF110 compared to the control, but it was decreased to about 2% by CF150 treatment. Conclusively, CF110 form could be recommended as a slow release fertilizer for pepper cultivation.

Characterization of Chitin Deacetylase Produced from Mucor rouxii (Mucor rouxii가 생산한 Chitin deacetylase의 특성)

  • SOHN Heung-Sik;PARK Seong-Min;SON Byung-Yil;CHOI Hyeon-Mee;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.121-126
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    • 1999
  • In order to degrade chitin by enzymatic hydrolysis, it is required from screening highly active deacetylase. To this end, we examined three fungal strains and it turned out that Mucor rouxii produced highly active deacetylase, this enzyme exhibited the highest enzymatic activity against colloidal chitin. The conditions for growing Mucor rouxii are as follows; the effective carbon source, nitrogen source, adequate initial pH, temperature and incubation time were $2\%$ glucose, $1.33\%$ yeast extract, $0.66\%$ pepton, 4.5, $25{\pm}2^{\circ}C$ and 48hr, respectively. The optimum pH and temperature for purified enzyme activity were 5.5 and $40^{\circ}C$, respectively. The purified enzyme was stable at pH ranging from 4.5 to 5.5. However, the enzyme activity was decreased to less than $50\%$ at pH blow 45 and above 7.5. At temperatures above $50^{\circ}C$, the enzyme activity was decreased remarkably. The enzyme was inhibited by LiC1, $HgCl_2$, and $BaCl_2$, but stimulated by $CaCl_2$ and $ZnC1_2$, The activity of purified enzyme was increased by L-cysteine and 2-mercaptoethanol, while decreased by O-phenanthroline, p-CMB, EDTA, and iodoacetate. The $K_m$ and the $V_{max}$ value of purified enzyme were $1.2\%$ and 59.5 U/mg, respectively. The deacetylation activity of purified enzyme was not detected at optimal reaction condition when chitin particle suspension was used.

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Characterization of a new commercial strain 'Goni' by intra-specific hyphal anastomosis in Pleurotus ostreatus (계통간 교잡에 의한 백색느타리 품종 '고니'의 육성 및 그 특성)

  • Yoo, Young Bok;Lee, Sang Cheol;Kim, Eun Jung;Kong, Won Sik;Jang, Kab Yeul;Shin, Pyung Gyun
    • Journal of Mushroom
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    • v.7 no.3
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    • pp.130-134
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    • 2009
  • To develop new white variety of Oyster mushroom, all white varieties which have been collected and kept in the lab were revived and screened their cultural characteristics. 84 intra-specific Oyster mushroom hybrids between the white-colored mutants Suhan and Wonhyeong were developed using hyphal anastomosis technique in 2007. The Po2007-63 ($2842-7{\times}0205-7$) was shown the best cultural characteristics, selected to be a new variety and named as 'Goni'. The new commercial strain, 'Goni' has white pilei and grows well under spring and autumn conditions in Korea. The fruiting bodies of 'Goni' are of an excellent quality in that not only the stipe is thick and long but also the pileus is small and hard. The optimum temperatures for mycelial growth and fruiting body development were $25-30^{\circ}C$ and $10-16^{\circ}C$, respectively. Time period required for the initiation of the first fruiting body is about 3 to 5 days depending on the temperatures. The shape of fruiting body is thin funnel shape. Fruiting body production per bottle was about $91{\pm}13$ g which is almost 97% quantity compared to that of other variety 'Miso'. Relatively low temperature incubation ($11^{\circ}C$) resulted in the development of better quality of 'Goni' mushrooms. When two different media including potato dextrose medium and mushroom complete medium were compared, the growth of mushroom were much faster in mushroom complete medium at $20-25^{\circ}C$, but not at $25^{\circ}C$. Similar results were observed with other variety 'Miso'. Analysis of the genetic characteristics of the new commercial strain 'Goni' showed a major DNA profile as that of the parental Suhan when primer URP 1 was used, but different to 'Miso' that was used as a control. When screens were performed with primer URP 2, DNA patterns were similar both to that of the parents and 'Miso'. This new variety of the white Oyster mushroom has a clean and fresh image that corresponds well to "health food". We therefore expect that this new strain will satisfy the consumers demand for variety and excellent mushrooms.

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Persistence of Fungicide Pencycuron in Soils (토양 중 살균제 Pencycuron의 잔류 특성)

  • An, Xue-Hua;An, Wen-Hao;Im, Il-Bin;Lee, Sang-Bok;Kang, Jong-Gook
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.296-305
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    • 2006
  • The adsorption and persistence of pencycuron {1-(4-chlorobenzyl) cyclopentyl-3-phenylurea} in soils were investigated under laboratory and field conditions to in order to assess the safety use and environmental impact. In the adsorption rate experiments, a significant power function of relation was found between the adsorbed amount of pencycuron and the shaking time. Within one hour following the shaking, the adsorption amounts in the SCL and the SiCL were 60 and 65% of the maximum adsorption amounts, respectively. The adsorption reached a quasi-equilibrium 12 hours after shaking. The adsorption isotherms followed the Freundlich equation. The coefficient (1/n) indicating adsorption strength and degree of nonlinearity was 1.45 for SCL and 1.68 to SiCL. The adsorption coefficients ($K_d$) were 2.31 for SCL and 2.92 to SiCL, and the organic carbon partition coefficient, $K_{oc}$, was 292.9 in SCL and 200.5 inSiCL. In the laboratory study, the degradation rate of pencycuron in soils followed a first-order kinetic model. The degradation rate was greatly affected by soil temperature. As soil incubation temperature was increased from 12 to $28^{\circ}C$, the residual half life was decreased from 95 to 20 days. Arrhenius activation energy was 57.8 kJ $mol^{-1}$. Furthermore, the soil moisture content affected the degradation rate. The half life in soil with 30 to 70% of field moisture capacity was ranged from 21 to 38 days. The moisture dependence coefficient, B value in the empirical equation was 0.65. In field experiments, the half-life were 26 and 23 days, respectively. The duration for period of 90% degradation was 57 days. The difference between SCL and SiCL soils varied to pencycuron degradation rates were very limited, particularly under the field conditions, even though the characteristics of both soils are varied.

Sphingosine-1-Phosphate-Induced Migration and Differentiation of Human Mesenchymal Stem Cells to Smooth Muscle Cells (Sphingosine-1-phosphate에 의한 중간엽 줄기세포의 이동과 평활근세포로의 분화)

  • Song, Hae-Young;Shin, Sang-Hun;Kim, Min-Young;Kim, Jae-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.183-193
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    • 2011
  • Migration and differentiation of mesenchymal stem cells are crucial for tissue regeneration in response to injury. Sphingosine-1-phosphate (S1P) is a bioactive lipid that regulates a variety of biological processes, including proliferation, survival, differentiation and motility. In the present study, we determined the role of S1P in migration and differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs). S1P stimulated migration of BMSCs in a dose- and time-dependent manner, and pre-incubation of the cells with pertussis toxin completely abrogated S1P-induced migration, suggesting involvement of Gi-coupled receptors in S1P-induced cell migration. S1P elicited elevation of intracellular concentration of $Ca^{2+}$ ($[Ca^{2+}]_i$) and pretreatment with VPC23019, an antagonist of $S1P_1/S1P_3$, blocked S1P-induced migration and increase of $[Ca^{2+}]_i$. Small interfering RNA-mediated knockdown of endogenous $S1P_1$ attenuated S1P-induced migration of BMSCs. Furthermore, S1P treatment induced expression of $\alpha$-smooth muscle actin ($\alpha$-SMA), a smooth muscle marker, and pretreatment with VPC23019 abrogated S1P-induced $\alpha$-SMA expression. S1P induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and pretreatment of cells with SB202190, an inhibitor of p38 MAPK, or adenoviral overexpression of a dominant-negative mutant of the p38 MAPK blocked S1P-induced cell migration and $\alpha$-SMA expression. Taken together, these results suggest that S1P stimulates migration and smooth muscle differentiation of BMSCs through an $S1P_1$-p38 MAPK-dependent mechanism.

A Study on Users' Resistance toward ERP in the Pre-adoption Context (ERP 도입 전 구성원의 저항)

  • Park, Jae-Sung;Cho, Yong-Soo;Koh, Joon
    • Asia pacific journal of information systems
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    • v.19 no.4
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    • pp.77-100
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    • 2009
  • Information Systems (IS) is an essential tool for any organizations. The last decade has seen an increasing body of knowledge on IS usage. Yet, IS often fails because of its misuse or non-use. In general, decisions regarding the selection of a system, which involve the evaluation of many IS vendors and an enormous initial investment, are made not through the consensus of employees but through the top-down decision making by top managers. In situations where the selected system does not satisfy the needs of the employees, the forced use of the selected IS will only result in their resistance to it. Many organizations have been either integrating dispersed legacy systems such as archipelago or adopting a new ERP (Enterprise Resource Planning) system to enhance employee efficiency. This study examines user resistance prior to the adoption of the selected IS or ERP system. As such, this study identifies the importance of managing organizational resistance that may appear in the pre-adoption context of an integrated IS or ERP system, explores key factors influencing user resistance, and investigates how prior experience with other integrated IS or ERP systems may change the relationship between the affecting factors and user resistance. This study focuses on organizational members' resistance and the affecting factors in the pre-adoption context of an integrated IS or ERP system rather than in the context of an ERP adoption itself or ERP post-adoption. Based on prior literature, this study proposes a research model that considers six key variables, including perceived benefit, system complexity, fitness with existing tasks, attitude toward change, the psychological reactance trait, and perceived IT competence. They are considered as independent variables affecting user resistance toward an integrated IS or ERP system. This study also introduces the concept of prior experience (i.e., whether a user has prior experience with an integrated IS or ERP system) as a moderating variable to examine the impact of perceived benefit and attitude toward change in user resistance. As such, we propose eight hypotheses with respect to the model. For the empirical validation of the hypotheses, we developed relevant instruments for each research variable based on prior literature and surveyed 95 professional researchers and the administrative staff of the Korea Photonics Technology Institute (KOPTI). We examined the organizational characteristics of KOPTI, the reasons behind their adoption of an ERP system, process changes caused by the introduction of the system, and employees' resistance/attitude toward the system at the time of the introduction. The results of the multiple regression analysis suggest that, among the six variables, perceived benefit, complexity, attitude toward change, and the psychological reactance trait significantly influence user resistance. These results further suggest that top management should manage the psychological states of their employees in order to minimize their resistance to the forced IS, even in the new system pre-adoption context. In addition, the moderating variable-prior experience was found to change the strength of the relationship between attitude toward change and system resistance. That is, the effect of attitude toward change in user resistance was significantly stronger in those with prior experience than those with no prior experience. This result implies that those with prior experience should be identified and provided with some type of attitude training or change management programs to minimize their resistance to the adoption of a system. This study contributes to the IS field by providing practical implications for IS practitioners. This study identifies system resistance stimuli of users, focusing on the pre-adoption context in a forced ERP system environment. We have empirically validated the proposed research model by examining several significant factors affecting user resistance against the adoption of an ERP system. In particular, we find a clear and significant role of the moderating variable, prior ERP usage experience, in the relationship between the affecting factors and user resistance. The results of the study suggest the importance of appropriately managing the factors that affect user resistance in organizations that plan to introduce a new ERP system or integrate legacy systems. Moreover, this study offers to practitioners several specific strategies (in particular, the categorization of users by their prior usage experience) for alleviating the resistant behaviors of users in the process of the ERP adoption before a system becomes available to them. Despite the valuable contributions of this study, there are also some limitations which will be discussed in this paper to make the study more complete and consistent.

Combined Effects of Filter-feeding Bivalve and Zooplankton on the Growth Inhibition of Cyanobacterium Microcystis aeruginosa (남세균 제어를 위한 동물플랑크톤(Daphnia magna)과 패류(Unio douglasiae)의 단독 및 혼합적용)

  • Kim, Nan-Young;Park, Myung-Hwan;Hwang, Su-Ok;Kim, Baik-Ho;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.48 no.2
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    • pp.108-114
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    • 2015
  • Single - and combined effects of a domestic freshwater bivalve Unio douglasiae (7.6~8.6 cm in shell length) and zooplankton Daphnia magna (1~2 mm in body size) were examined to understand whether they inhibit the growth of harmful cyanobacterial bloom (i.e. Microcystis aeruginosa) in a eutrophic lake. The experiments were triplicated with twelve glass aquaria (40 L in volume); three aquaria without mussel and zooplankton, served as a control, three zooplankton aquaria (Z, density=40 indiv. $L^{-1}$), three mussel aquaria (M, density=0.5 indiv. $L^{-1}$), and three mussel plus zooplankton aquarium (ZM, density=40 indiv.Z $L^{-1}$ plus 0.5 indiv.M/L), respectively. Algal growth inhibition (%) calculated as a difference in the concentration of chlorophyll-a (Chl-a) before and after treatment. Chl-a in all aquaria decreased with the time, while a greatest algal inhibition was seen in the ZM aquaria. After 24 hrs of incubation, Chl-a concentration at the mid-depth (ca. 15 cm) in ZM aquaria reduced by 90.8% of the control, while 63.2% and 79.8% in Z and M aquaria, respectively. Interestingly, during the same period, the surface Chl-a was diminished by 51.9% and 65.4% relative to the control in Z and ZM aquaria, while 27.4% of initial concentration decreased in M aquarium, respectively. These results suggest that 1) this domestic freshwater filter-feeding bivalve plays a significant role in the control of cyanobacterial bloom (M. aeruginosa), and 2) the combination with zooplankton and mussel has a synergistic effect to diminish them, compared to the single treatment of zooplankton and mussel.