소 모색관련 유전자 MC1R 의 RCR - RFLP Marker 를 이용한 한우육 판별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.4

The melanocortin 1 receptor(MClR) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat colour variations within several species including cattle. This study was performed to develop the identification technique of Hanwoo meat using MC1R gene associated with the coat colors of cattle. Alleles of the MC1R locus were detected by PCR-RFLP analysis and genotype frequency and DNA sequences of MC1R gene were compared among cattle breeds. Genomic DNA was extracted from meat or blood samples of five breeds including Hanwoo(n=200), Holstein(n=100), Angus(n=20), Hereford(n=20) and Charolais(n=20). The MC1R gene was used to amplify 739bp and 173bp of the bovine E-locus corresponding to positions 228-966 and 318-490, respectively, using the two specific primers. The amplified products were digested with Bse118 I or Msp I and Aci I enzymes, and DNA fragments were separated by gel electrophoresis for RFLP genotype analysis. Six genotypes, E^D/E^D E^D/E^+, E^D/e, E^+/E^+,E^+/e and e/e, controlled by three alleles E^D, E^+ and e were observed in MC1 locus. When the amplified DNA product(739bp) was digested with Bse118 I enzyme, Hanwoo meat showed a single band of 739bp, whereas two fragments of 531bp and 208bp were detected in Holstein meat and Angus breed, respectively. Also, in the RFLP patterns using Msp I enzyme, Hanwoo meat produced two fragments of 535bp and 174bp, while three fragments of 328bp, 207bp and 174bp were observed in Holstein meat and Angus breeds, respectively. Therefore, breed-specific RFLP markers showing distinct differences between these breeds were found by PCR-RFLP analysis. When the amplified DNA product(173bp) was digested with Aci I enzyme to classify subtype of E allele, the E^D allele produced three fragments of 97, 68 and 8bp, while the E^+ and d alleles produced two fragments of 173 and 8bp according to the Aci I recognition sequence. Among the six genotypes, two genotypes of E^+/e and e/e were observed in Hanwoo and their frequencies were 0.07 and 0.93, respectively. However, the E^D/ED and E^D/e genotypes were present in Holstein and E^D/E^D, E^D/E^+ and E^D/e genotypes in Angus breeds. Therefore, the E^+/e and e/e genotypes observed in Hanwoo and E^D/E^D, E^D/E^+ and E^D/e genotypes detected only in Holstein and Angus breeds may be useful as breed-specific DNA markers for distinguishing between Hanwoo meat and Holstein and Angus meats. When comparing MC1R sequences among Hanwoo, Holstein and Angus, a Gly → Val amino acid change due to a single base(G) deletion at colon 104 was found in Hanwoo. Consequently, breed specific RFLP genotypes of MC1R gene related to bovine coat colors could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

Survival of tissue-resident memory T cells requires exogenous lipid uptake and metabolism

Pan, Youdong,Tian, Tian,Park, Chang Ook,Lofftus, Serena Y.,Mei, Shenglin,Liu, Xing,Luo, Chi,O’Malley, John T.,Gehad, Ahmed,Teague, Jessica E.,Divito, Sherrie J.,Fuhlbrigge, Robert,Puigserver, Pere,Kru Macmillan Publishers Limited, part of Springer Nat 2017 Nature Vol.543 No.7644

Tissue-resident memory T (T<SUB>RM</SUB>) cells persist indefinitely in epithelial barrier tissues and protect the host against pathogens. However, the biological pathways that enable the long-term survival of T<SUB>RM</SUB> cells are obscure. Here we show that mouse CD8<SUP>+</SUP> T<SUB>RM</SUB> cells generated by viral infection of the skin differentially express high levels of several molecules that mediate lipid uptake and intracellular transport, including fatty-acid-binding proteins 4 and 5 (FABP4 and FABP5). We further show that T-cell-specific deficiency of Fabp4 and Fabp5 (Fabp4/Fabp5) impairs exogenous free fatty acid (FFA) uptake by CD8<SUP>+</SUP> T<SUB>RM</SUB> cells and greatly reduces their long-term survival in vivo, while having no effect on the survival of central memory T (T<SUB>CM</SUB>) cells in lymph nodes. In vitro, CD8<SUP>+</SUP> T<SUB>RM</SUB> cells, but not CD8<SUP>+</SUP> T<SUB>CM</SUB> cells, demonstrated increased mitochondrial oxidative metabolism in the presence of exogenous FFAs; this increase was not seen in Fabp4/Fabp5 double-knockout CD8<SUP>+</SUP> T<SUB>RM</SUB> cells. The persistence of CD8<SUP>+</SUP> T<SUB>RM</SUB> cells in the skin was strongly diminished by inhibition of mitochondrial FFA β-oxidation in vivo. Moreover, skin CD8<SUP>+</SUP> T<SUB>RM</SUB> cells that lacked Fabp4/Fabp5 were less effective at protecting mice from cutaneous viral infection, and lung Fabp4/Fabp5 double-knockout CD8<SUP>+</SUP> T<SUB>RM</SUB> cells generated by skin vaccinia virus (VACV) infection were less effective at protecting mice from a lethal pulmonary challenge with VACV. Consistent with the mouse data, increased FABP4 and FABP5 expression and enhanced extracellular FFA uptake were also demonstrated in human CD8<SUP>+</SUP> T<SUB>RM</SUB> cells in normal and psoriatic skin. These results suggest that FABP4 and FABP5 have a critical role in the maintenance, longevity and function of CD8<SUP>+</SUP> T<SUB>RM</SUB> cells, and suggest that CD8<SUP>+</SUP> T<SUB>RM</SUB> cells use exogenous FFAs and their oxidative metabolism to persist in tissue and to mediate protective immunity.

Staged development of long-lived T-cell receptor αβ T_H17 resident memory T-cell population to <i>Candida albicans</i> after skin infection

Park, Chang Ook,Fu, Xiujun,Jiang, Xiaodong,Pan, Youdong,Teague, Jessica E.,Collins, Nicholas,Tian, Tian,O'Malley, John T.,Emerson, Ryan O.,Kim, Ji Hye,Jung, Yookyung,Watanabe, Rei,Fuhlbrigge, Robert C Elsevier 2018 The Journal of allergy and clinical immunology Vol.142 No.2

<P><B>Background</B></P> <P> <I>Candida albicans</I> is a dimorphic fungus to which human subjects are exposed early in life, and by adulthood, it is part of the mycobiome of skin and other tissues. Neonatal skin lacks resident memory T (T<SUB>RM</SUB>) cells, but in adults the <I>C albicans</I> skin test is a surrogate for immunocompetence. Young adult mice raised under specific pathogen-free conditions are naive to <I>C albicans</I> and have been shown recently to have an immune system resembling that of neonatal human subjects.</P> <P><B>Objective</B></P> <P>We studied the evolution of the adaptive cutaneous immune response to <I>Candida</I> species.</P> <P><B>Methods</B></P> <P>We examined both human skin T cells and the <I>de novo</I> and memory immune responses in a mouse model of <I>C albicans</I> skin infection.</P> <P><B>Results</B></P> <P>In mice the initial IL-17–producing cells after <I>C albicans</I> infection were dermal γδ T cells, but by day 7, αβ T<SUB>H</SUB>17 effector T cells were predominant. By day 30, the majority of <I>C albicans</I>–reactive IL-17–producing T cells were CD4 T<SUB>RM</SUB> cells. Intravital microscopy showed that CD4 effector T cells were recruited to the site of primary infection and were highly motile 10 days after infection. Between 30 and 90 days after infection, these CD4 T cells became increasingly sessile, acquired expression of CD69 and CD103, and localized to the papillary dermis. These established T<SUB>RM</SUB> cells produced IL-17 on challenge, whereas motile migratory memory T cells did not. T<SUB>RM</SUB> cells rapidly clear an infectious challenge with <I>C albicans</I> more effectively than recirculating T cells, although both populations participate. We found that in normal human skin IL-17–producing CD4<SUP>+</SUP> T<SUB>RM</SUB> cells that responded to <I>C albicans</I> in an MHC class II–restricted fashion could be identified readily.</P> <P><B>Conclusions</B></P> <P>These studies demonstrate that <I>C albicans</I> infection of skin preferentially generates CD4<SUP>+</SUP> IL-17–producing T<SUB>RM</SUB> cells, which mediate durable protective immunity.</P>

Characterization of Interface States in MOS Systems by Using Photonic High-Frequency Capacitance-Voltage Responses

S.J.Song,H.T.Kim,S.S.Chi,M.S.Kim,W.S.Chang,S.D.Cho,H.T.Shin,T.E.Kim,H.J.Kang,D.J.Kim,D.M.Kim 한국물리학회 2002 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.41 No.6

Based on the photonic high-frequency capacitance-voltage response of Metal-Oxide-Semiconductor capacitors, we report an improved characterization method for the analyzing of interface states in MOS systems. An optical source with a photonic energy $E_{ph}$ = 0.943 eV ($\lambda$ = 1314.5 nm) is employed for photonic deep-depletion (fast sweep rate) high-frequency Capacitance-Voltage (photonic DD HF-CV) characterization of interface states distributed in the photo-responsive energy band. Using the photonic DD HF-CV characterization, we obtained a U-shaped distribution of $D_{it}$ over ($E_V + E_g/2 - q\phi_f) < E_t < (E_V + E_g/2 - q\phi_f + q\phi_S$) for N-type Metal-Oxide-Semiconductor capacitors and ($E_C - E_g/2 - q\phi_f + q\phi_S) < E_t < (E_C - E_g/2 - q\phi_f$) for P-type Metal-Oxide-Semiconductor capacitors.

Seroprevalence of Toxoplasma gondii and Trichinella spiralis infections in wild boars (Sus scrofa) in Korea

Kang, S.W.,Doan, H.T.T.,Noh, J.H.,Choe, S.E.,Yoo, M.S.,Kim, Y.H.,Reddy, K.E.,Nguyen, T.T.D.,Van Quyen, D.,Nguyen, L.T.K.,Kweon, C.H.,Jung, S.C. Elsevier 2013 Parasitology international Vol.62 No.6

Toxoplasma gondii and Trichinella spiralis are important zoonotic pathogens with worldwide distributions. In Korea, several outbreaks of human toxoplasmosis and trichinellosis due to the consumption of infected wild animals have been reported. The purpose of this study was to determine the seroprevalence of T. gondii and T. spiralis infections in wild boars killed in Korea from December 2009 to October 2011. A total of 521 wild boars hunted in eight provinces were examined for antibodies to T. gondii and T. spiralis by using commercial ELISA kits. Overall, 25.1% of serum samples from individual boars were seropositive for T. gondii and 1.7% were seropositive for T. spiralis. Seropositive for T. gondii was found in the boars in all the eight provinces investigated and for T. spiralis in four provinces. This is the first report on the seroprevalence of T. gondii and T. spiralis infections in wild boars in Korea. The consumption of undercooked wild boar meat may expose humans to a high risk of infection.

Impact of thermal and chemical treatment on the mechanical properties of E110 and E110G cladding tubes

M. Kir aly,Z. Hozer,M. Horvath,T. Novotny,E. Perez-Fero,N. Ver 한국원자력학회 2019 Nuclear Engineering and Technology Vol.51 No.2

The mechanical and corrosion behavior of the Russian zirconium fuel cladding alloy E110, predominantlyused in VVERs, has been investigated for many decades. The recent commercialization of a new, optimizedE110 alloy, produced on a sponge zirconium basis, gave the opportunity to compare the mechanicalproperties of the old and the new E110 fuel claddings. Axial and tangential tensile test experiments were performed with samples from both claddings in theMTA EK. Due to the anisotropy of the cladding tubes, the axial tensile strength was 10e15% higher thanthe tangential (measured by ring tensile tests). The tensile strength of the new E110G alloy was 11%higher than that of the E110 cladding at room temperature. Some samples underwent chemical treatment e slight oxidation in steam or hydrogenation e or heattreatment e in argon atmosphere at temperatures between 600 and 1000 C. The heat treatment duringthe oxidation had more significant effect on the tensile strength of the claddings than the oxidation itself,which lowered the tensile strength as the thickness of the metal decreased. The hydrogenation of thecladding samples slightly lowered the tensile strength and the samples but they remained ductile even atroom temperature.

E-commerce Adoption in Selected Agribusiness Micro, Small and Medium Retail Enterprises (MSMEs) in Laguna, Philippines

Loida E. Mojicaa,Jannie Gracelyn P. Avilesb,Jeanette Angeline B. Madambac,Agnes T. Banzond 아시아무역학회 2018 Journal of Asia Trade and Business Vol.4 No.2

Agribusiness has to be at par with other businesses in adapting to the globalized market environment to take advantage of the new opportunities brought about by the new market reach. With the increased internet and social media penetration in the Philippines as shown in the digital statistical indicators in 2016 and 2017, and the support of telecommunication service providers, opportunities for adoption of e-commerce are available for agribusiness companies.E-commerce becomes increasingly important to business organizations as businesses connect with the other players in the supply chain. Although e-commerce has been found to benefit businesses, and there is awareness among agribusiness MSMEs of these benefits, adoption of e-commerce agribusiness retail micro, small and medium Enterprises (MSMEs) in Laguna, Philippines is relatively slow compared to big businesses. The traditional way of doing business, specifically, the personalized interaction with the customers that the MSMEs would like to maintain, the generally small business size, the cost of investing, implementing and maintaining e-commerce infrastructure and people and the lack of knowledge of e-commerce among agribusiness retail MSMEs were the barriers to e-commerce adoption among retail MSMEs.

AFLP Marker를 이용한 한국 재래돼지의 유전적 다양성 및 품종식별

정의룡,김우태,김연수,이정구,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.6

본 연구는 한국재래돼지의 순수 혈통정립 및 유전자원 보존을 목적으로 AFLP 다형성을 분석하고 이를 유전적 표지인자로 이용하여 재래돼지의 유전적 특성을 분자 수준에서 규명하고 품종 집단내 유전적 변이성과 품종 특이적인 DNA marker를 탐색하며 동시에 타품종들과의 유전적 근연관계를 추정하기 위해 수행하였다. 13종류의 selective primer 조합형을 이용하여 분석한 결과 총 band의 수는 611개로 각 조합형 당 평균 47개의 band가 확인되었으며 이 가운데 다형적 band가 152개로 다형성 수준은 약24.5%였다. 한국 재래돼지의 다형율과 유전적 다양성 값은 각각 29.8%와 2.9로 개량종에 비해서 다소 높은 경향이었다. 한편, 13종류의 primer 조합형 가운데 E35/T38 및 E38/H13 primer 조합형에서 품종간의 차이를 나타내는 DNA marker가 확인되었다. 특히, 재래돼지에서 검출된 E35/T38 primer 조합형의 0.50kb, 0.25kb 및 0.38kb의 DNA band는 개량종과 명확히 구별되어 이들 품종 특이적 DNA band는 한국 재래돼지의 품종식별에 유용한 표지인자로 이용 가능할 것으로 기대된다. 품종간 유전적 근연관계에서 한국 재래돼지는 Hampshire종과 유전적 유사성이 가장 높은 것으로 추정되었다. 본 연구에서 검출한 한국 재래돼지의 AFLP 유전자 지문은 재래돼지 집단의 유전적 변이성 및 타 품종과의 근연관계 분석뿐만 아니라 경제형질과 연관된 marker 개발에 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다. For the purpose of genetic conversation and utilization of Korean native pig(KNP) as a valuable animal genetic resource, DNA polymorphisms of AFLP (amplified fragment length polymorphisms) as genetic markers were analyzed in KNP and foreign pig breeds (Landrace, Duroc, Hamphsire and Yorkshire). Using these AFLP markers, the genetic structure and characteristics of KNP population were analyzed at the DNA level and the genetic variability and diversity within and between breed population were evaluated. Breed-specific DNA marker for KNP was screened, and phylogenetic relationship within and among breeds were estimated. A total of 611 AFLP markers were amplified by 13 selective primer combinations, and the average number of bands per primer combination was 47.0. Among them 152 bands were polymorphic (24.5%). The rate of polymorphisms and genetic diversity values of KNP (29.8% and 2.90) was higher than these of other foregin breeds. E35/T38 and E38/H13 primer combinations produced AFLP banding patterns which clearly discriminated between KNP and other foreign pig breeds. Three bands(0.50kb, 0.25kb and 0.20kb) identified in E35/T38 and two bands(0.45kb and 0.38kb) identified in E38/H13 primer combinations were present in all of the KNP examined, but not present in the foreign pig breeds. Therefore, these two primer combinations could be used as breed-specific DNA markers for breed identification of KNP. In comparison of genetic distances among pig breeds, KNP was the most closely related to the Hampshire breed. AFLP fingerprints may be useful for genetic variability and relationships and development of breed-specific DNA markers in pig breeds.

PCR - SSCP 기법을 이용한 소 MC1R 유전자의 다형성 분석 및 한우육 감별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.1

The single-strand conformation polymorphism(SSCP) is a point mutation screening method based on changes in the secondary structure in a defined single-stranded DNA fragment caused by a change in sequence. This study was performed to develop the identification technique of Hanwoo meat using PCR-SSCP marker of MC1R gene associated with the coat colors of cattle. Hanwoo, Holstein and imported cattle breeds(Angus, Hereford and Charolais) were used for the detection of PCR-SSCP genotypes of MC1R gene. The specific primers were used to amplify a 138bp and a 390bp of the bovine E-locus(including point mutation) corresponding to positions 318-455 and 318-707, respectively, in the MC1R gene. For SSCP genotype analysis, the PCR products were denatured at 95℃ for 5min, loaded on a 12-15% nondenaturing polyacrylamide gel and detected by silver staining. When the PCR product of 138bp was separated by gel electrophoresis, two genotypes of E^+/e and e/e were detected only in Hanwoo meat, but E^D/E^D and E^D/e and E^D/E^D genotypes were observed in Holstein meat and Angus breed, respectively. Also, in the SSCP analysis using PCR product of 390bp, Hanwoo meat produced banding pattern of two fragments, while single fragment was observed in Holstein meat and Angus breed. Therefore, breed-specific SSCP markers showing distinct differences between Hanwoo and Holstein and Angus were found by PCR-SSCP analysis. This study shows that the PCR-SSCP technique is relatively simple, fast and a practical tool for determination of MC1R genotypes of cattle breeds when compared with PCR-RFLP method. Therefore, the PCR-SSCP markers of MC1R gene could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

Effects of Resveratrol and Essential Oils on Growth Performance, Immunity, Digestibility and Fecal Microbial Shedding in Challenged Piglets

Ahmed, S.T.,Hossain, M.E.,Kim, G.M.,Hwang, J.A.,Ji, H.,Yang, C.J. Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.5

A study was conducted to evaluate the effects of resveratrol and essential oils from medicinal plants on the growth performance, immunity, digestibility, and fecal microbial shedding of weaned piglets. A total of 48 weaned piglets (8 kg initial weight, 28-d-old) were randomly allotted to four dietary treatments with 3 replications of 4 piglets each. The dietary treatments were NC (negative control; basal diet), PC (positive control; basal diet+0.002% apramycin), T1 (basal diet+0.2% resveratrol), and T2 (basal diet+0.0125% essential oil blend). All piglets were orally challenged with 5 ml culture fluid containing $2.3{\times}10^8$ cfu/ml of Escherichia coli KCTC 2571 and $5.9{\times}10^8$ cfu/ml Salmonella enterica serover Typhimurium. The PC group (p<0.05) showed the highest average daily gain (ADG) and average daily feed intake (ADFI) throughout the experimental period, although feed conversion ratio (FCR) was improved in the T1 group (p>0.05). Serum IgG level was increased in the T1 group, whereas TNF-${\alpha}$ levels was reduced in the supplemented groups compared to control (p<0.05). The PC diet improved the dry matter (DM) digestibility, whereas PC and T2 diets improved nitrogen (N) digestibility compared to NC and T1 diets (p<0.05). Fecal Salmonella and E. coli counts were reduced in all treatment groups compared to control (p<0.05). Fecal Lactobacillus spp. count was increased in the T2 group compared to others (p<0.05). Dietary treatments had no significant effect on fecal Bacillus spp. count throughout the entire experimental period. Based on these results, resveratrol showed strong potential as antibiotic alternatives for reversing the adverse effects of weaning stress on growth performance, immunity and microbial environment in E. coli and Salmonella-challenged piglets.