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Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

  • Yuan, Yu-Guo;Song, Shao-Zheng;Zhu, Meng-Ming;He, Zheng-Yi;Lu, Rui;Zhang, Ting;Mi, Fei;Wang, Jin-Yu;Cheng, Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1175-1182
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    • 2017
  • Objective: To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases. Methods: TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT). Results: The targeted efficiency for BLG gene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate biallelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.

EA-D p45-IgG as a Potential Biomarker for Nasopharyngeal Carcinoma Diagnosis

  • Chen, Hao;Luo, Yao-Ling;Zhang, Lin;Tian, Li-Zhen;Feng, Zhi-Ting;Liu, Wan-Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7433-7438
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    • 2013
  • Aim: To identify new biomarkers for NPC diagnosis with an anti-EBV Western blot test kit. Methods: Serum samples from 64 NPC patients and healthy subjects with four specific VCA-IgA/EA-IgA profiles were tested with an anti-EBV Western blot test kit from EUROIMMUN AG. Proteins were quantified with scores of intensity visually assigned to the protein bands. The markers which showed statistical differences between the NPC and non-NPC subjects were further evaluated in another 32 NPC patients and 32 controls in comparison with established biomarkers including VCA-IgA, EA-IgA, EBV-related protein IgG, and EBV DNA. Results: Among the markers screened, EA-D p45-IgG showed a statistically significant difference (p < 0.05) between NPC and non-NPC subjects with VCA-IgA positivy. In 32 VCA-IgA positive NPC patients and 32 control subjects, the diagnostic accuracy of EA-D p45-IgG was 78.1% with a positive predictive value of 77.8% and a negative predictive value of 78.6%. In the verification experiment, the specificity and sensitivity of EA-D p45-IgG were 75.0% and 90.6 %, respectively. Conclusions: EA-D p45-IgG might be a potential biomarker for NPC diagnosis, especially among VCA-IgA positive subjects.

Improving Participation in Colorectal Cancer Screening: a Randomised Controlled Trial of Sequential Offers of Faecal then Blood Based Non-Invasive Tests

  • Symonds, Erin L;Pedersen, Susanne;Cole, Stephen R;Massolino, Joseph;Byrne, Daniel;Guy, John;Backhouse, Patricia;Fraser, Robert J;LaPointe, Lawrence;Young, Graeme P
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8455-8460
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    • 2016
  • Background: Poor participation rates are often observed in colorectal cancer (CRC) screening programs utilising faecal occult blood tests. This may be from dislike of faecal sampling, or having benign bleeding conditions that can interfere with test results. These barriers may be circumvented by offering a blood-based DNA test for screening. The aim was to determine if program participation could be increased by offering a blood test following faecal immunochemical test (FIT) non-participation. Materials and Methods: People were invited into a CRC screening study through their General Practice and randomised into control or intervention (n=600/group). Both groups were mailed a FIT (matching conventional screening programs). Participation was defined as FIT completion within 12wk. Intervention group non-participants were offered a screening blood test (methylated BCAT1/IKZF1). Overall participation was compared between the groups. Results: After 12wk, FIT participation was 82% and 81% in the control and intervention groups. In the intervention 96 FIT nonparticipants were offered the blood test - 22 completed this test and 19 completed the FIT instead. Total screening in the intervention group was greater than the control (88% vs 82%, p<0.01). Of 12 invitees who indicated that FIT was inappropriate for them (mainly due to bleeding conditions), 10 completed the blood test (83%). Conclusions: Offering a blood test to FIT non-participants increased overall screening participation compared to a conventional FIT program. Blood test participation was particularly high in invitees who considered FIT to be inappropriate for them. A blood test may be a useful adjunct test within a FIT program.

Assessment of lactic acid bacteria isolated from the chicken digestive tract for potential use as poultry probiotics

  • Merisa Sirisopapong;Takeshi Shimosato;Supattra Okrathok;Sutisa Khempaka
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1209-1220
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    • 2023
  • Objective: The use of probiotics as an alternative to antibiotics in animal feed has received considerable attention in recent decades. Lactic acid bacteria (LAB) have remarkable functional properties promoting host health and are major microorganisms for probiotic purposes. The aim of this study was to characterize LAB strains of the chicken digestive tract and to determine their functional properties for further use as potential probiotics in poultry. Methods: A total of 2,000 colonies were isolated from the ileum and cecal contents of the chickens based on their phenotypic profiles and followed by a preliminary detection for acid and bile tolerance. The selected 200 LAB isolates with exhibited well-tolerance in acid and bile conditions were then identified by sequencing the 16S rDNA gene, followed by acid and bile tolerance, antimicrobial activity, adhesion to epithelial cells and additional characteristics on the removal of cholesterol. Then, the two probiotic strains (L. ingluviei and L. salivarious) which showed the greatest advantage in vitro testing were selected to assess their efficacy in broiler chickens. Results: It was found that 200 LAB isolates that complied with all measurement criteria belonged to five strains, including L. acidophilus (63 colonies), L. ingluviei (2 colonies), L. reuteri (58 colonies), L. salivarius (72 colonies), and L. saerimneri (5 colonies). We found that the L. ingluviei and L. salivarius can increase the population of LAB and Bifidobacterium spp. while reducing Enterobacteria spp. and Escherichia coli in the cecal content of chickens. Additionally, increased concentrations of valeric acid and short chain fatty acids were also observed. Conclusion: This study indicates that all five Lactobacillus strains isolated from gut contents of chickens are safe and possess probiotic properties, especially L. ingluviei and L. salivarius. Future studies should evaluate the potential for growth improvement in broilers.

First Reliable Record of the Dusky Shark Carcharhinus obscurus (Lesueur, 1818) (Carcharhinidae, Pisces) from Korea (한국산 흉상어과(Carcharhinidae) 어류 1미기록종, Carcharhinus obscurus (Lesueur, 1818))

  • Lee, Woo Jun;Kim, Yoonji;Na, Jung-Hyun;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.6
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    • pp.696-701
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    • 2019
  • Five specimens (839 - 1280 mm TL, Total length) of the dusky shark Carcharhinus obscurus, belonging to the family Carcharhinidae, were collected from the waters off Jejudo Island, Sinan, Namhae, Busan and Gangneung, Korea, during 2010-2019. Carcharhinus obscurus was similar to Carcharhinus brachyurus but was distinguished by the interdorsal ridge (present in C. obscurus vs. absent in C. brachyurus) and the shape of upper jaw teeth (broad in C. obscurus vs. narrow in C. brachyurus). In addition, 479 base-pair sequences in the mitochondrial DNA cytochrome c oxidase subunit I (COI) of our two specimens corresponded to those of C. obscurus (genetic distance, 0.000-0.003), but clearly distinguished from those of the Korean C. brachyurus (genetic distance, 0.03). We adopted the Korean name "Heuk-sang-eo" for Carcharhinus obscurus, after Kim and Ryu (2017).

Identification of Differentially Expressed Genes by TCDD in Human Bronchial Cells: Toxicogenomic Markers for Dioxin Exposure

  • Park, Chung-Mu;Jin, Kyong-Suk;Lee, Yong-Woo
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.1-9
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    • 2012
  • Differentially expressed genes by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were identified in order to evaluate them as dioxin-sensitive markers and crucial signaling molecules to understand dioxin-induced toxic mechanisms in human bronchial cells. Gene expression profiling was analyzed by cDNA microarray and ten genes were selected for further study. They were cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1), S100 calcium binding protein A8 (calgranulin A), S100 calcium binding protein A9 (calgranulin B), aldehyde dehydrogenase 1 family, member A3 (ALDH6) and peroxiredoxin 5 (PRDX5) in up-regulated group. Among them, CYP1B1 was used as a hallmark for dioxin and sharply increased by TCDD exposure. Down-regulated genes were IK cytokine, interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), nuclease sensitive element binding protein 1 (NSEP1), protein tyrosine phosphatase type VI A, member 1 (PTP4A1), ras oncogene family 32 (RAB32). Although up-regulated 4 genes in microarray were coincided with northern hybridization, down-regulated 5 genes showed U-shaped expression pattern which is sharply decreased at lower doses and gradually increased at higher doses. These results introduce some of TCDD-responsive genes can be sensitive markers against TCDD exposure and used as signaling cues to understand toxicity initiated by TCDD inhalation in pulmonary tissues.

What is Epigenetics? -Focusing on Basic Concepts and Mechanisms- (최근 보건의료분야에서 활발하게 연구되고 있는 "Epigenetics"란 무엇인가? -기본개념 및 기전을 중심으로-)

  • Lee, Sun-Dong;Park, Sung-Kyun;Ko, Seong-Gyu;Shin, Heon-Tae;Kim, Myung-Dong
    • Journal of Society of Preventive Korean Medicine
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    • v.14 no.2
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    • pp.1-12
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    • 2010
  • The individual differences in disease development and susceptibility have been researched primarily on the subject of genes, environment or the interaction between genes and the environment respectively. However, there have been limitations in explaining complex diseases, and the differences in health and diseases in monozygotic and dizygotic twins. Fortunately, thanks to active research on the relationship between genes and the environment, and epigenetics, there has been much progress in the understanding of body's reactions and changes. Epigenetics is referred to as a study of gene expression through the interactions of DNA methylation, chromatin's histone and the change of structure in tail, RNA editing without any change in DNA sequence. In this paper, we introduce the basic concepts and mechanisms of epigenetics. The result of the epigenetics is heritable ; can regulate gene expressions ; is reversible ; and has many variable forms depending on cell types. The influences of epigenetics occur throughout life, but it is mainly determined in utero during early pregnancies. Diseases occur or the risk rises if these influences continue after birth until adult life when problems occur in excess/lack of nutrition, environmental plasticity, or already inputted data. Therefore, there is a need for change and innovation, especially in interest and investment in health education for young women near pregnancies and correct treatment of epigenetic-related diseases.

A Probabilistic Approach to the Assessment of Phylogenetic Conservation in Mammalian Hox Gene Clusters

  • Stojanovic, Nikola;Dewar, Ken
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.118-123
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    • 2005
  • With the increasing availability of mammalian genome sequences it became possible to use large scale phylogenetic analysis in order to locate potentially functional regions. In this paper we describe a new probabilistic method for the characterization of phylogenetic conservation in mammalian DNA sequences. We have used this method for the analysis of Hox gene clusters, based on the alignment of 6 species, and we constructed a map of for indicating short and long conserved fragments and their positions with respect to the known locations of Hox genes and other elements, sometimes showing surprising layouts.

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Advances towards Controlling Meiotic Recombination for Plant Breeding

  • Choi, Kyuha
    • Molecules and Cells
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    • v.40 no.11
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    • pp.814-822
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    • 2017
  • Meiotic homologous recombination generates new combinations of preexisting genetic variation and is a crucial process in plant breeding. Within the last decade, our understanding of plant meiotic recombination and genome diversity has advanced considerably. Innovation in DNA sequencing technology has led to the exploration of high-resolution genetic and epigenetic information in plant genomes, which has helped to accelerate plant breeding practices via high-throughput genotyping, and linkage and association mapping. In addition, great advances toward understanding the genetic and epigenetic control mechanisms of meiotic recombination have enabled the expansion of breeding programs and the unlocking of genetic diversity that can be used for crop improvement. This review highlights the recent literature on plant meiotic recombination and discusses the translation of this knowledge to the manipulation of meiotic recombination frequency and location with regards to crop plant breeding.

Combined Effect of CO2 andTemperature on Wheat Powdery Mildew Development

  • Matic, Slavica;Cucu, Maria Alexandra;Garibaldi, Angelo;Gullino, Maria Lodovica
    • The Plant Pathology Journal
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    • v.34 no.4
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    • pp.316-326
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    • 2018
  • The effect of simulated climate changes by applying different temperatures and $CO_2$ levels was investigated in the Blumeria graminis f. sp. tritici/wheat pathosystem. Healthy and inoculated plants were exposed in single phytotrons to six $CO_2$+temperature combinations: (1) 450 ppm $CO_2/18-22^{\circ}C$ (ambient $CO_2$ and low temperature), (2) 850 ppm $CO_2/18-22^{\circ}C$ (elevated $CO_2$ and low temperature), (3) 450 ppm $CO_2/22-26^{\circ}C$ (ambient $CO_2$ and medium temperature), (4) 850 ppm $CO_2/22-26^{\circ}C$ (elevated $CO_2$ and medium temperature), (5) 450 ppm $CO_2/26-30^{\circ}C$ (ambient $CO_2$ and high temperature), and (6) 850 ppm $CO_2/26-30^{\circ}C$ (elevated $CO_2$ and high temperature). Powdery mildew disease index, fungal DNA quantity, plant death incidence, plant expression of pathogenesis-related (PR) genes, plant growth parameters, carbohydrate and chlorophyll content were evaluated. Both $CO_2$ and temperature, and their interaction significantly influenced powdery mildew development. The most advantageous conditions for the progress of powdery mildew on wheat were low temperature and ambient $CO_2$. High temperatures inhibited pathogen growth independent of $CO_2$ conditions, and no typical powdery mildew symptoms were observed. Elevated $CO_2$ did not stimulate powdery mildew development, but was detrimental for plant vitality. Similar abundance of three PR transcripts was found, and the level of their expression was different between six phytotron conditions. Real time PCR quantification of Bgt was in line with the disease index results, but this technique succeeded to detect the pathogen also in asymptomatic plants. Overall, future global warming scenarios may limit the development of powdery mildew on wheat in Mediterranean area, unless the pathogen will adapt to higher temperatures.