Depigmentation of skin and hair color in the somatic cell cloned pig

Hwang, Kyu-Chan,Cho, Seong-Keun,Lee, Seong-Hoon,Park, Jong-Yi,Kwon, Deug-Nam,Choi, Yun-Jung,Park, Chankyu,Kim, Jae-Hwan,Park, Keun-Kyu,Hwang, Seongsoo,Park, Soo-Bong,Kim, Jin-Hoi Wiley-Liss, Inc. 2009 Developmental dynamics Vol.238 No.7

<P>Previously, we have successfully produced nine cloned piglets using Duroc donor cells. Among these clones, one showed distinct depigmentation of the skin and hair color during puberty. In this study, we selected a clone with depigmentation to investigate the etiology of the anomaly in somatic cell nuclear transfer. We hypothesized that genes related to Waardenburg syndrome (Mitf, Pax-3, Sox-10, Slug, and Kit) are closely associated with the depigmentation of pig, which was derived from somatic cell nuclear transfer (scNT). Total RNA was extracted from the ear tissue of affected and unaffected scNT-derived pigs, and the transcripts encoding Mitf, Pax-3, Sox-10, and Slug, together with the Kit gene, were amplified by reverse transcription-polymerase chain reaction, sequenced, and analyzed. The cDNA sequences from the scNT pig that showed progressive depigmentation did not reveal a mutation in these genes. Although we did not find any mutations in these genes, expression of the genes implicated in Waardenburg syndrome was severely down-regulated in the affected scNT pig when compared with unaffected scNT pigs. This down-regulation of gene expression may result in a previously undescribed phenotype that shows melanocyte instability, leading to progressive loss of pigmentation. Developmental Dynamics 238:1701–1708, 2009. © 2009 Wiley-Liss, Inc.</P>

Ginsenoside Rg1 activates ligand-independent estrogenic effects via rapid estrogen receptor signaling pathway

Quan-Gui Gao,Li-Ping Zhou,Vien Hoi-Yi Lee,Hoi-Yi Chan,Cornelia Wing-Yin Man,Man-Sau Wong 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenoside Rg1 was shown to exert ligand-independent activation of estrogen receptor(ER) via mitogen-activated protein kinaseemediated pathway. Our study aimed to delineate themechanisms by which Rg1 activates the rapid ER signaling pathways. Methods: ER-positive human breast cancer MCF-7 cells and ER-negative human embryonic kidneyHEK293 cells were treated with Rg1 (10 12M, 10 8M), 17ß-estradiol (10 8M), or vehicle. Immunoprecipitationwas conducted to investigate the interactions between signaling protein and ER in MCF-7 cells. To determine the roles of these signaling proteins in the actions of Rg1, small interfering RNA or theirinhibitors were applied. Results: Rg1 rapidly induced ERa translocation to plasma membrane via caveolin-1 and the formation ofsignaling complex involving linker protein (Shc), insulin-like growth factor-I receptor, modulator ofnongenomic activity of ER (MNAR), ERa, and cellular nonreceptor tyrosine kinase (c-Src) in MCF-7 cells. The induction of extracellular signal-regulated protein kinase and mitogen-activated protein kinase kinase(MEK) phosphorylation in MCF-7 cells by Rg1 was suppressed by cotreatment with small interferingRNA against these signaling proteins. The stimulatory effects of Rg1 on MEK phosphorylation in thesecells were suppressed by both PP2 (Src kinase inhibitor) and AG1478 [epidermal growth factor receptor(EGFR) inhibitor]. In addition, Rg1-induced estrogenic activities, EGFR and MEK phosphorylation in MCF-7 cells were abolished by cotreatment with G15 (G protein-coupled estrogen receptor-1 antagonist). Theincrease in intracellular cyclic AMP accumulation, but not Ca mobilization, in MCF-7 cells by Rg1 could beabolished by G15. Conclusion: Ginsenoside Rg1 exerted estrogenic actions by rapidly inducing the formation of ER containingsignalosome in MCF-7 cells. Additionally, Rg1 could activate EGFR and c-Src ER-independentlyand exert estrogenic effects via rapid activation of membrane-associated ER and G protein-coupled estrogenreceptor.

Comparative proteomic analysis associated with term placental insufficiency in cloned pig

Lee, So-Young,Park, Jong-Yi,Choi, Yun-Jung,Cho, Seong-Keun,Ahn, Jong Deok,Kwon, Deug-Nam,Hwang, Kyu-Chan,Kang, Sung-Jo,Paik, Seung-Sam,Seo, Han Geuk,Lee, Hoon Taek,Kim, Jin-Hoi WILEY-VCH 2007 PROTEOMICS -WEINHEIM- Vol.7 No.8

<P>Somatic cell-derived nuclear transfer (scNT) is a method of animal cloning in which the oocyte reprograms a somatic cell nucleus to divide and execute developmental programs. Despite many successes in this field, cloning by scNT remains very inefficient. Unlike other cloned animals, pigs derived by scNT have placentas with severe villous hypoplasia. To obtain a better understanding of the protein networks involved in this phenomenon, we assessed global protein expression profiles in term placentas from scNT-derived and control animals. Proteomic analysis of term placentas from scNT-derived animals identified 43 proteins that were differentially expressed compared to control animals. Among them, 14-3-3 proteins and Annexin V, which are closely involved in the apoptotic signaling pathway, were significantly down- and up-regulated, respectively. Western blot analysis and immunohistochemistry indicated that down-regulation of 14-3-3 proteins in scNT-derived placentas induced apoptosis of cytotrophoblast cells via mitochondria-mediated apoptosis. Taken together, our results suggest that placental insufficiency in scNT-derived placentas may be due to apoptosis, induced in part by the down-regulation of 14-3-3 proteins and up-regulation of Annexin V. They also indicate that proteomic maps represent an important tool for future studies of placental insufficiency and pathology.</P>

Serial cloning of pigs by somatic cell nuclear transfer: Restoration of phenotypic normality during serial cloning

Cho, Seong-Keun,Kim, Jae-Hwan,Park, Jong-Yi,Choi, Yun-Jung,Bang, Jae-Il,Hwang, Kyu-Chan,Cho, Eun-Jeong,Sohn, Sea-Hwan,Uhm, Sang Jun,Koo, Deog-Bon,Lee, Kyung-Kwang,Kim, Teoan,Kim, Jin-Hoi Wiley-Liss,Inc 2007 Developmental dynamics Vol.236 No.12

<P>Somatic cell nuclear transfer (scNT) is a useful way to create cloned animals. However, scNT clones exhibit high levels of phenotypic instability. This instability may be due to epigenetic reprogramming and/or genomic damage in the donor cells. To test this, we produced transgenic pig fibroblasts harboring the truncated human thrombopoietin (hTPO) gene and used them as donor cells in scNT to produce first-generation (G1) cloned piglets. In this study, 2,818 scNT embryos were transferred to 11 recipients and five G1 piglets were obtained. Among them, a clone had a dimorphic facial appearance with severe hypertelorism and a broad prominent nasal bridge. The other clones looked normal. Second-generation (G2) scNT piglets were then produced using ear cells from a G1 piglet that had an abnormal nose phenotype. We reasoned that, if the phenotypic abnormality of the G1 clone was not present in the G2 and third-generation (G3) clones, or was absent in the G2 clones but reappeared in the G3 clones, the phenotypic instability of the G1 clone could be attributed to faulty epigenetic reprogramming rather than to inherent/accidental genomic damage to the donor cells. Blastocyst rates, cell numbers in blastocyst, pregnancy rates, term placenta weight and ponderal index, and birth weight between G1 and G2 clones did not differ, but were significantly (P < 0.05) lower than control age- and sex-matched piglets. Next, we analyzed global methylation changes during development of the preimplantation embryos reconstructed by donor cells used for the production of G1 and G2 clones and could not find any significant differences in the methylation patterns between G1 and G2 clones. Indeed, we failed to detect the phenotypic abnormality in the G2 and G3 clones. Thus, the phenotypic abnormality of the G1 clone is likely to be due to epigenetic dysregulation. Additional observations then suggested that expression of the hTPO gene in the transgenic clones did not appear to be the cause of the phenotypic abnormality in the G1 clones and that the abnormality was acquired by only a few of the G1 clone's cells during its gestational development. Developmental Dynamics 236:3369–3382, 2007. © 2007 Wiley-Liss, Inc.</P>

Oecophyllibacter saccharovorans gen. nov. sp. nov., a bacterial symbiont of the weaver ant Oecophylla smaragdina

Chua Kah-Ooi,See-Too Wah-Seng,Tan Jia-Yi,Song Sze-Looi,Yong Hoi-Sen,Yin Wai-Fong,Chan Kok-Gan 한국미생물학회 2020 The journal of microbiology Vol.58 No.12

In this study, bacterial strains Ha5T, Ta1, and Jb2 were isolated from different colonies of weaver ant Oecophylla smaragdina. They were identified as bacterial symbionts of the ant belonging to family Acetobacteraceae and were distinguished as different strains based on distinctive random-amplified polymorphic DNA (RAPD) fingerprints. Cells of these bacterial strains were Gram-negative, rod-shaped, aerobic, non-motile, catalase-positive and oxidase-negative. They were able to grow at 15–37°C (optimum, 28–30°C) and in the presence of 0–1.5% (w/v) NaCl (optimum 0%). Their predominant cellular fatty acids were C18:1 ω7c, C16:0, C19:0 ω8c cyclo, C14:0, and C16:0 2-OH. Strains Ha5T, Ta1, and Jb2 shared highest 16S rRNA gene sequence similarity (94.56–94.63%) with Neokomagataea tanensis NBRC106556T of family Acetobacteraceae. Both 16S rRNA gene sequence-based phylogenetic analysis and core gene-based phylogenomic analysis placed them in a distinct lineage in family Acetobacteraceae. These bacterial strains shared higher than species level thresholds in multiple overall genome-relatedness indices which indicated that they belonged to the same species. In addition, they did not belong to any of the current taxa of Acetobacteraceae as they had low pairwise average nucleotide identity (< 71%), in silico DNA-DNA hybridization (< 38%) and average amino acid identity (< 67%) values with all the type members of the family. Based on these results, bacterial strains Ha5T, Ta1, and Jb2 represent a novel species of a novel genus in family Acetobacteraceae, for which we propose the name Oecophyllibacter saccharovorans gen. nov. sp. nov., and strain Ha5T as the type strain.

Ginsenoside Rg1 activates ligand-independent estrogenic effects via rapid estrogen receptor signaling pathway

Gao, Quan-Gui,Zhou, Li-Ping,Lee, Vien Hoi-Yi,Chan, Hoi-Yi,Man, Cornelia Wing-Yin,Wong, Man-Sau The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenoside Rg1 was shown to exert ligand-independent activation of estrogen receptor (ER) via mitogen-activated protein kinase-mediated pathway. Our study aimed to delineate the mechanisms by which Rg1 activates the rapid ER signaling pathways. Methods: ER-positive human breast cancer MCF-7 cells and ER-negative human embryonic kidney HEK293 cells were treated with Rg1 ($10^{-12}M$, $10^{-8}M$), $17{\beta}$-estradiol ($10^{-8}M$), or vehicle. Immunoprecipitation was conducted to investigate the interactions between signaling protein and ER in MCF-7 cells. To determine the roles of these signaling proteins in the actions of Rg1, small interfering RNA or their inhibitors were applied. Results: Rg1 rapidly induced $ER{\alpha}$ translocation to plasma membrane via caveolin-1 and the formation of signaling complex involving linker protein (Shc), insulin-like growth factor-I receptor, modulator of nongenomic activity of ER (MNAR), $ER{\alpha}$, and cellular nonreceptor tyrosine kinase (c-Src) in MCF-7 cells. The induction of extracellular signal-regulated protein kinase and mitogen-activated protein kinase kinase (MEK) phosphorylation in MCF-7 cells by Rg1 was suppressed by cotreatment with small interfering RNA against these signaling proteins. The stimulatory effects of Rg1 on MEK phosphorylation in these cells were suppressed by both PP2 (Src kinase inhibitor) and AG1478 [epidermal growth factor receptor (EGFR) inhibitor]. In addition, Rg1-induced estrogenic activities, EGFR and MEK phosphorylation in MCF-7 cells were abolished by cotreatment with G15 (G protein-coupled estrogen receptor-1 antagonist). The increase in intracellular cyclic AMP accumulation, but not Ca mobilization, in MCF-7 cells by Rg1 could be abolished by G15. Conclusion: Ginsenoside Rg1 exerted estrogenic actions by rapidly inducing the formation of ER containing signalosome in MCF-7 cells. Additionally, Rg1 could activate EGFR and c-Src ER-independently and exert estrogenic effects via rapid activation of membrane-associated ER and G protein-coupled estrogen receptor.

Electrical Activation Enhances Pre-Implantation Embryo Development Following Sperm Injection into <i>In Vitro</i> Matured Pig Oocytes

YOO, Jae-Gyu,HUR, Chang-Gi,PARK, Mi-Rung,PARK, Jong-Yi,HWANG, Kyu-Chan,KIM, Jae-Hwan,KIM, Jin-Hoi,CHO, Seong-Keun Japanese Society of Veterinary Science 2012 The Journal of veterinary medical science Vol.74 No.4

<P>The objective of this study was to evaluate the effect of electrical stimulation (EST) on pronuclear formation, chromosomal constitution, and developmental capability among <I>in vitro</I> matured pig oocytes following intracytoplasmic sperm injection (ICSI). After ICSI, the oocytes were randomly distributed and cultured into 3 groups: the EST activated ICSI group, non-activation ICSI group, and <I>in vitro</I> fertilization (IVF) group. The proportion of oocytes in which 2 pronuclei were formed in ICSI groups was significantly higher in the former groups than in the IVF group (96.2 and 93.5 vs. 64.5%, respectively, <I>P</I><0.05). The cleavage rate was significantly higher in EST activated ICSI group (78.6%) than in the IVF and non-activated ICSI groups (51.8 and 46.0%, respectively, <I>P</I><0.05), as was the proportion of oocytes that developed to the blastocyst stage at day 7 (18.9 vs. 11.6 and 9.1%, respectively, <I>P</I><0.05). Diploid blastocysts were observed in 52.4, 63.0, and 65.2% of oocytes in the IVF, activated, and non-activated ICSI groups, respectively. Eight out of 23 gilts (34.8%) were confirmed to be pregnant in activated ICSI groups, but none of these pregnancies were carried to term. These results show that oocyte activation after ICSI is effective in elevating the cleavage rate and blastocyst development, while ensuring normal chromosome composition. Further research is needed to determine the pregnancy maintenance requirements for ICSI-embryos in pigs.</P>

모바일 의료용 앱의 안전성 평가 기준과 시험

박샛별(Set-Byeol Park),김일곤(Il Kon Kim),이병기(Byoung-Kee Yi),김동섭(Dong-Sub Kim),박기정(Ki-Jung Park),허찬회(Chan-Hoi Hur),권태희(Tae-Hee Kwon) 한국정보과학회 2012 정보과학회 컴퓨팅의 실제 논문지 Vol.18 No.1

최근 스마트 기기가 대중화됨에 따라 다양한 어플리케이션이 개발되고 있다. 의료영역에서도 개인건강관리 기능을 갖춘 어플리케이션에서부터 병원에서 의료진이 사용할 수 있도록 개발된 어플리케이션까지 다양한 모바일 의료용 앱이 개발되고 있다. 모바일 의료용 앱이 사용될 때, 환자에게 잘못된 건강 정보를 제공하거나 의료진에게 잘못된 측정 데이터나 본인이 아닌 다른 사람의 정보가 제공되었을 경우, 의료 서비스 품질이 크게 떨어질 뿐만 아니라 환자의 상태에도 악영향을 미칠 수 있다. 이에 의료영역에서 사용되는 모바일 의료용 앱에 대한 안전성 평가 기준을 제시하고 시험 환경을 설계하였다. 또한 상용화된 모바일 의료용 앱에 안전성 평가 기준을 적용하여 본 논문에서 제시하는 모바일 의료용 앱 안전성 평가 기준의 유효성을 검증하였다. With the recent popularity of smart devices, a wide range of mobile medical apps are being developed, from personal health management to applications for healthcare professionals. When a mobile medical app delivers wrongful health information or incorrect measurement data to physicians, it may result in fatal consequences to patients, let alone deteriorating the quality of services. In this paper, we propose a set of safety evaluation criteria for mobile medical apps and describe the design of testing environment. We then verify the effectiveness of the proposed criteria by applying them to some commercially available mobile medical apps.

원격의료시스템의 성능 평가 기준

박샛별(Set-Byeol Park),송준현(Jun-Hyun Song),도형호(Hyoung-Ho Do),윤정배(Jeong-Bae Yun),김일곤(Il-Kon Kim),이병기(Byoung-Kee Yi),차지훈(Ji-Hun Cha),허찬회(Chan-Hoi Hur),박기정(Ki-Jung Park),김동섭(Dong-Sup Kim) 한국정보과학회 2011 한국정보과학회 학술발표논문집 Vol.38 No.2B

최근 통신 기술의 발전과 다양한 바이오 센서 시스템 구축이 가능해 지면서 원격의료가 여러 분야로 확산되고 있다. 고령화 사회에 따른 노인환자와 만성질환자 증가 농어촌 거주자의 의료비 지출 증가는 건강보험 진료비 및 의료비 지출 증가의 원인이 되고 있다. 농어촌 거주자의 의료비 부담을 줄이고 더 나은 의료 서비스를 제공할 수 있는 방법으로 원격의료는 가장 적합한 대안이 될 수 있다. 정보통신 기술의 발전을 기반으로 농어촌과 같은 의료 취약지를 중심으로 원격의료를 확산하기 위해서는 성능과 안전성이 보장된 원격의료시스템을 공급하는 것이 중요하다. 본 논문에서는 원격의료시스템의 성능을 평가하기 위한 기준 및 시험 방법을 제시하고 평가기준에 대한 유효성을 검증한다.