bFGF-defendent feline induced pluripotent stem cells reprogrammed by Sendai virus vector

Myeong-Don Joo,Ji-Yun Hwang,Kyeong-Lim Lee,Seok-Hwan Song,James Kehler,Il-Keun Kong 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

The production of feline induced pluripotent stem cells (iPSCs) can solve the problems that are related with existing unstable supply and demand of eggs as well as ethical aspects about embryonic stem cell at the same time. On the basis of excellent proliferation, it is to facilitate the researches about human disease like FIV and Allergen at the level of cells, not experimental animals. But, a lot of advanced researches are lean too much towards on the transduction using DNA type virus that have the risk of tumorigenesis during reprogramming and on the mLIF-dependent culture condition for the production of feline iPSCs. This being so, this study shows the reprogramming results using Sendai virus vector that is RNA type virus and have no the footprint after transduction. In addition, the feline iPSCs were stably cultured in bFGF-dependent culture condition during the reprogramming step and culture step. In conclusion, we found the bFGF-dependent culture condition in feline iPSCs and suggested the approach using Sendai virus vector as an alternative for reprogramming without concern about tumorigenesis. These methods can be universally applicable to not only the researches about reconstruction and conservation of feline species, but also to a lot of deep studies related with iPSCs or LIF, bFGF to find new approaches.

한국형 출혈열의 심전도학적 고찰

조명찬,주인종,김대중,이병직,정형배,이명묵,박영배,최윤식,서정돈,이영우 대한내과학회 1990 대한내과학회지 Vol.38 No.3

We reviewed 549 electrocardiograms obtained from 95 patients with Korean hemorrhagic fever to investigate the incidence and electrocardiographic characteristics, observe evolutional changes in those findings during the course of illness, identify the correlations with clinical findings, and then suggest pathophysiologic mechanisms in the development of abnormal electrocardiographic findings. The overall incidence of electrocardiographic abnormalites was 95.7%, and the frequency of each phase was 87.8% in the febrile phase, 91.3% in the hypotensive phase, 86.5% in the oliguric phase, 76.4% in the diuretic phase and 43.0% in the convalescent phase. The cardinal electrocardiographic findings were QT prolongation(82.1%), sinus bradycardia(73.7%), prominent U wave(53.7%), tall and peaked T wave(51.6%), ST-T changes(41.1 %) and sinus tachycardia(35.8%). The most frequently observed electrocardiographic findings of each phase were sinus tachycardia in the febrile and hypotensive phase, QT prolongation in the oliguric phase, sinus bradycardia in the diuretic phase and prominent U wave in the convalescent phase. The occurrence of abnormal electrocardiograms increased in patients with high serum creatinine levels during the oliguric and diuretic phases. The electrocardiographic abnormalities that persisted to the convalescent phase were QT prologation(seven cases), prominent U wave(seven cases), sinus bradycardia(five cases), and tall and peaked T wave(four cases). Even though almost all of the electrocardiographic abnormalities were mild and recovered shortly after the convalescent phase and there were no diagnostic electrocardiograms, we observed noteworthy electrocardiographic characteristics and evolutional changes during the course of illness and suggested the primary electrocardiographic changes in Korean hemorrhagic fever.

Effect of kisspeptin on the proliferation and apoptosis of bovine granulosa cells

Hongyu Liu,Myeong-Don Joo,Ji-Yoon Hwang,Ayman Mesalam,Jun Wang,Wenfa Lu,Il-Keun Kong 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Previous studies have shown that kisspeptin (Kp-10) is expressed in mammalian ovaries; however, the expression and role of Kp-10 in bovine ovarian granulosa cells are still unclear. In this study, we assessed the expression of Kp-10 and its effects on the proliferation and apoptosis of bovine granulosa cells. Immunohistochemical analysis showed that Kp-10 was expressed in the cytoplasm of bovine ovarian granulosa cells. Moreover, MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2- H-tetrazolium bromide) assays showed that 100 nM Kp-10 significantly inhibited the viability of granulosa cells (P<0.01). Flow cytometry analysis showed that Kp-10 could significantly increase accumulation of cells in the G1 phase, decrease accumulation of cells in the S phase, and promote apoptosis in bovine granulosa cells (P<0.05). Additionally, Kp-10 decreased the mRNA levels of Bcl-2, an anti-apoptotic gene; increased the mRNA levels of caspase-3, a pro-apoptotic gene; and increased the mRNA levels of Fas and Fasl, two membrane surface molecule genes (P<0.05). Thus, our findings demonstrated for the first time that Kp-10 inhibited proliferation and promoted apoptosis in bovine ovarian granulosa cells. These findings provide insights into our understanding of the role of Kp-10 in mediating the proliferation of bovine granulosa cells.

Holo-transferrin can increase cleave rates of the bovine embryo after in vitro fertilization

MMR Chowdhury,Myeong-Don Joo,Ayman Mesalsm,Lianguang Xu,Fahmida Afrin,Yeong-Gyu Ko,Sung Soo Lee,Sung Woo Kim,Il-Keun Kong 한국실험동물학회 2019 한국실험동물학회 학술발표대회 논문집 Vol.2019 No.7

Low white-matter integrity between the left thalamus and inferior frontal gyrus in patients with insomnia disorder

Kang, Jae Myeong,Joo, Sung Woo,Son, Young-Don,Kim, Hyun,Ko, Kwang-Pil,Lee, Jung Sun,Kang, Seung-Gul Joule Inc. 2018 Journal of psychiatry & neuroscience Vol.43 No.6

Effect of MMP-2 & -9 on the Improvement of Mouse Pregnancy Potential

Shimin Zhang,Ayman Mesalam,Myeong-don Joo,Ji-Yoon Hwang,Seon-hwa Oh,Muhammad Idress,Hongyu Liu,Il-Keun Kong 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

Matrix metalloproteinases (MMPs) comprise a family of zinc-dependent proteinases. Proteins of the MMPs family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development. Successful implantation is closely linked to the expression of MMPs which greatly influences ability of an embryo to degrade the basement membrane of the uterine epithelium, mainly composed of type IV collagen, and invade the uterine stroma. The purpose of this study was to determine the MMP-2 & -9, also known as type IV collagenase, concentrations in mouse uterine fluid during implantation window, investigate the roles of MMP-2 & -9 on mouse trophoblastic cells invasion in vitro and study the effect of MMP-2 & -9 cotransfer with mouse embryos on reproductive performances. Protein level was detected by Elisa kit and invasiveness was assessed by an invasion assay. Optimal concentrations of MMP-2 and -9 was co-transferred to 2.5 dpc recipients using the non-surgical embryo transfer. Our results showed that MMP-2 & -9 protein levels in mouse uterine fluid were significantly increased at 2.5 dpc. Moreover, in vitro treatment significantly promoted both spreading and invasion of mouse trophoblastic cells as compared with the non-treated. Embryo transfer results showed that MMP-9 cotransfer enhanced mouse implantation and pregnancy rate compared with the control and MMP-2 cotransfered groups, however, that was not significant. Taken together, our findings imply that MMP-9 cotransfer with embryos can regulate embryo invasion during preimplantation, which may have serious consequence on embryo implantation and can be applied in other mammals so far as human being.

A fetal whole ovarian culture model for the generation of complete in vitro fertile oocytes from mouse primordial germ cells

Ji-Yoon Hwang,Myeong-Don Joo,Hongyu Liu,Muhamad Idress,Sun-Hwa Oh,Il-Keun Kong 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Mammalian fetal ovaries contains numerous primordial germ cells, however fewer ones can yield mature oocytes due to apoptosis and follicle atresia. Successful in vitro reconstitution of primordial germ cells has recently had a significant effect in the field of assisted reproductive technologies. However, the regulatory mechanisms underlying oogenesis remain unknown and recapitulation of oogenesis in vitro remains unachieved. Therefore, development of methods for obtaining mature oocytes by culturing the fetal ovaries in vitro could contribute to clarify these mechanisms. We adapt an in vitro system for culturing mouse fetal ovaries that support successful follicle assembly and improve oocyte growth and maturation. Ovarian tissues from 12.5 days postcoitum (dpc) fetal mice were cultured in vitro and the matured oocytes were differentiated from primordial germ cells after a 31 days culture period. Our results demonstrate that mouse fetal germ cells are able to form primordial follicles with artificial ovarian cells, and that oocytes within the growing follicles are able to mature normally in vitro. Taken together, this in vitro culture system is expected to aid in the development of new strategies to identify the reasons behind failure of follicle assembly and offer a platform for innovative research into preservation of female germ cells and conservation of endangered species.

Dynamics of connexin expression in the lycopene treated oocytes of Korean native cow

MMR Chowdhury,Ayman Mesalam,Myeong-Don Joo,Lianguang Xu,Fahmida Afrin,Yeoung-Gyu Ko,Sung Soo Lee,Sung Woo Kim,Il-Keun Kong 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Ovarian folliculogenesis and the production of fertilizable oocytes depend on gap junctional intercellular communication within both the developing and the mature follicle. Gap junctions connect oocytes with granulosa cells and granulosa cells with each other. Various nutritional bio-molecules are known to be transferred to the growing oocyte from the granulosa cells via gap junction. Signals that regulate meiotic maturation of fully-grown oocytes pass through the oocyte-granulosa cell gap junctions. Gap junctions also play a critical role in regulating uterine blood flow, contributing to the maternal recognition and also implantation during pregnancy. Due to the challenge of various stressors the in vitro embryo developmental potentials are still suboptimal compared to in vivo. To identify the molecular mechanism of these stressors and to improve the existing embryo developmental potentials, the singlet oxygens quencher lycopene was added to the culture media to counterbalance the oxidative damage caused by ROS. In this study, we have patterned connexin like Cx43, Cx37, Cx32 and Cx26 at protein and transcription level during follicular growth, atresia and blastocyst stage by using immunohistochemistry, conventional PCR and RT-qPCR. Lycopene (0.2 μM) significantly (P < 0.05) increased the gap junctional communication protein (connexin) expression of Cx43, Cx37, Cx32, Cx26 as compared to the control group at both transcription and translation level during follicular growth, atresia and blastocyst stage. Lycopene potentiates ovarian folliculogenesis, provides the production of fertilizable oocytes and improved embryo developmental capabilities by increasing gap junctional intercellular communication.

Effects of Serum Type in Maturation Medium on In Vitro Development of Bovine Embryos

Ayman Mesalam,Shimin Zhang,Lianguang Xu,Myeong-Don Joo,Ji-Yoon Hwang,Kyeong-Lim Lee,Il-Keun Kong 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

This study investigated the effect of Charcoal-Dextran Stripped fetal bovine serum (CDS FBS) and heat-inactivated FBS (HI FBS) in maturation medium on their ability to support in vitro oocyte maturation, cumulus cell-oocyte gap junctional communication, and development of bovine embryos. Oocytes were cultured in TCM-199 supplemented with either 8% BSA, 10% CDS FBS, or 10% HI FBS and 1 μg/mL estradiol-17β, 10 μg/mL FSH, 10 ng/mL EGF, 0.6 mM cysteine, 0.2 mM sodium pyruvate, and followed by in vitro fertilization and the zygotes were cultured in SOF-BE1 medium. The developmental ability and quality of bovine embryos were determined by assessing their cell number, lipid content, mitochondrial activity, gene expression, immunocytochemistry, and cryo-tolerance. The differences in embryo development between experimental groups were analyzed by one-way ANOVA. We have shown that CDS FBS had a higher significant (p<0.05) effect on the rate of blastocyst formation comparing to HI FBS and BSA (45.2±0.7% vs. 37.4±1.5% and 31.1±3.9%, respectively, six replicates were performed). Culture of oocytes with CDS FBS increased (p<0.05) the expression of gape junction proteins, CX37 and CX43, at both transcriptional and translation levels. We also found that CDS FBS significantly increased total cell number and decreased the apoptotic index in day-8 blastocyst comparing to BSA group. Furthermore, the beneficial effects of CDS FBS on embryos were associated with significantly reduced intracellular lipid content and increased mitochondrial activity in both oocytes and blastocyst. Taken together, these data suggest that supplementation of maturation medium with CDS FBS affect cumulus cell-oocyte gap junctional communication, and subsequently improved in vitro developmental competence of bovine oocytes and embryos.

Takayasu 동맥염의 임상적 고찰

최기준(Kee Joon Choi),조주희(Joo Hee Zo),한경일(Kyung Il Han),김영권(Young Kwon Kim),조명찬(Myeong Chan Cho),손대원(Dae Won Sohn),이명묵(Myoung Mook Lee),박영배(Young Bae Park),최윤식(Yun Shik Choi),서정돈(Jung Don Seo),이영우(Young 대한내과학회 1990 대한내과학회지 Vol.39 No.5

N/A To observe the clinical features of Takayasu's arteritis and evaluate the incidence and sites of coronary arterial involvement in Takayasu's arteritis, clinical observations were made in 99 patients who were diagnosed as Takayasu's arteritis by clinical features and aortographic findings in Seoul National University Hospital from August 1971 to July 1989 Coronary arteriographies were performed prospectively in 30 patients with Takayasu's arteritis since March 1987. The results were as follows: 1) In 99 cases of Takayasu's arteritis, 15 were male and 84 were female. The male to female ratio was 1:5.7, the mean age was 29.5, and 63% of those studied were under the age of 30. 2) The following were the presenting clinical symptoms and signs in decreasing order of frequency: headache(69%), weak or nonpalpable pulse(68%), carotid or abdominal bruit(55%), Dyspnea on exertion(51%), hypertention(48%), and dizziness(43%). 3) Aorta or arterial involvement in decreasing order of frequency: left subclavian artery(62%), abdominal aorta(58%), right subclavian artery(38%), descending thoracic aorta(37%) and left renal artery(35%). 4) Using Ueno's classification, 29(29%) were Type I, 22(22g) were Type II, and 48(48%) were Type III. Type IV or pulmonary arterial involvements were seen in 11(33%) out of 33 patients. 5) In a prospective study using coronary arteriography, 8(27%) out of 30 patients of Takayasu's ateritis showed coronary involvement. Among the 13 lesions of coronary arterial narrowings in 8 patients with coronary involvements, there were 3 ostial lesions, 5 proximal and 5 middle or distal lesions. 6) Frequently there were no cardiac symptom in patients with Takayasu's arteritis who had coronary arterial lesions, and a myocadial infarction or congestive heart failure may be the first sign of the coronary arterial narrowing.