식이 칼슘, 비타민 D의 섭취와 골다공증, 고혈압 및 당뇨병의 상관관계

신현아 ( Hyun Ah Shin ),엄애선 ( Ae Son Om ) 한국유가공기술과학회 2009 Journal of Dairy Science and Biotechnology (JMSB) Vol.27 No.2

In the light of the increasing concern over the prevalence of osteoporosis, hypertension, diabetes mellitus, we performed this study to review the correlation between the dietary calcium and vitamin D intake and these diseases. To this end, we investigated the effects of dietary calcium and vitamin D on these diseases. We observed that the intake of dietary calcium and vitamin D had a negative correlation with the incidences of osteoporosis, hypertension, and diabetes mellitus. Further, the intake of these two nutrients is expected to improve related mechanisms such as the renin-angiotensin system. Therefore, we suggest that dietary calcium and vitamin D have a beneficial effect on these diseases.

단위발생 유래 생쥐 배아줄기세포의 기능성 심근세포 형성

신현아,김은영,이영재,이금실,박은미,이훈택,정길생,박세필,임진호,Shin, Hyun-Ah,Kim, Eun-Young,Lee, Young-Jae,Lee, Keum-Sil,Park, Eun-Mi,Lee, Hoon-Taek,Chung, Kil-Saeng,Park, Se-Pill,Lim, Jin-Ho 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.2

Objective : This study was to establish a reproducible differentiation system from the parthenogenetic mouse embryonic stem (P-mES02) cells into functional cardiomyocytes like as in vitro fertilization mouse embryonic stem (mES01) cells. Materials and Methods: To induce differentiation, P-mES02 cells were dissociated and aggregated in suspension culture environment for embryoid body (EB) formation. For differentiation into cardiomyocytes, day 4 EBs were treated with 0.75% dimethyl sulfoxide (DMSO) for another 4 days (4-/4+) and then were plated onto gelatin-coated dish. Cultured cells were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. This frequency was compared with the results of DMSO not treated P-mES02 group (4-/4-) and mES01 groups (4-/4+ or 4-/4-). For confirm the generation of cardiomyocytes, beating cell masses were treated with trypsin-EDTA, dispersed cells were plated onto glass coverslips and incubated for 48 h. Attached cells were fixed using 4% paraformaldehyde and incubated with specific antibodies (Abs) to detect cardiomyocytes (anti-sarcomeric ? -actinin Ab, 1 : 100; anti-cardiac troponin I Ab, 1 : 2000) for 1 h. And the cells were finally treated with FITC or TRITC labelled 2nd Abs, respectively, then they were examined under fluorescence microscopy. Results: Rhythmically contracting areas in mES01 or P-mES02 cells were firstly appeared at 9 or 10 days after EBs plating, respectively. The highest cumulative frequency of beating EBs was not different in both treatment groups (mES01 and P-mES02, 4-/4+) with the results of 61.3 % at 13 days and 69.8% at 15 days, respectively. Also, the contracting duration of individual beating EBs was different from minimal 7 days to maximal 53 days. However, DMSO not treated groups (mES01 and P-mES02, 4-/4-) also had contracting characteristics although their frequency was a few compared to those of DMSO treated groups (6.0% and 4.0%). Cells recovered from the spontaneously contracting areas within EBs in both treated groups were stained positively with muscle specific anti-sarcomeric ? -actinin Ab and cardiac specific anti-cardiac troponin I Ab. Conclusion: This study demonstrated that the P-mES02 cell-derived cardiomyocytes displayed similarly structural properties to mES01 cell-derived cardiomyocytes and that the DMSO treatment enhanced the cardiomyocytes differentiation in vitro.

Tyrosine Hydroxylase 유전자가 주입된 인간 배아줄기세포의 체외 신경세포 분화

신현아,김은영,이금실,조황윤,김용식,이원돈,박세필,임진호,Shin, Hyun-Ah,Kim, Eun-Young,Lee, Keum-Sil,Cho, Hwang-Yoon,Kim, Yong-Sik,Lee, Won-Don,Park, Se-Pill,Lim, Jin-Ho 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.1

Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

Effect of Fertilization Promoting Peptide (FPP) on the Acrosome Status of Cryopreserved Human Sperm

박세필,신현아,김은영,이원돈,임진호,Park, Se Pill,Shin, Hyun Ah,Kim, Eun Young,Lee, Won Don,Lim, Jin Ho The Korean Society for Reproductive Medicine 2005 Clinical and Experimental Reproductive Medicine Vol.32 No.2

연구목적: 정자의 첨체상태는 수정능과 상관관계가 있다. 본 연구는 사람 정자의 동결보존 시 Fertilization promoting peptide (FPP) 처리가 첨체 유지에 효과가 있는지를 알아보고자 실시하였다. 연구재료 및 방법: 사람 정자는 정액검사를 의뢰한 시료를 사용하였으며, 적정농도를 조사하기 위하여 25, 50, 100 nM FPP를 신선정자에 처리한 뒤 시간별로 첨체의 변화를 조사하였다. 또한 적정화된 50 nM FPP를 정자의 동결-융해 시에 처리한 뒤 첨체 변화를 조사하였다. 첨체 변화는 FITC - pisum sativum lectin (PSA) 염색방법을 이용하여 조사하였다. 결 과: FPP 농도 변화와 처리시간에 따른 사람 정자의 첨체 변화를 조사하였던 바, 50 nM FPP 처리군에서 대조군보다 높은 온전한 첨체비율을 얻을 수 있었다. 정자의 동결-융해 시, 동결액과 융해액에 50 nM FPP 첨가가 온전한 첨체를 유지하는 비율을 조사하였던 바, 신선 정자의 결과보다는 유의하게 낮지만 무 처리군보다 유의적으로 높은 온전한 첨체를 얻을 수 있는 것을 알 수 있었다. 또한 동결액에만 또는 융해액에만 50 nM FPP 처리를 하더라도 무 처리군보다 유의하게 높은 온전한 첨체 비율을 획득할 수 있음을 알 수 있었다 (p<0.001). 결 론: 사람 정자의 동결보존 시 50 nM FPP 첨가는 자발적으로 발생하는 첨체반응을 억제하고, 온전한 첨체를 유지할 수 있어 수정능 보유에 기여할 수 있을 것으로 사료된다.

인간정자의 처리에 있어서 Percoll과 Sil-Select 방법의 비교

문신용,류범용,신현아,오선경,서창석,김석현,최영민,김정구,최규홍,이진용,Moon, Shin-Yong,Ryu, Buom-Yong,Shin, Hyun-Ah,Oh, Sun-Kyung,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Choi, Kyu-Hong,Lee, Jin-Yong 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4

Objective: To evaluate silane-coated silica particles (Sil-select) as an alternative to polyvinylpyrrolidone-coated particles (Percoll) for gradient separation of spermatozoa, for use in assisted reproduction. Methods: 20 normal semen based on WHO criteria were included in this study. Recovery of motile and morphologically normal spermatozoa after using two-layer Percoll and Sil-select gradient respectively was recorded. Motility, HOST (hypoosmotic swelling test) and the detection of malondialdehyde for LPO (lipid peroxidation) after 24 h of incubation at $37^{\circ}C$ in a 5% $CO_2$ incubator were compared. Results: Percoll (78.5%) and Sil-select (79.1%) showed a significant increase in the motility compared to ejaculate (60.9%) but no difference between Percoll and Sil-select. Normal sperm morphology significantly increased after Percoll (57.6%) and Sil-select (53.7%) compared to ejaculate (35.8%) but no difference between Percoll and Sil-select. No differences in the recovery of motile spermatozoa and motility, HOST and the production of malondialdehyde after 24 h incubation were found when comparing the use of Percoll and Sil-select. Conclusion: Sil-select seems to be an attractive alternative to Percoll for sperm separation in assisted reproduction.

신경성장촉진 인자가 인간 배아줄기세포 유래 도파민 분비 신경세포형성에 미치는 영향

이금실,김은영,신현아,조황윤,왕규창,김용식,이훈택,정길생,이원돈,박세필,임진호,Lee, Keum-Sil,Kim, Eun-Young,Shin, Hyun-Ah,Cho, Hwang-Yoon,Wang, Kyu-Chang,Kim, Yong-Sik,Lee, Hoon-Taek,Chung, Kil-Saeng,Lee, Won-Don,Park, Se-Pill,Lim, Jin 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.1

Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

체외수정 유래 생쥐 배아줄기세포와 유사한 특성을 보유한 단위발생 유래 생쥐 배아줄기세포

박세필,김은영,이금실,이영재,신현아,민현정,이훈택,정길생,임진호,Park, Se-Pill,Kim, Eun-Young,Lee, Keum-Si,Lee, Young-Jae,Shin, Hyun-Ah,Min, Hyun-Jung,Lee, Hoon-Taek,Chung, Kil-Saeng,Lim, Jin-Ho 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.2

Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.

공동학술대회 발표논문 : 건강기능식품법의 개정 및 의의

엄애선 ( Ae Son Om ),신현아 ( Hyun Ah Shin ),이상화 ( Sang Hwa Yi ) 한국법정책학회 2009 법과 정책연구 Vol.9 No.1

This study was designed to survey the revision of the Health Functional Food Law and the meaning of it. In the latest revision of the Health Functional Food Law was undertaken in March, 2008. In this law firstly, the form of tablet, capsule, powder, granule, liquid, pill, etc. were deleted so that the range of application for the health functional food could be extended. Secondly, the history-tracking control system for the health functional food was revised in order to improve the safety. Thirdly, the procedures of the complaint motions for health claim and deliberation on advertisement of the health functional food were amended. Lastly, the payment range and the criteria for the compensation were made. Taking all of these into account, it is suggested that the revision of the Health Functional Food Law is reasonable and desirable.

자기제조 배추김치와 시판 김치의 이화학적 품질 실태 조사

이상화 ( Sang Hwa Yi ),박신영 ( Shin Young Park ),정단희 ( Dan Hee Jeong ),김진영 ( Jin Young Kim ),이애정 ( Ae Jung Lee ),신현아 ( Hyun Ah Shin ),문지혜 ( Ji Hea Moon ),이진혁 ( Jin Hyeuk Lee ),김성언 ( Sung Eon Kim ),유현주 ( Hy 한국식품조리과학회 2009 한국식품조리과학회지 Vol.25 No.6

미취학 어린이의 손씻기 교육에 따른 미생물학적 실태 분석

엄애선(Ae Son Om),김지은(Ji Eun Kim),문지혜(Ji Hea Moon),신현아(Hyun Ah Shin),이지선(Ji Sun Lee),권성희(Sung Hee Kwon),이정숙(Jung Sook Lee) 한국조리학회 2011 한국조리학회지 Vol.17 No.3

The objective of this study is to monitor the microbiological effect on preschool children`s hand safety according to educational methods of hand washing. The subjects were ten children aged three to five. The analyses of the microbiological effect were made before hand washing education, after one-week video demonstrations of hand washing education, and after one-week video demonstrations of hand washing education combined with practice. The results were as follows. Total plate count, filamentous fungi, coliform, and Staphylocuccus aureus were detected from children`s hands just before hand washing education. According to the two-week hand washing education, such education was found effective in keeping the children`s hands clean and safe. In conclusion, repeated and more consistent training of hand washing would be important for children, especially aged three to five because most of the habits and behavior patterns were developed in these age groups and the effects were persistent during their future life.