• 한국수정란이식학회지
• /
• 제9권2호
• /
• pp.153-160
• /
• 1994

This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.

• 한국의류학회지
• /
• 제31권8호
• /
• pp.1297-1309
• /
• 2007

The purpose of this study is to investigate the effect of shirt color, tie color, tone, perceiver' gender on impression formation by applying tone-in-tone coloration of shirt and necktie: coloration of two colors. This experiment was designed to utilize the $2{\times}6{\times}4{\times}2$ factorial designs. and they are shirt color(red, blue), tie color(red, yellow red, yellow, green, blue, purple), tone(vivid, light, dull, dark), and perceiver' gender(a male, a female). The experimental materials developed for this study were a set of stimulus and response scales. The stimuli were 48 upper body photographs which were color outputs by CAD system(4D-box program). The subjects of this research were 288 male and 288 female college students. There were established impression dimension from factor analysis 27 bi-polar adjectives and investigated the effect of interaction between each variable affected impression formations. The items of the adjectives are classified into 4 Impression dimensions: activity, potency, attractiveness, and tenderness. Shirt color, tie color, tone, and perceiver' gender influenced on impression formation. Specially, tie color and tone influenced on the 4 impression dimension greatly by interaction as well as independently. As a result tie is very important clue of impression formation in tone-in-tone coloration of shirt and necktie. Even though tie occupies only a small area in men's clothes it has great effect since it is close to the face.

• 한국가축번식학회지
• /
• 제16권3호
• /
• pp.239-246
• /
• 1992

미성숙 나포란을 20% FCS가 첨가된 TCM-199으로써 24시간 동안 5% CO2, 8% O2의 공기 조건하에서 체외성숙시킨 다음 난자의 위란강내로 정자를 미세주입하여 체외수정을 실시하였다. 미세주입 하기전 정액을 0.75% BSA가 첨가된 Ham's F-10 배양액으로 2시간 동안 5% CO2, 8% O2의 공기 조건하에서 배양함으로써 수정능 획득을 하도록하였으며 이어 30분 동안 12mM의 dbcGMP와 10mM의 imidazol이 함유된 Ham's F-10 배양액으로 배양함으로써 첨체반응을 유도하였다. 본 실험에서 정자의 미세주입 후 제 2극체와 전핵형성이 일어난 난자의 비율은 각각 9.5, 5.4%였으며 2세포기 이후로 발달한 난자의 비율은 4.1%이었다.

• Clinical and Experimental Reproductive Medicine
• /
• 제23권3호
• /
• pp.357-366
• /
• 1996

The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

• Ocean and Polar Research
• /
• 제32권3호
• /
• pp.229-236
• /
• 2010

A study was conducted to investigate the effect of water temperature and photoperiod on the oxygen consumption of the fasting juvenile Pacific cod Gadus macrocephalus (mean body weight 79.9${\pm}$2.0 g) in order to quantify metabolic response of the species under given conditions. The oxygen consumption rate (OCR) of G. macrocephalus was measured under a combination of four different water temperatures (7, 10, 13 and $16^{\circ}C$) and three different photoperiods (24L:0D, 12L:12D and 0L:24D) with an interval of 5 minutes over a 24-hour period using a closed recirculating respirometer. Three replicates were set up in each treatment. OCRs increased with increased water temperatures under all photoperiod conditions (P<0.001). Mean OCRs at 7, 10, 13 and 16oC ranged from 793.7~1108.4, 1145.7~1570.3, 1352.8~1742.5 and 1458.2~1818.6 mg $O_2$ $kg^{-1}$ $h^{-1}$, respectively. Under all water temperature conditions except $7^{\circ}C$ (P<0.001), mean OCRs of G. macrocephalus were the highest in continuous light (24L:0D) followed by 12L:12D and 0L:24D photoperiods. Mean OCRs of fish exposed to the 12L:12D photoperiod were significantly higher during the light phase than during the dark phase under all temperature conditions (P<0.001). $Q_{10}$ values ranged from 3.19~5.13 between 7 and $10^{\circ}C$, 1.41~1.74 between 10 and $13^{\circ}C$ and 1.15~1.35 between 13 and $16^{\circ}C$, respectively. Based on overall results, water temperature, photoperiod and their combinations exerted a significant influence on the metabolic rate of juvenile cod. This study provides empirical data for estimating the amount of oxygen demand and managing the culture of cod under the given water temperatures and photoperiods.

• Clinical and Experimental Reproductive Medicine
• /
• 제38권3호
• /
• pp.126-134
• /
• 2011

Preimplantation genetic diagnosis (PGD) is gradually widely used in prevention of gene diseases and chromosomal abnormalities. Much improvement has been achieved in biopsy technique and molecular diagnosis. Blastocyst biopsy can increase diagnostic accuracy and reduce allele dropout. It is cost-effective and currently plays an important role. Whole genome amplification permits subsequent individual detection of multiple gene loci and screening all 23 pairs of chromosomes. For PGD of chromosomal translocation, fluorescence $in-situ$ hybridization (FISH) is traditionally used, but with technical difficulty. Array comparative genomic hybridization (CGH) can detect translocation and 23 pairs of chromosomes that may replace FISH. Single nucleotide polymorphisms array with haplotyping can further distinguish between normal chromosomes and balanced translocation. PGD may shorten time to conceive and reduce miscarriage for patients with chromosomal translocation. PGD has a potential value for mitochondrial diseases. Preimplantation genetic haplotyping has been applied for unknown mutation sites of single gene disease. Preimplantation genetic screening (PGS) using limited FISH probes in the cleavage-stage embryo did not increase live birth rates for patients with advanced maternal age, unexplained recurrent abortions, and repeated implantation failure. Polar body and blastocyst biopsy may circumvent the problem of mosaicism. PGS using blastocyst biopsy and array CGH is encouraging and merit further studies. Cryopreservation of biopsied blastocysts instead of fresh transfer permits sufficient time for transportation and genetic analysis. Cryopreservation of embryos may avoid ovarian hyperstimulation syndrome and possible suboptimal endometrium.

• 한국의상디자인학회지
• /
• 제10권2호
• /
• pp.39-56
• /
• 2008

It is a remarkable phenomenon that tattoos has been rapidly spreading among women of modern years. The extremism of feminine ornaments, and pricking and piercing of the skin seem to have deep connection with women's consciousness changing to being independent, active and aggressive. The objective of this study is to analyze the potential quality of today's women, who appears emblematically through tattoo, and to search out the whole aspect and keynote of female tattoos. The methods of research are by examining the general characteristics including concept and history, considering the type and significance of ethnic female tattoos, and then comparing with the symbolism of today's female tattoos. The sphere is limited to women's tattoos from ancient times to today's modern lift, examined from large portions of Africa, Asia, Australasia, the Americas, and Arctic regions. As a result, the following had been reflected in female tattoos in the past. First, incantation, tribe, adult age, adornment, marital status, domestic ability and sex distinction, where among others, the incantatory, ornamental symbol, and domestic sign were representatives. Second, the most popular body parts for having a tattoo were around the lips, hands and abdominal region. Third, the more harsh the environment becomes such as tropical forests, isolated islands, and polar regions, the more number of females have tatoos. Fourth, women's tattoos were much less glamorous and smaller compared to those of men, distinguishing one's sex. On the other hand, today's female tattoos can be classified as five symbols: masculinity, independence, voluptuousness, affection, and purity. These symbols contain women's mentality to pass the limit, and to overcome social inferiority. In modern society, where women must compete with met it has become necessary for women to educe their potential masculine qualities. Being violent accessories for them, tattoos became a mark of such potential. Therefore, in the future, women's interest and demand for tattoos are expected to rise in proportion to the social demand of women's force.

• 한국수정란이식학회지
• /
• 제12권1호
• /
• pp.1-10
• /
• 1997

The present study was conducted to investigate the influence of embryo cell stage at 18h post-fusion and oocyte preactivation on sebsequent in vitro developmental potential in the nuclear transplant rabbit embryos. The embryos of 16-cell stage were collected and synchronized to G$_1$ phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome rnass from the oocytes collected by non-dis-ruptive microsurgery procedure. The separated G$_1$ phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation at 18h post-hCG injection and the separated G$_1$ phase blastomeres of 32-cell stage were injected. Mter culture until 20h post-hOG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The fused nuclear transplant embryos were classified into 3~4-cell, 2-cell and 1-cell stage at 18 hrs post-fusion and cultured until the embryos reached blastocyst stage. The developmental rate to blastocyst stage was significantly (P <0.05) higher in all the reconstituted embryos of 3~4-cell stage(58.0%) than in 2 and icell stage. The developmental rate to blastocyst stage in the embryos of 3~4-cell stage at 18 hrs post-fusion was significantly (P<0.05) higher in the reconstituted without oocyte preactivation(77.8%) than in the oocyte-preactivated embryos (33.3%). These results indicated that the higher rate of in the in vitro development to blastocyst stage might be obtained form the embryos which were reconstituted with nuclear donor of G$_1$ phase and non-preactivated oocyte, and developed more rapidly for 18 hrs post-fusion.

• 한국발생생물학회지:발생과생식
• /
• 제21권4호
• /
• pp.425-434
• /
• 2017

Polyploidy is occurred by the process of endomitosis or cell fusion and usually represent terminally differentiated stage. Their effects on the developmental process were mainly investigated in the amphibian and fishes, and only observed in some rodents as mammalian model. Recently, we have established tetraploidy somatic cell nuclear transfer-derived human embryonic stem cells (SCNT-hESCs) and examined whether it could be available as a research model for the polyploidy cells existed in the human tissues. Two tetraploid hESC lines were artificially acquired by reintroduction of remained 1st polar body during the establishment of SCNT-hESC using MII oocytes obtained from female donors and dermal fibroblasts (DFB) from a 35-year-old adult male. These tetraploid SCNT-hESC lines (CHA-NT1 and CHA-NT3) were identified by the cytogenetic genotyping (91, XXXY,-6, t[2:6] / 92,XXXY,-12,+20) and have shown of indefinite proliferation, but slow speed when compared to euploid SCNT-hESCs. Using the eight Short Tendem Repeat (STR) markers, it was confirmed that both CHA-NT1 and CHA-NT3 lines contain both nuclear and oocyte donor genotypes. These hESCs expressed pluripotency markers and their embryoid bodies (EB) also expressed markers of the three embryonic germ layers and formed teratoma after transplantation into immune deficient mice. This study showed that tetraploidy does not affect the activities of proliferation and differentiation in SCNT-hESC. Therefore, tetraploid hESC lines established after SCNT procedure could be differentiated into various types of cells and could be an useful model for the study of the polyploidy cells in the tissues.

• 한국임상수의학회지
• /
• 제32권6호
• /
• pp.536-539
• /
• 2015

한살령의 잡종 암캐에서 체내 성숙된 난자를 회수하기 위한 실험이 진행되었다. 혈청 프로게스테론 농도를 이용하여 배란 후 약 72시간 뒤에 개복술을 시행하였고, 비정상적으로 확장된 왼쪽 자궁각이 발견되었다. 양쪽 난소는 각각 여덟 개의 황체를 가지고 있었고, 배지를 난관에 관류시켜서 총 16개의 난자를 회수하였다. 모든 난자는 극체가 돌출되어 있었고, 25 um 미만의 난황주위 공간과 제 2중기의 핵을 가지고 있었다. 본 증례는 자궁수종을 가진 암캐에서 체내 성숙된 난자를 확인한 최초의 연구이며, 생식기 관련 질병을 가진 개과 동물에서 회수된 난자가 보조 생식 기술에 사용될 수 있음을 의미한다.