• Journal of Embryo Transfer
• /
• v.21 no.1
• /
• pp.35-43
• /
• 2006

Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2007.05a
• /
• pp.39-50
• /
• 2007

Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.

• Journal of Food Hygiene and Safety
• /
• v.32 no.4
• /
• pp.284-289
• /
• 2017

Oxidized polyethylene wax (OPEW) is, one of the food additives, used as a coating agent in citrus fruits and nuts. OPEW is authorized to quantum satis in EU, USA, and is acceptable less than 250 mg/kg in Australia and New Zealand. But OPEW is unauthorized as a food additive in Korea. This study was to establish the analytical method of OPEW and demonstrate the effective application of various food samples. We first conducted to compare the various analytical method including acid value (AV), high temperature gel permeation chromatography (HT-GPC), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS), gas chromatography flame ionization detector (GC-FID) and fourier transform infrared spectroscopy (FT-IR). This result indicated that FT-IR spectrum of OPEW treated food sample displayed absorption bands for carbonyl group (C=O, $1714cm^{-1}$), ester group (C-O, $1463cm^{-1}$), aliphatic group (C-H, $2916cm^{-1}$). Furthermore, IR spectrum of OPEW treated food sample showed similar tendency with IR spectrum of OPEW standard. Therefore, it is confirmed that analytical method using FT-IR can be detected on analysis of OPEW in food. As a result of monitoring of 111 samples using established analytical method, OPEW was not detected in the food samples.

• Journal of Embryo Transfer
• /
• v.20 no.2
• /
• pp.105-112
• /
• 2005

The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

• Journal of Embryo Transfer
• /
• v.21 no.2
• /
• pp.137-146
• /
• 2006

The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

• Journal of Life Science
• /
• v.27 no.11
• /
• pp.1369-1375
• /
• 2017

The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.

• Korean Journal of Food Science and Technology
• /
• v.34 no.2
• /
• pp.255-262
• /
• 2002

This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.

• Journal of Oral Medicine and Pain
• /
• v.38 no.2
• /
• pp.137-142
• /
• 2013

Oral Medicine includes temporomandibular disorder, orofacial pains such as neuropathic pain, soft tissue diseases, halitosis, laser treatment, snoring, sleep apnea and identification through forensic dentistry etc. Such diseases are relatively common and cause great inconvenience and pain to the patients, as well as incur fatal health scare at times. In terms of oral medicine, the number of orofacial patients is growing due to a change in the life style and an increased stress as time goes in contemporary society and the demand of areas requiring oral medical professionalism, such as soft tissue lesions, snoring and sleep apnea, forensic dentistry evaluation and others are rapidly ascending. Consequently, among the areas in dental science, the calls for the expertism in oral medicine and its role are mounting. Analyzing the distribution according to disease entity, symptoms, duration of disease, and the prehistory courses of new patients visiting the department of oral medicine in a year provides information of the role and the relative importance of oral medicine in prospect and enables effective diagnosis and treatments for the patients. Therefore, in the present study, by analyzing new patients visiting the oral medicine clinic in our dental hospital for a year and by evaluating the role and the professionalism in future oral medicine, the authors concluded the followings: 1. It was founded that new patients to oral medicine mainly had temporomandibular disorders, soft tissue diseases, and neuropathic pains. 2. The number of patients with temporomandibular disorder appeared to be the highest percentage and the order within this was the patients with combined disorders, muscle disorder, and internal derangement of joint disc. 3. The number of patients with xerostomia appeared to be the highest percentage within soft tissue disease, followed by lichen planus and recurrent apthous ulcers. 4. The number of patients with burning mouth syndrome appeared to be the highest percentage within neuropathic pain.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.30 no.1
• /
• pp.16-22
• /
• 2014

Purpose: The purpose of this study was to assess the marginal and mesial fitness of zirconia single copings and 3-unit fixed partial dentures (FPDs) manufactured with an identical model. Materials and Methods: An epoxy model in which the maxillary right 2nd premolar is lost and maxillary 1st premolar and 2nd molar are formed as abutments was manufactured and scanned by using a laser scanner. A ten units of zirconia single copings were manufactured for maxillary 1st premolar and 2nd molar, respectively and the same number of 3-unit FPDs were manufactured. For the measurements of fitness, the manufactured silicone replicas were divided into four parts and the fitness were measured by digital microscope at measurement points (P1, P2, P3, P4 and P5) of each plane. The measured gaps were classified into three categories: marginal gap (MG, P1), axial gap (AG, average of P2 and P3), occlusal gap (OG, average of P4 and P5). Results: The ranges of MG, AG and OG for single copings were 18.47 - 40.54 ${\mu}m$, 39.73 - 73.61 ${\mu}m$ and 116.90 - 134.69 ${\mu}m$, respectively. The ranges of MG, AG and OG for 3-unit FPDs were 45.95 - 87.44 ${\mu}m$, 23.78 - 57.00 ${\mu}m$ and 99.89 - 131.06 ${\mu}m$, respectively. Conclusion: The result of the study shows that the MGs for 3-unit FPDs were higher than those of single copings, though they are within the range of clinical acceptance, indicating that the use of more homogeneous zirconia block and modification of sintering processes are needed to ensure the prevention of increase of gap in 3-unit FPDs.

• Journal of the Korean Society of Marine Environment & Safety
• /
• v.24 no.2
• /
• pp.275-281
• /
• 2018

It is known that he pollutant emitted from the combustion process of marine fuel oil causes air pollution and harmful effects to the human body. Accordingly, IMO regulates pollutants emitted from ships. However, the regulation of Particulate Matter (PM) is still in the process of debate, so preemptive action is needed. Fundamental research on PM is essential. In this study, the Dimensionless Light Extinction Constant ($K_e$) of fuel oil used in marine diesel engines was measured and analyzed to construct the basic data of the PM generated from marine-based fuel oil. The fuel oil used in the land diesel engine was measured in the same way for character comparison. Both fuel oils differ in sulfur content and density. The $K_e$ was measured via the optical method using a 633 nm laser and was determined by using the volume fraction of PM collected by the gravimetric filter method. The $K_e$ of the PM discharged from marine fuel oil is 8.28, and the land fuel oil is 8.44. The $K_e$ of two fuel oils was similar within the measurement uncertainty range. However, it was found by comparison with the value obtained by the Rayleigh-Limit solution that the light scattering portion could be large. Also, it was found that light extinction characteristics could be different due to the relationship between light transmittance and collected mass.