• Journal of Embryo Transfer
• /
• v.27 no.1
• /
• pp.57-62
• /
• 2012

Efficient oocyte activation is a key step for the success of nuclear transfer in cloning. Ionomycin sequentially combined with 6-DMAP is now widely used to activate normal oocytes for analytical studies of oocyte activation and to activate reconstructed oocytes after nuclear transfer. The present study investigated sources of oocytes, duration of ionomycin and 6-DMAP, laser and electric stimulation in goat oocyte activation in order to optimize the protocols. Goat ovaries were collected in individual abattoirs during the breeding season and were delivered to the laboratory within 6 h in saline with 100 IU/ml streptomycin and 0.05 mg/ml penicillin. The oocytes were denuded from the cumulus cell by pipetting with 0.2% hyaluronidase in PBS at 20~22 hr post maturation. Oocytes with the polar body were selected and assigned to four groups for parthenogenetic activation. To examine the effect of duration of ionomycin treatment, oocytes after 20~22 hr of maturation were treated with 2.5 uM ionomycin for 1 or 5 min times and then cultured in 2 mM 6-DMAP for 2 or 4 hr. The activated oocytes were cultured in mSOF at $38.5^{\circ}C$ in $CO_2$ 5%, $O_2$ 5% and $N_2$ 90% multi incubator. Cleavage and blastocyst development was observed at 48 hr and day 8 of culture $in$ $vitro$, respectively. Activation rates of oocytes exposed to ionomycin for 1 min(86.4%) were significantly higher than those treated for 5 min(74.3%) duration. This indicated that 1 min ionomycin treatment was most suitable for activation of goat oocytes. The duration of 6-DMAP treat duration was in 2 mM 6-DMAP for 2 hr after 1 min exposure to 2.5 uM ionomycin. The activation rate of oocytes incubated in 6-DMAP for 2 hour(82.5%) was significantly higher than those in oocytes treated with 4 hr(75.5%).

• Journal of the korean academy of Pediatric Dentistry
• /
• v.24 no.2
• /
• pp.460-474
• /
• 1997

The purpose of this study was to evaluate and compare the effectiveness of various low-viscosity resin systems used as rebonding agents to prevent microleakage at the margins of class I composite resin restorations. Seventy sound human premolars were selected for experiment. Class I cavities were prepared and each cavity was conditioned with a 37% phosphoric acid for 15 sec, rinsed with water for 15 sec, and dried with compressed air. Bonding agent(Scotchbond Multipurpose, 3M Co.) was applied and a hybrid composite resin (Z-100, 3M Co.) was placed using an incremental technic. The excess cured composite resin was carefully removed with Sof-Lex discs(3M Co.) to expose the original margins of the cavity. The following seven groups were established : group 1 was not rebonded and used as control group ; group 2 was rebonded with a Scotchbond Multipurpose(3M Co.) and finished ; group 3 was rebonded with a Fortify(BISCO) and finished ; group 4 was rebonded with a Concise white sealant(3M Co.) and finished ; group 5 was rebonded with a Concise white sealant(3M Co.) and not finished ; group 6 was rebonded with a P&F sealant(BISCO) and finished; group 7 was rebonded with a P&F sealant(BISCO) and not finished. The specimens were then subjected to 500 thermocycles between 5 & 65 with a 10 see dwell time and immersed in 2% methylene blue dye solution for 24 hours and sectioned with low-speed diamond cutter into two part under water condition. The extent of microleakage at rebonded margins was evaluated microscopically and scored for dye penetration according to the following scale : 0=no dye penetration ; 1=dye penetration to half-way along axial wall between enamel surface and DEJ ; 2=dye penetration beyond halfway along axial wall between enamel surface and DEJ ; 3=dye penetration to the full depth of DEJ or beyond DEJ. Selected samples were prepared for SEM observation to determine the depth of penetration of the rebonding agent into the marginal interface. The obtained results were as follows: 1. In the group 2 and 3, which is rebonded with a Scotchbond Multipupose and Fortify, dye penetration score were decreased significantly than that of group 1 (P<0.05), but group 4 and 6 were not statistically different from group 1(P>0.05). 2. There were significant differences between group 4, 6 and group 5, 7 when compared by dye penetration score (P<0.05). 3. In the SEM observation, Scotchbond Multipurpose and Fortify were penetrated within $30-40{\mu}m$ depth of the outermost surface. However, both sealants were failed to penetrate into the debonded interface.

• 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.

• The Journal of Korean Academy of Prosthodontics
• /
• v.35 no.1
• /
• pp.31-42
• /
• 1997

This study was performed to investigate the surface roughness of the Cerec Vita Mark II polished by various polishing techniques, compare with that of the Vintage enamel porcelain glazed by high temperature glazing technique. All of the Cerec specimen were finished with sequential use of high speed diamond burs(grit 45, 30 and $15{\mu}m$). The groups were divided into 5 groups : Group I : Cerec Vita Mark II block specimens polished with Sof-lex discs. Group II : Cerce Vita Mark II block specimens polished with Two Striper MPS. Group III : Cerce Vita Mark II block specimens polished with Enhance. Group IV : Cerce Vita Mark II block specimens polished with Porcelain laminate polishing FG kit. Group V : Vintage enamel porcelain glazed by high temperature glazing Technique. Each group was consisted of 10 specimens. The surfaces produced were examined quantitatively using a laser specular reflectance machine(Perthen RM600-s, Feinpruf Perthen GmbH., Germany) and qualitatively under SEM(JSM-5400, JEOL, Japan). The Results were as follows : 1. The arithmetic mean roughness value(Ra) in groups 1, 2, 3 and 4 was higher than that of group5. There was statistically significant difference(P<0.05). 2. The arithmetic mean roughness value(Ra) decreased in the following orders : group 1, group 2, group 4, group 3 and there was no statistically significant difference between group 1 and group 2, group 3, and group 4. There was statistically significant difference among group 1, 2, and group 3, 4 and group 5(P<0.05). 3. The maximum individual peak-to-valley-height(Rmax) decreased in the following orders : group 2, group 1, group 4, group 3, group 5 and there was no statistically significant difference between group 1 and 2, group 1 and group 4, group 3 and group 5. There was statistically significant difference among group 1, 2, and group 1, 4 and group 3, 5(P<0.05). 4. The treated surfaces of group 5 had smoother surface than that of groups 1, 2, 3, 4 with SEM.

• Journal of Animal Reproduction and Biotechnology
• /
• v.35 no.2
• /
• pp.142-148
• /
• 2020

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode's albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.

• Reproductive and Developmental Biology
• /
• v.30 no.4
• /
• pp.293-299
• /
• 2006

In order to generate transgenic goats expressing human growth hormone (hGH) in their mammary glands, goat ${\beta}-Casein/hGH$ hybrid gene was introduced into goat zygotes by pronuclear microinjection. DNA-injected embryos were transferred to the oviduct of recipients at 2-cell stage or to the uterus at morula/blastocyst stage after cultivation in glutathione-supplemented mSOF medium in vitro. Pregnancy and survival rate were not significantly different between 2-cell embryos and morula/blastocysts transferred to oviduct and uterus, respectively. One transgenic female goat was generated from 153 embryos survived from DNA injection. Southern blot analysis revealed that the transgenic goat harbored single-copy transgene with a partial deletion in its sequences. Despite of the partial sequence deletion, the transgene was successfully expressed hGH at the level of $72.1{\pm}15.1{\mu}g/ml$ in milk throughout lactation period, suggesting that the sequence deletion had occurred in non-essential part of the transgene for the transgene expression. Unfortunately, however, the transgene was not transmitted to her offspring during three successive breeding seasons. These results demonstrated that goat ${\beta}-casein/hGH$ gene was integrated into the transgenic goat genome in a mosaic fashion with a partial sequence deletion, which could result in a low level expression of hGH and a failure of transgene transmission.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.17 no.2
• /
• pp.48-52
• /
• 2013

Purpose: The Molybdenum which is the raw material of $^{99}Mo-^{99m}Tc$ generator is produced from the nuclear reactor. However, output has dwindled as the two nuclear reactors supplying the bulk of radioactive material-one in Chalk River, Ontario and the other in Petten, the Netherlands-have been closed for repairs or maintenance. This resulted in the enhancement of its price. So $^{99}Mo-^{99m}Tc$ generator using$(n,{\gamma})^{99}Mo$ is developed by Korea Atomic Energy Research Institute (KAERI). Medicinal availability of this generator is evaluated in this study. Materials and Methods: The radioactivity of $^{99m}Tc$ eluted in generator 1, 2 and 3 unit developed by KAERI was measured. The quality control test of generator such as appearance test, pH test, LAL test, sterility test, chemical impurity (Al) test and radiochemical purity test were performed. Planar and SPECT/CT image sof SD rat (6 weeks, Female) at 2 hr after injection of $^{99m}Tc-HDP$ (hydroxymethylenediphosphonate) (TechneScan HDP, Malinckrodt Medical, Dutch) and $^{99m}Tc-DPD$ (diphosphono-1, 2-propanedicarboxylicacid) (TECEOS, CIS bio international, France) which were labeled with $^{99m}Tc$ eluted in KAERI and commercial generator (40.5 GBq, Malinckrodt Medical, Dutch) using SPECT/CT camera (Symbia, Siemense, Germany) were obtained respectively. Results: The mean radioactivity of $^{99m}Tc$ elution generator 1unit was 4.18 GBq (113 mCi), generator 2 unit was 4.73 GBq (128 mCi) and generator 3 unit was 3.33 GBq (90 mCi). All quality control tests were within normal limit except pyrogentest. Pyrogen test was positive. Planar and SPECT/CT images of rat injected $^{99m}Tc-HDP$ which was labeled with $^{99m}Tc$ eluted in commercial generator show increased uptake in bone, stomach and bowl. Planar images show increased uptake in liver and bone in case of $^{99m}Tc-DPD$. However, images of rat injected $^{99m}Tc-HDP$ and $^{99m}Tc-DPD$ which were labelled $^{99m}Tc$ eluted in KAERI generator show increased uptake in bone, liver and spleen. Conclusion: If shortcoming is removed such as pyrogen and liver appearance, domestic role as an alternative generator is thought to be able to fill and to secure the national medical service by supplying $^{99m}Tc$ when the supply of $^{99m}Tc$ be comes short.

• Journal of Chest Surgery
• /
• v.19 no.1
• /
• pp.35-42
• /
• 1986

Prevention of thrombombolism after rosthetic cardiac valve replacement is essential for the patients. About 90% of patients are free of major and minor thromboembolic complications 5 year after replacement of cardiac valves with prosthetic devices when they are under control of anticoagulant therapy. Ticlopidine is a drug that alter platelet function to have an antithrombotic effect. It is an antiaggregating agent which inhibits primary platelet function to have an antithrombotic effect. It is an antiaggregating agent which inhibits primary platelet aggregation induced by ADP and increases the production of prostaglandin $D_{2}$. Aspirin in small doses inhibits platelet synthesis of prostaglandins by irreversibly blocking the enzyme cyclo-oxygenase. Platelet secretion and aggregation are impaired with Ticlopidine and Aspirin. the thromboembolic event sof 54 patient s who were treated with Ticlopidine and Aspirin after cardiac valve replacement were evaluated and compared with that of 79 patients who were treated with Wafarin and Aspirin after the same type of operation. The follow-up period ranged from 4 to 110 months (mean of 48 months). there were 11 major thromboembolic episodes including three deaths in the warfarin goup during mean follow-up period of 56 months. two cases of CVA and one hemoarthrosis were noted due to overdose of Warfarin. Inticlopidine group, there was only one fatal thromboembolic epdisode three month after mitral valve replacement during mean follow-up period of 18 months. Two episodes of hypermenorrhea resulting anemia ere noted in the ticlopidine group. We measured the parameters of platelet function in aggreagation curve of platelet with platelet aggregometer (chrono-log Aggregometer, Model No. 430) Aggregation test was performed with three final concentrations of epinephrine in 10 uM/L, ADP in 5uM/L. 28 patients with prosthetic cardiac valves and 35 healthy volunteers were subgrouped as follows to analyze the effect of antithrombotic drugs used. Group I ; 11 patients treated with 250-500 mg of ticlopidine and 0.5gm of Aspirin as a daily single dose after cardiac valve replacement (14 St. Jude Medical and 1 Carpentier-Edwards, 9 patients with atrial fibrillation among them) Group II ; 10 patients treated with 3-5 mg of Warfarin and 0.75 gm of Aspirin daily to prolong prothrombin time around 20 seconds for more than 6 months and single Aspirin dose was maintained afterward as a life-long regimes(3 St. Jude Medical, 1 Hall-Kaster and 7 Carpentier-Edwards valve, 9 patients in atrial fibrilation). Group III ; 7 patients who quit anticoagulant treatment (Warfarin + Aspirin) 6-12 months after the regime as group II (3 St. Jude Medical. 1 bjork-Shiley, 1 Hall-Kaster, 3 Carpentier-Edwards valve, 2 of them are with atrial fibrillation). Group IV ; 35 healthy vounteers (28 males and 7 females). The following results were obtained. 1. The mean maximal platelet aggregability in Group I induced by 10uM/L epinephrine was 15.6%, and 17.5 and 18.7% in BM in proportion to the induction by 5 and 10 uM/L ADP. 2. The mean maximal platelet aggregability in Group II induced by 10uM/L epinephrine was 16.5%, and 27.4 and 44.7% in BM in proportion to the induction by 5 and 10uM/L ADP. 3. The mean maximal platelet aggregability in group III induced by 10uM/L epinephrine was 65%, and 56.5 and 51.8% in BM in proportion to the induction by 5 and 10 uM/L ADP. 4. The mean maximal platelet aggregability in the normal subjects induced by 10 uM/L epinephrine was 64%, and 65 and 69% in Bm inproportion to the induction by 5 and 10 uM/L ADP. 5. Reversible change of platelet aggregation curve induced by 5 and 10uM/L was noted all of the patients in Group I. conclusion : Ticlopidine is an antiaggregating agent which inhibits primary platelet aggregation induced by ADP, and increases the production of prostaglandin $D_{2}$. Ticlopidine and Aspirin produced a significant inhibition of platelet in the presence of ADP and epinephrine in our study. Acccording to our brief experience, 250 mg of ticlopidine and low dose of Aspirin resulted synergistic superior effect to each drug alone in prevention of thromboembolism after prosthetic cardiac valve replacement.

• Korean Journal of Soil Science and Fertilizer
• /
• v.27 no.2
• /
• pp.111-116
• /
• 1994

These studies were carried out to investigate the possibility on the agricultural utilization of lake sediment collected in the Daecheong Lake. Several parameters to estimate the degree of maturity of compost by windrow system were tested during a 50 day periods, and the results were as follows : During the composting of Daecheong Lake sediment, C/N ratio decreased gradually from 18.9 at the beginning to 13.1 at the final, while CEC increased from 33.5me/100g to 62.5me/100g. The temperature of lake sediment pile was the highest as $52^{\circ}C$ at 15th day and then decreased slowly until $30^{\circ}C{\sim}40^{\circ}C$ after 50 days. The results may be suggested that Daecheong Lake sediment compost reaches an acceptable level of stability after about 50 days. In order to know the effect of the Lake sediment and its compost on the growth of tomato(Lycopersicum esculentum M.), pot experiments were performed. The appiication of lake sediment(AS) itself, sediment organic fertilizer(SOF), and NPK mixed with sediment organic fertilizer(NSF) was more effective for tomato plant height, fresh and dry weight, and chlorphyll content than those of control treatment. Also, the lake sediment compost improved the physico-chemical properties of soil such as pH, total carbon, total nitrogen, available phosphorous and CEC.

• Journal of Bio-Environment Control
• /
• v.10 no.2
• /
• pp.111-117
• /
• 2001

This experiment was conducted to elucidate the effects of foliar application of monopotassium phosphate on the acceleration of ind color and fruit quality of Satsuma mandarin cultivated in the plastic greenhouse. The 'a'value of peel chromaticity increased seasonably with the increasing frequency of foliar application of monopotassium phosphate, but became gradually irresponsive as the fruit approached to harvesting time. Glucose content increased with the number of foliar applications so did the fructose content. Content of reducing sugars tended to increase with the number of foliar applications by 0.32 to 0.41%.mL$^{-1}$ juice in treatment sof ive or more applications. Sucrose content increased gradually with the increasing number of foliar applications, but there was no significatn difference among treatments. Total sugar also increased with the increasing number of foliar applications Generally, the soluble solid level is considered to be representative of fruit quality. Sugar content increased with the number of foliar application up to 5 times in which sugar content increased by 0.93$^{\circ}$Bx as compared to the control. The fruit acidity of the treatment plots decreased as compared to that of the control, but there was no significant difference in fruit acidity among foliar application times.