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.
The purpose of this study was to evaluate the effect of burs on microleakage of Class V resin restorations when a self-etching primer adhesive was used. Forty Class V cavities were prepared with four different cutting burs on extracted third molars, and divided into one of four equal groups (n = 10); Group 1-plain cut carbide bur (no. 245), Group 2-cross cut carbide bur (no. 557), Group 3-fine diamond bur (TF-21F), Group 4-standard diamond bur (EX-41). The occlusal and gingival margin of cavities was located in enamel and dentin, respectively. Cavities were treated with Clearfil SE Bond and restored with Clearfil AP-X. Specimens were thermocycled, immersed in a 2% methylene blue solution for 24 hours, and bisected longitudinally. They were observed leakages at enamel and dentinal margins. Data were analyzed using Mann-Whitney and Wilcoxon signed ranked test. The results of this study were as follows; 1. At enamel margin, microleakage of group 4 was statistically higher than those of group 1, 2 and 3 (p < 0.01). 2. At dentinal margin, microleakage of group 4 was statistically higher than group 3 (p < 0.01), but group 1 and 2 were not statistically different with group 3 and 4. 3. Enamel microleakage was statistically higher than dentinal microleakage in group 1, 2 and 3 (p < 0.05), but statistical difference between the microleakage of enamel and dentinal margin was not in Group 4. In conclusion, the use of coarse diamond bur showed high microleakage at both enamel and dentinal margin when Clearfil SE Bond was used in class V cavity.
This study was conducted to investigate the developmental ability of interspecies cloned embryos after nuclear transfer of goat fetal fibroblast cells into porcien oocytes. Recipient porcine and goat oocytes were obtained from slaughterhouse and matured in vitro according to established protocols. Enucleation was accomplished by aspirating the first polar body and cytoplasm and a single donor cell was individually microinjected into vitelline space of the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol fusion medium. After electro-fusion, interspecies reconstituted embryos were cultured in PZM-3 for 7 days. In porcine interspecies nuclear transfer with goat fetal fibroblast cells, the cleavage rate of reconstituted embryos were 58.9% which was no significant different from that in porcine nuclear transfer embryos (67.4%). However, the developmental rate into blastocyst stage was 5.4% in interspecies nuclear transfer which was significantly lower than that in porcine intraspecies nuclear transfer (13.6%). When the developmental ability of porcine interspecies nuclear transfer with goat cells was compared with goat intraspecies nuclear transfer, the cleavage rate of embryos were 59.2% and the developmental rate into morular and blastocyst stage was 13.6% in interspecies nuclear transfer which were significantly lower than those in intraspecies nuclear transfer embryos. This result indicated that porcine interspecies nuclear transfer with goat fetal fibroblast cells showed the developmental potential in vitro with lower cleavage and developmental rate compared with intraspecies nuclear transfer.
Journal of the korean academy of Pediatric Dentistry
/
v.26
no.2
/
pp.365-376
/
1999
The importance of finishing and polishing the restoration has been described by several authors. The final step provides for improved metallurgical properties, better marginal adaptation, reduced plaque accumulation. Unfortunately, finishing of the restorations can produce damage from temperature rises at the pulpal wall. The aim of this study was to determine the changes in temperature can be occurred during the use of finishing and polishing instruments under a variety of conditions. ; with or without a water coolant, intermittent or continuous operation, high or low rotation speed, remaining dentin thickness and various restorative materials. Class V preparations were cut on extracted molars and restored with composite resin(Z 100), resin-modified glass ionomer cements(Dyract, Fuji II LC), and amalgam. Finishing was done with aluminum oxide coated disc($Sof-lex^{(R)}$ polishing disc, 3M, USA). The following results were obtained. 1. The rise of temperature during polishing of amalgam restorations was the highest among the all experimental groups except polishing with water coolant(P<0.05). However, there were no statistical differences in temperature rises between Z 100, Dyract and Fuji II LC(P>0.05). 2. The intrapulpal temperature was greatly influenced by the applied time, and intermittent polishing was showed significantly lower temperature rises than continuous polishing(P<0.01). 3. The intrapulpal temperature was increased according to the application of polishing regard less of using water coolant. However, polishing with water coolant showed significantly lower temperature in the pulp than not used water coolant(P<0.01).
This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.
To evaluate the primary production and nutrient uptake of macrophytes in Lake Paldang, this study investigate the vegetation areas of six dominant aquatic plants including Typha angustifolia, Zizania latifolia, Phragmites australis, Trapa japonica, Nelumbo nucifera and Savinia natans, and contents of carbon, nitrogen and phosphorus of each macrophyte. Total vegetation area of six dominant aquatic plants was 1.37 $km^2$. Among them, Typha angustifolia was the most wide-distributed species which occupied the 46.7% of total vegetation area. Littoral zone of South Han river had the largest vegetation area with 0.458 $km^2$, and North Han river, Kyungan river and confluence area in the order named. The results of the contents of carbon, nitrogen and phosphorus of macrophytes showed that the carbon contents of emergent macrophytes was higher than that of other life-forms. The nitrogen content of Salvinia natans, free-floating macrophyte was highest and that of Typha angustifolia, emergent macrophyte was lowest. The phosphorus content of Trapa japonica showed the highest content of phosphorus among six macrophytes and emergent macrophytes such as Zizania latifolia and Phragmites australis showed lower contents of phosphorus than other life-forms. The annual net primary production of macrophytes in Lake Paldang, 2004, was calculated as 758.4 ton C $yr^{-1}$ and the annual net nitrogen and phosphorus uptake of macrophyte was 16,921 kg $yr^{-1}$ and 1,841.0 kg P $yr^{-1}$ respectively. Comparing the total budget of organic carbon, nitrogen and phosphorus in Lake Paldang, the amount of primary production and nutrient uptake by macrophytes take a small portion in total budget implying macrophytes do not play an important role in budget of matters in river-type lake, Lake Paldang.
Park, Sun-Jae;Choi, Seong-Mo;Park, Jong-Seok;An, Kwang-Guk
Korean Journal of Ecology and Environment
/
v.39
no.1
s.115
/
pp.21-31
/
2006
The objectives of this study were to analyze trends of temporal water quality and trophic state in Andong and Imha reservoirs using chemical dataset during 1993 ${\sim}$ 2004, obtained from the Ministry of Environment, Korea. According to long-term limnological analyses, Suspended solids (SS) in Imha Reservoir were 2 ${\sim}$ 8 fold2 greater, than those in SS of Andong Reservoir, and the high solids increased total phosphorus (TP) and biological oxygen demand ($BOD_5$) and decreased the transparency, measured as Secchi depth (SD). Chlorophyll-a (CHL-a) increased little or decreased slightly in the both reservoirs during the high solids, resulting in reduced yields of CHL-a : TP ratios. The deviation analysis of Trophic State Index (TSI) in Imha Reservoir showed that about 70% of TSI (CHL-a)-TSI (SD) and TSI (CHL-a)-TSI(TP) values were less than zero and the lowest values were-60, indicating that influence of inorganic solids (or non-volatile solids) on phytoplankton growth was evident in Imha Reservoir and the impact was greater than that of Andong Reservoir. Inorganic solids in Imha Reservoir resulted in light limitation on phytoplankton growth and thus contributed variations in the relations among three parameters of trophic state index. Especially, seasonal data analysis of nutrients in both reservoirs showed that during the postmonsoon, mean TP concentration was Imha Reservoir greater in than that in Andong Reservoir. The higher TP concentrantion was mainly attributed to increases of inorganic solids from soil erosions and nonpoint source inputs within the watershed. The high inorganic turbidity in Imha Reservoir should be reduced for the conservation of water quality for, especially a tap water supply.
Seo, Han Kyung;Kim, Jeong Ho;Shim, Cheol Min;Kim, Byung Cheol;Choi, Do Cheol;Gwon, Yong Ju;Park, Yung Sun;Kim, Dong Yun
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.
This study assesses of efficiency of oocyte recovery and in vitro development for during the non breeding season in goat. Thirty-four matured goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes were activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I+II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytes. The clavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes. There was no significant difference in oocyte quality between seasonal treatments.
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