• Fisheries and Aquatic Sciences
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• v.24 no.12
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• pp.437-445
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• 2021

Roughscale sole (Clidoderma asperrimum) is only wild caught because basic reproductive research on this species is lacking and gamete production in an artificial setting has not been successful. Exogenous hormone treatment has been used to induce gonadal maturation and final spermiation in wild-caught individuals. In this study, the effects of an exogenous hormone on spermiation in roughscale sole was investigated by implanting different concentrations of a salmon gonadotrophin-releasing hormone analog (sGnRH; 0, 25, 50, and 100 ㎍/kg body weight) into male fishes. The control group did not produce sperm after 21 days post-implantation, and the duration of spermiation was shorter compared to the other groups. The spermiation period and milt amount differed among the hormone-treated groups according to the hormone concentration used. Milt volumes in the groups treated with 25 and 100 ㎍/kg sGnRH increased compared to the control group, whereas exogenous hormone treatment had no effect on the movable sperm ratio. The spermatocrit was high at the beginning of spermiation in all groups and then tended to decrease gradually over time except in the experimental group treated with 100 ㎍/kg sGnRH. Plasma levels of testosterone, 11-ketotestosterone, 17α, 20β-dihydroxy-4-pregnen-3-one were not significantly affected by the sGnRH treatments. Our results demonstrate that it is possible to prolong the spermiation period and increase milt volume by treating male roughscale soles with an exogenous hormone. In addition, the artificial hormone treatment did not affect sperm motility.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.4
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• pp.300-305
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• 2020

Objective: The feasibility of a gonadotropin-releasing hormone agonist (GnRHa) trigger in normal responders is still a matter of debate. The aim of this study was to compare the number of mature oocytes, the number of good-quality embryos, and the live birth rate in normal responders triggered by GnRHa alone, GnRHa and human chorionic gonadotropin (hCG; a dual trigger), and hCG alone. Methods: A retrospective cohort study was conducted at the infertility clinic of a university hospital. Data from 200 normal responders who underwent controlled ovarian hyperstimulation and intracytoplasmic sperm injection with a GnRH antagonist protocol between January 2016 and January 2017 were reviewed. The first study group consisted of patients with cycles triggered by GnRHa alone. The second study group consisted of patients with cycles triggered by both GnRHa and low-dose hCG (a dual trigger). The control group consisted of patients with cycles triggered by hCG alone. Results: The groups were comparable in terms of demographics and cycle characteristics. The numbers of total oocytes retrieved and metaphase II oocytes were similar between the groups. The total numbers of top-quality embryos were 3.2±2.9 in the GnRHa group, 4.4±3.2 in the dual-trigger group, and 2.9±2.1 in the hCG group (p=0.014). The live birth rates were 21.4%, 30.5%, and 28.2% in those groups, respectively (p=0.126). Conclusion: In normal responders, a dual-trigger approach appears superior to an hCG trigger alone with regard to the number of top-quality embryos produced. However, no clinical benefit was apparent in terms of live birth rates.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.69-69
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• 2002

Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.

• Korean Journal of Food Science and Technology
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• v.40 no.6
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• pp.656-660
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• 2008

Fermented soybean foods are an important component of the Korean diet. Eoyukjang is a type of traditional fermented soybean source. Microbial analysis of eoyukjang was conducted during the fermentation period in this study. Microorganisms isolated from eoyukjang were identified by biochemical tests and 16S rDNA sequencing. 17 different microorganisms, including bacteria, yeast, and fungi were detected in eoyukjang during the fermentation period. Even though Aspergillus participated in the early stage of fermentation of eoyukjang, Bacillus species and Saccharomyces cerevisiae were the major microzymes in eoyukjang throughout the maturation period. Eoyukjang is generally consumed after the boiling of the final sample. Therefore, the final sample of eoyukjang was boiled and analyzed. Our results showed that no vegetative microorganisms survived under the boiling conditions for eoyukjang. Fermented soybean products in the domestic market were also assessed for comparison with the results from eoyukjang. The total cell number of kanjang (soy sauce) samples was between 0 to 42 CFU/mL. The isolated microorganisms were identified as Bacillus species. All Bacillus isolates were not found to harbor the three enterotoxin-producing and emetic toxin-producing genes.

• Fisheries and Aquatic Sciences
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• v.22 no.9
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• pp.20.1-20.8
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• 2019

Background: Despite the favorable geo-climatic potential of Cameroon, the national production of tilapia remains low due to poor tilapia growth reported by fish farmers. One of the underlying reasons is the early female maturation at a very small size and precocious breeding in earthen ponds, resulting in overpopulation which leads to stunted growth and therefore to the production of unmarketable fish size. Studies have shown that dietary supplementation of G. kola enhanced growth in young Clarias gariepinus and Oreochromis niloticus. It was also reported that G. kola inhibited spawning in Tilapia adult females. Therefore, this study sought to assess the effects of Garcinia kola as growth promoter and inhibitor of gonadal development in young Oreochromis niloticus. Methods: A total of 108 juveniles weighing $13.32{\pm}0.62g$ were randomly distributed in 9 hapas of 12 fishes each (9 females and 3 males) and fed for 70 days with three isonitrogenous diets, 40% crude protein with increasing Garcinia kola supplementation levels of 0 (normal diet), 6% and 10% (experimental diets). Physico-chemical parameters of the water (temperature, dissolved oxygen, pH, nitrate, nitrite, ammonia, and transparency) were measured twice a week. Every 14 days, fish were harvested, counted, and weighed. At the end of the experiment, three fish of each sex per replicate were sacrificed and their gonad and liver collected and weighed. Data were statistically analyzed using one-way analysis of variance repeated measure followed by Newman-Keuls multiple tests. Results: The results showed that all physico-chemical parameters of the water were within the recommended values for Tilapia culture. Tilapia fed 6% Garcinia kola supplemented diet displayed higher final body weight in males ($38.60{\pm}3.50g$) and females ($36.77{\pm}3.62g$) compared to those receiving normal diet ($36.23{\pm}1.36g$ and $25.87{\pm}3.32g$; respectively to the final body weight in males and females). The gonadosomatic index and hepatosomatic index indicated no significant variation in males while in females, these were significantly low in the experimental fish compared to control fish. Conclusion: The results of this study demonstrated that supplementation of G. kola seeds in diets of young Tilapia improved growth performance and impaired gonadal development in females.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Horticultural Science & Technology
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• v.32 no.3
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• pp.289-295
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• 2014

This research was conducted to investigate a difference in quality according to control of fruit load and cluster weight when seeded 'Heukboseok' grape was grown in a conventional cultivation system. Clusters per vine were set to harvest 1,500 kg, 1,800 kg, and 2,200 kg grapes per $990m^2$. In p lots t reated t o produce 1,800 kg grapes, clusters were set to 3 grades produce grapes with 350 g, 500 g, and 700 g in cluster weight. Based on color chart (National Institute of Horticultural & Herbal Science, Rural Development Admnistration) which show maturation stage for 'Kyoho' grape, grapes with higher than grade 9 were harvested at 80 and 90 days after full bloom, and the r est of them were harvested a t 100 days after full bloom. The final yield of 'Heukboseok' grape was the lowest in target yield of 1,800 kg with 700 g clusters with 75.5% of target yield, but 92.1%-100.1% of target yield were harvested in other treatments. Among treatments with several fruit setting, 83.3% of grapes harvested in treatment to produce 1,500 kg grapes as a target yield at 90 days after full bloom. Among the treatments with different cluster weights, 93.5% grapes were harvested in the treatment with 350 g in cluster weight. 'Heukboseok' grape showed rapid maturation pattern with no significant difference in either soluble solids content or acidity between 90 days from 80 days after full bloom. Therefore, to prevent flesh softening, it is appropriate to harvest fruit at 90 days after full blooms, and both yield and cluster weight were important factors influencing the quality of the fruits in 'Heukboseok' grape. It is confirmed that coloring and firmness was influenced by the final yield and weight per cluster, respectively. Therefore, the yield should be set as grape of 1,500 kg per $990m^2$ in target yield, and cluster weight should be adjusted to 350 g to produce grape of 1,800 kg with the fruits of high quality. It is considered to be hard to produce grape of high quality with 700 g in cluster weight in 'Heukboseok' grape.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.3
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• pp.285-296
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• 1995

Anatomical studies of sink tissues are required for better understanding the biological plant growth system and energy metabolism. Kinematics of root growth zones of two genotypes of tall fescue (Festuca arundinacea Schreb.) receiving 50 or 200 ppm N were determined. Longitudinal anatomy and cell dynamics of root growth zones were studied and calculated. The root growth zone is organized similarly to the leaf growth zone which has cell division, elongation, and maturation zones, but the root growth zone is only about 3.0 mm long compared to 25 to 30 mm for the leaf growth zone. The root cap extends about 0.4 to 0.5 mm from the apical initial, while the cell elongation zone for both cortical and metaxylem cells extends about 3.3 mm from the apical initial for both genotypes and N levels. Root cap cells elongate from an initial length of about 5$\mu{m}$ long to a final length of about 40$\mu{m}$ before being sloughed. Initial lengths of cortical and metaxylem cells were about 8.5 $\mu{m}$ and 13.0 $\mu{m}$, respectively. Elongation of cortex and metaxylem cell showed sigmoidal curves with final lengths of about 120 $\mu{m}$ for cortex cells and 650 $\mu{m}$ for metaxylem cells. Initial size and final size for both types were not affected by N level, but cell fluxes and cell elongation rates of cortical and metaxylem cells were about double in low N. Cell production rates were about 5 to 6 times higher in cortical cells than in metaxylem cells. Differences in N caused a larger change in cell production rate, duration of cell elongation, and relative cell elongation rate than did the genotypes. These data indicate that N application affects root growth longitudinally by changing cell production rate and elongation rate.

• Journal of Nutrition and Health
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• v.48 no.5
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• pp.381-389
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• 2015

Purpose: The objective of this study was to investigate the effects of a high fructose and fat diet on bone growth and maturation in growing female rats. Methods: Three-week-old female SD rats were randomly assigned to four experimental groups; the control group (CON: fed control diet based on AIN-93G, n = 8); the high-fructose diet group (HFrc: fed control diet with 30% fructose, n = 8); the high-fat diet group (Hfat: fed control diet with 45 kcal% fat, n = 8); and the high-fat diet plus high fructose group (HFrc + HFat: fed diets 45 kcal% fat with 30% fructose, n = 8). Each group was assigned their respective diets for the remaining eight weeks. Bone-related parameters (bone mineral density (BMD) and structural parameters, osteocalcin (OC), deoxypyridinoline (DPD)) and morphologic changes of kidney were analyzed at the end of the experiment. Results: Final body weights and weight gain were higher in the HFat and HFrc + HFat groups and showed higher tendency in the HFrc group compared with those of the CON group (p < 0.05); however, no significant difference in caloric intake was observed among the four experimental groups. The serum OC levels of the HFrc and HFrc + HFat groups were lower than those of the CON and HFat groups (p < 0.05). Urinary levels of DPD did not differ among the experimental groups. BV/TV and Tb.N of trabecular bone were higher in the HFrc + HFat group and showed a higher tendency in the HFrc group than those of the CON and HFat groups (p < 0.05). Tb.Pf of trabecular bone were lower in the HFrc + HFat group than those in the CON and HFat groups (p < 0.05). However, no difference in trabecular BMD was observed among the experimental groups. Cortical bone volume was higher in the HFat and HFrc + HFat groups than in the CON and HFrc groups (p < 0.05). No morphology change in kidney was observed among the experimental groups. Conclusion: Our study suggests that 8 weeks of high-fructose and high fat intake could improve the bone quality (Structural parameters) of trabecular and cortical bone of tibia in growing female rats.

• Journal of Aquaculture
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• v.19 no.4
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• pp.267-274
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• 2006

To understand the changes in plasma levels of sex steroids in the wild Japanese eel Anguilla japonica during artificially maturing process, eels received weekly intraperitoneal injections of a water-in-oil (W/O) type emulsion with Freund`s incomplete adjuvant containing salmon pituitary extract (SPE; 20 mg pituitary powder/fish) were examined. In the weekly Eel's Ringer-treated control wild eels, the body weight (BW) changes of fish decreased slowly during the experiment period. Plasma testosterone (T), $estradiol-17{\beta}\;(E_2)$ and $17a,20{\beta}-dihydroxyprogesterone$ (DHP) levels did not change significantly at the end of the experiment. In the weekly SPE-treated silver eels, however, rapid increase in BW changes occurred after 6 to 10 weeks, and the oocytes of all fish were observed to be in the migratory nucleus stage. Furthermore, significant increase in sex steroid hormones (T and $E_2$) levels occurred from 6 weeks. In the weekly SPE-treated yellow eels, the BW changes of fish increased slowly at 6 weeks and then increased. In these fish, the oocytes were at the tertiary yolk globule stage even at the end of the experiment. Plasma sex steroid hormones profiles revealed individual variability in SPE-treated yellow eels. Plasma T and $E_2$ levels significantly increased at 8 weeks and after 6 weeks, respectively, in SPE-treated yellow eels. In the weekly SPE-treated wild eels (silver and yellow eels), however, plasma DHP levels did not change significantly during the experiment period. In silver eel, final maturation could be induced by weekly administration of SPE using W/O type emulsion.