• Journal of Animal Reproduction and Biotechnology
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• v.34 no.3
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• pp.205-211
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• 2019

In this study, we investigated the possibility of using mouse embryonic stem cell conditioned medium (ESCM) and embryonic stem cell medium (ESM) for in vitro maturation in the efficient in vitro production of blastocysts from porcine follicular oocyte. Depending on the concentration of supplement of ESCM added to the NCSU-23 solution did not affect 2-cell development rates and blastocysts development. However, in particular, the survival rate (10 days of culture) of blastocyst was significantly higher than that of the control group as the additive concentration (30%) increased (p < 0.05). The survival rate of blastocysts showed a similar tendency even with addition of ESM (30%) alone. On the other hand, the duration of the addition of these additives during IVM (0-44 h) was that the IVM I period (0-22 h) were more effective than the IVM II period (22-44 h). Thus, the effect of these additives is probably due to the combination of the various physiologically active substances of ESCM or the appropriate amino acids and vitamins of ESM. In particular, these additives were more effective during the first half (IVM I) of in vitro maturation. In summary, optimization of ESCM or ESM supplementation may improve in vitro maturation of porcine oocyte and affect developmental competency. Therefore, if more efficient methods of adding ESCM or ESM to basal culture medium can be developed during in vitro maturation of porcine follicle oocytes, high quality blastocysts will be developed from low porcine follicular oocyte compared to other domestic animals.

• Reproductive and Developmental Biology
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• v.38 no.4
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• pp.165-170
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• 2014

Cows may suffer impaired ovarian function, often accompanied by reduced conception rates and increased embryonic loss. Cystic ovarian disease (COD) is one of the most frequently diagnosed gynecological findings in dairy cattle. It causes temporary infertility and is likely to affect reproduction as well as production parameters in cattle. Therefore, the purpose of this study was to determine the expression patterns of apoptosis (Bcl-2, Bax), implantation (E-cadherin) and immune related proteins (TNF-${\alpha}$, IL-10) in uterine endometrium of Hanwoo (Korean native cattle) with ovarian cyst and normal ovarian follicles. In the Western blot analysis, the expression of anti-apoptotic Bcl-2 protein was significantly higher in endometrium with normal ovarian follicles, whereas expression of pro-apoptotic Bax protein was significantly lower. Also, the expressions of E-cadherin and TNF-${\alpha}$ proteins were significantly higher in uterine endometrium with normal ovarian follicles. On the other hand, the expression of IL-10 protein was significantly lower in uterine endometrium with normal ovarian follicles. Taken together, our results provided that the expressions of apoptosis, adhesion and immune related proteins in uterine endometrium with ovarian cyst were showed the aberrant patterns, and we suggest that different expression changes of these proteins may be affect to pregnancy ability of cattle.

• Journal of Embryo Transfer
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• v.8 no.2
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• pp.151-157
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• 1993

The present study was undertaken to determine the optimal condition for parthenogenetic activation of rabbit oocytes by electric stimulation in vitro in an attempt to develop nuclear transplantation techniques for cloning mammalian embryos and animals. Freshly ovulated oocytes were collected from superovulated rabbits from 13 to 26 hrs. after hCG injection. The cumulus-free oocytes were activated parthenogetically by repeated stimuli of square direct electric pulses in O.3M mannitol solution. After applying electric stimulations of different voltages, pulse durations and pulse times, all of the oocytes were cultured in TCM-199 with 10% FCS for 96 hours in a 5% $CO_2$ incubator, and their developmental potential in vitro was examined. The higher activation rate (68.9%) was achieved at the voltage of 2.0kv/cm, the pulse duration of 60 $\mu$sec and three pulse times and the activation rate of 100% was achieved at the pulse duration of 100 and 200 $\mu$sec, the voltage of 1.5kv/cm and three pulse times. Although the higher rates of activation of oocytes were achieved at 100 and 200 $\mu$sec, none of them developed to blastocyst in vitro. The oocytes collected 18~20 hours post hCG injection showed the highest rate of activation and development to blastocyst in vitro than the oocytes collected 13~15 or 25~26 hours post hCG injection. Therefore, it can be suggested that the application of electric stimulation of 2.0kv/cm, 60 $\mu$sec and three pulse times to the oocytes collected at 18~20 hours post hCG injection would be more beneficial for the parthenogenetic activation of oocytes in rabbits.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.203-209
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• 2002

The aim of this study was to assess the developmental capacity of oocytes maturated in vitro after 10 days of culture when the preantral follicles were isolated from juvenile mice 10- and 20-day old, respectively, and to develop in vitro culture system that observed a view to morphology of follicles and nucleus maturation of oocytes. The antral-like cavities became formation after 6 days of culture in follicle isolated from 10- and 20-day old mice. The number of follicles were 21.5 and 33.3 in ovary isolated from 10- and 20-day old mice, respectively. The diameters of oocytes were 51.85 and 57.50 ${\mu}{\textrm}{m}$ before culture and were grew 55.95 and 63.11 ${\mu}{\textrm}{m}$ after culture for 10 days, in follicles isolated from 10- and 20-day old mice, respectively. The observation rates up to the M II and from GV to M II were 4.3 and 22.1%, and 14.5 and 61.1% after culture for 10 days in follicles isolated from 10- and 20-day old mice, respectively.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.18 no.2
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• pp.122-132
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• 2008

The purpose of this study is to present the basic materials of a developmental counterplan by assessing prevalence of musculoskeletal symptoms and characteristics of work-related ergonomic risk factors. The prevalence of musculoskeletal symptoms of 64 vineyard workers was investigated by using a self-reported questionnaire. And the vineyard workers' tasks were also collected and analyzed during 9-month period by observing and videotaping methods to identify the ergonomic risk factors. The prevalence rate of musculoskeletal symptoms of female(84.4%) was much higher than that of male(62.5%) and a total prevalence rate was 73.4%. There was no different prevalence of musculoskeletal symptoms between age over 60 years and under 60 years. The prevalence rates of musculoskeletal complaints for each body part were 48.4%, 42.2%, and 35.9% for the wrist, legs/knee, and shoulder, respectively. In postural risk analysis, 7 works(REBA score) represented the high risk tasks such as delivering harvest boxed(12), picking cluster(11) and so on. The main works(exposure score) were orderly ranked as picking cluster(1590), pruning branch(388), and cluster thinning(327). The risk factors of vineyard work were identified as follows: shoulder flexion(${\geq}45^{\circ}$), wrist Flex./Ext.(${\geq}15^{\circ}$), hand force(power/pinch-grip), and prolonged standing(${\geq}4hr$). The engineering solutions including an improvement of hand tools, working process, and working environment should be applied to the high risky tasks in order to resolve the ergonomic problems. The administrative solutions such as improving a distribution of resting time, an exercise cure, an early recognition of symptoms and rehabilitation might be another solution for reducing musculoskeletal symptoms in vineyard workers.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.139-148
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• 2010

Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

• International Journal of Industrial Entomology and Biomaterials
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• v.24 no.1
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• pp.29-35
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• 2012

Bumblebees are widely used to pollinate various crops, especially tomato, in greenhouses and fields. Here, we investigated whether different sugar solutions have any effects on the oviposition and colony development of Bombus terrestris queens. The types of sugar solution used were a white sugar solution, brown sugar solution, dark brown sugar solution, and prepared sugar solution; all solution contained with a 40% sugar concentration and 0.3% sorbic acid. Among these sugar solutions, B. terrestris queens reared on the white sugar solution exhibited the best results; the rates of oviposition, colony foundation and progeny-queen production were $75.0{\pm}11.0$, $33.0{\pm}9.6$ and $21.7{\pm}7.3$, respectively, which corresponded to 1.1-33.0-fold increases over the results of other sugar solutions. Queens reared on the prepared sugar solution did not found any colonies. The death rate within one month was lowest for the queens reared on the white sugar solution at $26.1{\pm}10.9%$, which was 1.2-2.0-fold lower than the rate for the other sugar solutions. Furthermore, the numbers of progeny produced by queens reared on the white sugar solution, $131.4{\pm}38.8$ workers and $51.0{\pm}40.6$ queens, were also higher, corresponding to 1.1-1.2-fold increases compared to queens raised on the other sugar solutions. Therefore, the white sugar solution was the favorable sugar solution for the egg-laying and colony developmental characteristics of B. terrestris queens.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.207-211
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• 2010

Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.213-216
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• 2010

This study was carried out between November 2008 and October 2009 in the experimental farm of Chunnam National University to investigate the effect of feeding of Sudangrass silage on the reproductive performance of Hanwoo cows. Two diets, rice straw or Sudangrass silage separately from concentrate were fed 19 Hanwoo cows. In control group (=CON), cows (n=9) were fed 5 kg (/head) rice straw and 3 kg (/head) commercial diet. In Sudangrass silage group (=SGS), cows (n=10) were fed 12.5 kg (/head) Sudangrass silage and 1 kg (/head) commercial diet. 1. Days to post-partum insemination were $78.8{\pm}25.3$ days for CON and $84.7{\pm}24.6$ days for SGS group and days to post-partum conception in CON or SGS were $90.1{\pm}24.1$ and $87.7{\pm}26.8$ days, respectively. 2. Post-partum conception rates for first service in CON or SGS was 66.7% and 90.0%, respectively, and caving interval was $375.1{\pm}46.7$ days for CON and $370.2{\pm}34.4$ days for SGS group. 3. Body Condition Scores of Hanwoo cows at artificial insemination was 5.36 for CON group and 4.93 for SGS group.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.253-256
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• 2010

This study was performed to investigate the characteristics within ages and freezing tolerance of spermatozoa in Jindo Dog. Experimental animals were selected 12 herds within 1~8 year's old and collected semen for 2 times in a week. Collected semen was evaluated whole volume and sperm number with CASA system (SIAS, Medical Supply, Korea). Then seminal plasma were separated and diluted with modified Tris-egg yolk extender and added 4, 6 and 8% glycerol for 4 times to final concentration and equilibrated for 1.5 hrs. Before and after freezing, equilibrated semen were evaluated the survival rates. Total volume of sperm at 1~2 year old group is as $5.2{\times}10^8\;cells/ml$ largest and there were no significance among groups. The motility of 1~2 year old group is highest as 90.9% and there were significance among groups. Abnormal sperm showed similar among groups. The survival rate in terms of pre-freezing and post-freezing were decreased all levels of glycerol and reveled 87.0% to 64.5% in 4%, 87.5% to 51.9% in 6% and 73.4% to 29.7% in 8%, there were significant difference among the groups (p<0.05). These results suggest that the optimal sperm-freezing methods in Jindo Dog are utilized with modified Tris egg-yolk extender with 4% glycerol and were improve the reproductive activity by these methods.