• Journal of Bio-Environment Control
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• v.29 no.2
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• pp.130-140
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• 2020

This study was conducted in order to closely examine about optimum shading for superior seedling production a container seedling of Tetradium daniellii, which is being increased the demand for a seedling due to being used for alternative energy, ecological restoration and honey plant. The experiment of investigating the optimum shading on T. daniellii was carried out by using plastic container types (350 ml/cavity) for the forestry facility cultivation. The shading level was treated with full sunlight and with 35%, 55%, 75% of the full sunlight. As a result of having surveyed height and root collar diameter growth of a containerized seedling in T. daniellii, a case of the shading experiment showed a noticeably high value was indicated in the full sunlight. It was surveyed that the stronger shading level leads to the lower growth value. Root development was most active in full sunlight. Dry matter production, it was investigated to be the highest in full sunlight. It was surveyed to be the similar tendency to the outcome of height and root collar diameter growth. QI, which is index of showing the quality of a seedling, stood at 0.98 in full sunlight, thereby having been investigated to be the highest. As for the chlorophyll content in a seedling, the highest chlorophyll content was indicated in the 75% shading treatment with the relatively highest shading level. The photosynthetic rate and the water use efficiency were surveyed to be the highest in full sunlight with 8.48 μmolCO₂·m^-2s^-1, 1.40 μmolCO₂·mmolm^-1H₂O, respectively. As a result of surveying the whole experiment, optimum shading level for superior seedling production a container seedling of T. daniellii is determined in full sunlight (0%). It is expected that this will be used as a basic data for mass production.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.71-77
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• 2001

This experiment was designed to improve the production efficiency of germ-line chimeric mice from phospholipase C (PLC)-$\beta$3 or peroxiredoxin (Prx) E -targeted ($\Delta$) ES cells by the investigating the manipulation conditions and characteristics of Jl ES cells. Four targeting clones were isolated to investigate the karyotypical and morphological stability prior to injection. All clones ($\Delta$PLC$\beta$-3 C3, $\Delta$Prx II C3, C10 and I5) showed more than 80% euploidism, however, most of $\Delta$PLC$\beta$-3 C3 clones were extensively differentiated compared to the other clones. Nine of 13 $\Delta$Prx II chimeras appeared to have at least 80% chimerism, whereas $\Delta$PLC$\beta$-3 C3 chimeras had 20% chimerism at most. Therefore, the morphological stability of ES cells under stable euploidism might mainly affect the production rate of high-coat chimeric mice. To increase the collection rate of injectable blastocysts (IBs), 5 to 10 week -aged C57BL/6J female mice were sacrificed at 3.5 days post-coitum. Ten week-aged mice were the most optimal IB donors by showing the highest collection rate (2.94/mouse) of injectable blastocysts without increase of non-injectable embryos (0.29/mouse). Foster mothers might be another factor because ICR x C57BL/6J F1 foster mother showed more increased productivity in litter size (2.8 vs. 5.6) and chimera (0 vs. 35.3%) than those of ICR foster mothers. In conclusion, the efficient production of germ-line chimeras mainly depends on the maintenance of ES cell morphology during targeting procedure, and the establishment of manipulation conditions might be a key point to maximize it.

• Development and Reproduction
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• v.11 no.2
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• pp.111-119
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• 2007

Differentiation of cells can be induced through modulation of endogenous regulators using exogenous factors. Useful transfection systems to transport a specific exogenous regulator into cell have been tried but still there are many obstacles to overcome. In this study, we examined the transfection efficiency of cell permeable peptides (CPPs) in mouse embryonic stem cell under the various conditions. To identify the CPP-mediated translocation of a protein, we employed recombinant CPP-enhanced green fluorescent protein (EGFP). Viability of R1 cells was different between experimental groups depending on the kind of CPP and the concentration of CPP-EGFP. Translocation of CPP-EGFPs into the R1 cells was not detected until 30 min after CPP-EGFPs treatment in all groups. After 1 hr, translocation of pEP-1-EGFP-N was detected, but it could not be detected in the other group. Transfection of pEP-1EGFP-N was independent on its concentration. The time course did not show saturation even after 24 hr in pEP-1-EGFP-N. These results showed that the permeability depended on the kind of CPP and the location of His-tag in the case of examined CPPs, and did not need biological energy. On summary, the efficiency of transfection of CPP-EGFP depends on the CPP sequences but the culture time is not a key factor in transfection for the mouse embryonic stem cell. For the future studies to improve the efficiency of translocation of protein into embryonic stem cells, it is needed to develop modified CPP or mediator. The studies would be very useful to induce the differentiation of embryonic stem cells.

• Proceedings of the KSAR Conference
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• 2005.06a
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• pp.47-54
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• 2005

In boars, unlike the cases in males of other species, gonadal hormones suppress voluntary feed intake. for this reason, barrows, compared with gilts or boars, eat too much feed resulting in excessive fat deposition. Two experiments were performed in the present study to investigate the effects of implantation of Revalor H[Experiment(Exp.) I: 140mg trenbolone acetate(a synthetic androgen) + 14mg estradiol-$17\beta(E_{2}\beta)$] and Compudose(Exp. II; 24mg $E_{2}\beta$) on growth efficiency, carcass characteristics and circulating concentrations of IGF-I and IGF-binding protein-3(IGFBP-3). In Exp. I, sixty-four cross-bred finishing barrows weighing approximately 60kg were randomly divided into eight pens under a 2[control vs Revalor implant] $\times$ 2(ad libitum vs $80\%$ ad libitum feeding) $\times$2[control($103\%$ NRC-recommended level) vs low-energy($87\%$ NRC recommendation) diet] arrangement of treatments. In Exp. II, effects of Compudose were studied using 80 finishing barrows(10 animals/pen). In both Exps., all the animals were slaughtered at 100- to 110-kg body weight. Both Revalor and Compudose implants caused a decrease in feed intake and backfat thickness without affecting major physicochemical characteristics of the carcass and an increase in circulating IGF-I concentration. Moreover, Revalor implant exhibited greater effects than restricted feeding, low-energy diet, or Compudose in these variables. In addition, Revalor implantation suppressed weight gain, but enhanced the feed efficiency without exhibiting any interaction with the diet or feeding. In summary, results suggest that 1) both androgen and estrogen suppress voluntary feed intake and backfat deposition and enhance IGF-I secretion and 2) these effects of the gonadal steroid hormones in growth are likely to be mediated, in part, by IGF-I in finishing barrows.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.31-36
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• 2005

The purpose of this study was an improvement of efficiency and quality in the production of Korean Native Cow embryos. We investigated effects of concentration of ammonia in in vitro maturation (IVM) medium. In addition, we examined effects of addition or exchange of IVM medium on subsequent development and the cell numbers of blastocysts. The concentrations of ammonia in IVM medium was significantly increased by the increasement of IVM duration (p<0.05). The development rates to the 2 cell-, 8 cell- and blastocyst-stage embryos with the addition of IVM medium were similar among treatment groups. The number of inner cell mass (ICM) cells and the total cell number (TCN) of blastocysts were not differ among treatment groups, whereas the trophectoderm (TE) cell number was significantly lower in the group of 4.5 h addition. The ICM/TCN ratio was significantly higher in the group of 4.5 h addition than in the group of control and 9 h addition. The development rate to the 2-cell embryo with the exchange of IVM medium was significantly higher in the group of 4.5 h exchange and 9 h exchange than in control. The development rate to the blastocyst stage was the highest in the group of 9 h exchange. The number of ICM and ICM/TCN ratio were significantly higher in the group of 9 h exchange than the other groups. The numbers of TE and TCN was similar among treatment groups.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.43-48
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• 2005

To search of a new porcine pheromonal odorants for biostimulation control system technologies to offer a potentially useful and practical way to improve reproductive efficiency in livestock species, the holographic quantitative structure activity relationship (HQSAR) model between odorants, 2-phenoxytetrahydrofurane (A), 2-cyclohexyl-oxytetrahydrofurane (B), derivatives and binding affinity constants (p[Od.]/sub 50/) for porcine odorant-binding protein (pOBP) as receptor of pig pheromones were derivated and disscused. The binding affinity constants of cyclohexyl substituents (A) for pOBP were higher (A>B) than that of phenyl substituents (B). It was revealed that the optimum HQSAR model XI using PLS analyses had a fragment length (5∼8) with chirality at 5 components and hologram length 97 bin, which had a cross-validated q²(predictablities) of 0.916, and a conventional correlation coefficient r² (fitness) of 0.988, respectively. From the atomic contribution, the C3 and C5 atom in 2-oxyfuryl group contributed to binding affinity constants, whereas the central carbon atom in tert-butyl group on the cyclohexyl ring and the C4 atom of furyl group parts showed no contribution.

• Reproductive and Developmental Biology
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• v.31 no.1
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• pp.43-48
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• 2007

This study was conducted to examine the characteristics of fresh and frozen semen, proliferating efficiency by AI, and the coat color of offsprings in Korean Native Striped Cattle (Bos namadicus Falconer, Chikso). Semen were collected from 6 heads of tiger-coated male Chikso. In vitro fertilization (IVF) was conducted with frozen-thawed semen and in vitro matured Korean native brown cattle (general Hanwoo) oocytes. Total 18 heads of Hanwoo and Chikso were inseminated using Chikso semen. Coat colors of total 40 offsprings produced by AI were evaluated. The characteristics of the fresh and frozen-thawed Chikso semen did not differ among individuals. In vitro fertilization rate of Chikso semen was not different from that of general Hanwoo semen. However, developmental rate to the blastocyst stage of IVF embryos was higher in Chikso semen (25.9%) than in general Hanwoo semen (p<0.05). There was no difference in conception rate after AI between Chikso and general Hanwoo. The coat colors of offsprings varied, only 42.5% (17/40 heads) of offsprings had tiger coat color. Futhermore, only 55% of offsprings produced from the tiger-coated recipients had tiger coat color. This result shows that proliferation of Chikso by AI is possible, but further research approaches may be needed to enhance the productivity of tiger-coated Chikso.

• Reproductive and Developmental Biology
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• v.31 no.1
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• pp.1-6
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• 2007

The nonobese diabetic / severe combined immune deficiency (NOD/SCID) has been used for determination of proliferation and differentiation of hematopoietic stem cells as xenotransplantation animal model. In this study, we transplanted porcine hematopoietic cells from bone marrow into NOD/SCID mice via intravenous injection to confirm the activity of differentiation and proliferation for porcine hematopoietic cells in vivo. Interestingly, we observed the result of high efficiency with pig T lymphocytes in hematopoietic organs, liver, spleen lymph node, and bone marrow in NOD/SCID mice. The porcine $CD3^{+}$ T cells were detected with $5.4{\pm}1.9%$ in bone marrow, $15.4{\pm}7.3%$ in spleen, $21.3{\pm}1.4%$ in liver, and $33.5{\pm}32.8%$ in lymph node of NOD/SCID mice at 6 weeks after trans-plantation Furthermore, immunohistochemical analysis showed the high engraftment of porcine T lymphocytes in spleen of NOD/SCID mice. Our data suggest that NOD/SCID mice are excellent animal model to determinate the generation md function of pig T lymphocytes.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.17-22
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• 2011

This study was designed to determine the effect of electric field strength, duration and fusion buffer in fusion parameters on the rate of membrane fusion between the somatic cell and cytoplast for Korean cattle (HanWoo) somatic cell nuclear transfer (SCNT) procedure. Following electrofusion, effect of 5 or $10\;{\mu}M$ $Ca^{2+}$-ionophore of activation treatment on subsequent development was also evaluated. Cell fusion rates were significantly increased from 23.1% at 20 V/mm to 59.7% at 26 V/mm and 52.9% at 27 V/mm (p<0.05). Due to higher cytoplasmic membrane rupture or cellular lysis, overall efficiency was decreased when the strength was increased to 30 V/mm (18.5%) and 40 V/mm (6.3%) and the fusion rate was also decreased when the strength was at 25 V/mm or below. The optimal duration of electric stimulation was significantly higher in $25\;{\mu}s$ than 20 and $30\;{\mu}s$ (18.5% versus 9.3% and 6.3%, respectively, p<0.05). Two nonelectrolyte fusion buffers, Zimmermann's (0.28 M sucrose) and 0.28 M mannitol solution for cell fusion, were used for donor cell and ooplast fusion and the fusion rate was significantly higher in Zimmermann's cell fusion buffer than in 0.28 M mannitol (91.1% versus 48.4%, respectively, p<0.05). The cleavage and blastocyst formation rates of SCNT bovine embryos activated by $5\;{\mu}M$ $Ca^{2+}$-ionophore was significantly higher than the rates of the embryos activated with $10\;{\mu}M$ of $Ca^{2+}$-ionophore (70.0% versus 42.9% and 22.5% versus 14.3%, respectively; p<0.05). This result is the reverse to that of parthenotes which shows significantly higher cleavage and blastocyst rates in $10\;{\mu}M$ $Ca^{2+}$-ionophore than $5\;{\mu}M$ counterpart (65.6% versus 40.3% and 19.5% versus 9.7%, respectively; p<0.05). In conclusion, SCNT couplet fusion by single pulse of 26 V/mm for $25\;{\mu}s$ in Zimmermann's fusion buffer followed by artificial activation with $5\;{\mu}M$ $Ca^{2+}$-ionophore are suggested as optimal fusion and activation methods in Korean cattle SCNT protocol.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.55-60
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• 2011

The objective of this study was carried out to evaluate the efficiency of sperm collection methods on the post-thaw viability of cat semen. The cat semen was collected by artificial virginal (AV) and electronic ejaculate (EE) methods. The composition of semen extender was consisted of Tris-buffer supplemented with 20% egg yolk and 1% P/S antibiotics in Ext I, and more added 8% glycerol, 1.0% Equex STM paste of total volume in Ext II. The collected semen was adjusted the concentration and then diluted in Ext I for optimal concentration. The diluted semen was cooling to $5^{\circ}C$ temperature in refrigerator for at least 2 hrs and then diluted stepwise with Ext II for at least 1 hrs. After an equilibration for 1 hrs, the cooled semen was packaged in 0.5 ml straw and then freezing on the $LN_2$ vapor over 5 cm above from $LN_2$ and then immersed directly in $LN_2$ for cryopreservation. The frozen semen was thawed in $38^{\circ}C$ water for 15 sec and then evaluated the motility, viability, and morphology. Post-thaw semen were calculated the motility by SMI (sperm motility index). The live-dead sperm was evaluated by Eosin-B and morphological evaluation was by Diff-quik kit staining. The post-thaw concentration ($89{\times}10^6$ /ml vs. $128{\times}10^6$ /ml), viability ($22.6{\pm}10.6%$ vs. $37.1{\pm}26.1%$), morphological normality ($27.0{\pm}50.2%$ vs. $45.6{\pm}123.0%$) of EE and AV groups were not significant different, but the post-thaw motility was significant lower in EE than that in AV group ($53.1{\pm}3.6$ vs. $73.6{\pm}5.7$) (p<0.05). In conclusion, semen collection methods did not significant different between EE and AV groups except of post-thaw motility and so both semen collection methods could be applied in feline semen collection methods.