• Journal of Technology Innovation
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• v.24 no.4
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• pp.277-307
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• 2016

Since 2011, DCF(Discounted Cash Flow) method has been used initiatively for valuating R&D technology assets in the agricultural food industry and recently technology valuation based on royalties comparison among technology transfer transactions has been also carried out in parallel when evaluating the technology assets such as new seed development technologies. Since the DCF method which has been known until now has many input variables to be estimated, sophisticated estimation has been demanded at the time of technology valuation. In addition, considering more similar trading cases when applying sales transaction comparison or industry norm method based on information of technology transfer royalty, it is an important issue that should be taken into account in the same way in the Agri-Food industry. The main input variables used for technology valuation in the Agri-Food industry are life cycle of technology asset, the financial information related to the Agri-Food industry, discount rate, and technology contribution rate. The latest infrastructure building and data updating related to technology valuation has been carried out on a regular basis in the evaluation organization of the Agri-Food segment. This study verifies the key variables that give the most important impact on the results for the existing technology valuation in the Agri-Food industry and clarifies the difference between the existing valuation result and the outcome by referring the support information that is derived through the latest input information applied in DCF method. In addition, while presenting the scheme to complement fragment information which the latest input data just influence result of technology valuation, we tried to perform comparative analysis between the existing valuation results and the evaluated outcome after the latest of reference data for making a decision the input values to be estimated in DCF. To perform these analyzes, it was first selected the representative cases evaluated past in the Agri-Food industry, applied a sensitivity analysis for input variables based on these selected cases, and then executed a simulation analysis utilizing the key input variables derived from sensitivity analysis. The results of this study is to provide the information which there are the need for modernization of the data related to the input variables that are utilized during valuating technology assets in the Agri-Food sector and for building the infrastructure of the key input variables in DCF. Therefore it is expected to provide more fruitful information about the results of valuation.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.27-32
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• 2007

This study was conducted to examine the effects of prostaglandin $F_2{\alpha}(PGF_2{\alpha})$ and prostaglandin $E_2 (PGE_2)$ on the expansion and hatching of bovine embryos. During the in vitro culture, embryos were cultured with the following groups: (1) 0, 1, 10 and 100ng/ml $PGF_2{\alpha}$ (2) 0, 1, 10 and 100 ng/ml $PGF_2{\alpha}$, (3) low concentration of $PGF_2{\alpha}$ ; low concentration of $PGF_2{\alpha}$, (1ng/ml : 1ng/ml), (4) low concentration of $PGF_2{\alpha}$ : high concentration of $PGF_2{\alpha}$ (1ng/ml : 10ng/ml) (5) high concentration of $PGF_2{\alpha}$ : low concentration of $PGE_2$ (10ng/ml 1ng/ml) (6) high concentration of $PGF_2{\alpha}$ : high concentration of $PGE_2$(10 ng/ml : 10ng/ml). In the results of this study, treatment of $PGF_2{\alpha}$ or $PGE_2$ did not affect in vitro development to blastocysts. However, the hatching rates of embryos cultured with 10ng/ml $PGE_2$(10.3%) and 1ng/ml $PGF_2{\alpha}$ 10ng/ml $PGE_2$(22.2%) were significantly (P<0.05) higher than in control (4.3% and 12.7%) and other treatment groups. All groups treated with high concentrations of $PGF_2{\alpha}$ showed decreased hatching rates. Thus, this results suggested that $PGF_2{\alpha}\;and\;PGE_2$ were concerned with the hatching in bovine embryos, and their effects on hatching were different by the concentrations.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.317-322
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• 2005

This study was conducted to improve the efficiency of embryo recovery and to establish the protocols of superovulation in Holstein cows. Sixteen Holstein cows were used the test the efficacy of three superovulation regimens using Folltropin. In the case of regimen 1, CIDR plus with E2 capsule was inserted in cows at the random stage of estrous cycle and the total of 400 mg Folltropin V was adminstered twice a day for 4 days(Folltropin V group). In regimen 2, CIDR was inserted and 3.0 mg estradiol benzoate was administered i.m. next day and the total of 400 mg Folltropin was adminstered twice a day for 4 days(Folltropin V+EB group). For regimen 3, CIDR insertion was same as in the regimen 2 and the total of 400 mg Folltropin diluted with $10\%$ PEG 8,000 was administered once(Folttropin V+PEG 8,000 group). In all the regimens, CIDR were removed on 12th day and 45 mg dinoprost was administered i.m. simultaneously. The heat detected donors were administered 200 ug LH-RH and inseminated twice with 2 straws of frozen semen 12 hours apart. Embryo were collected using Foley catherter in each uterine homs on 6${\~}$8 days after inseminations. The evaluation of collected embryos were according to the IETS manual. The CL responses according to the superovulation treatments were 5.8, 20.6, 24.0 in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively and there were significant different among the treatments(p<0.01). Transferable embyos collected were 3.6$\pm$2.4, 3.3$\pm$l.8 and 2.8$\pm$2.3, in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively. Degenerated and unfertilized embryos in regimen 2 and 3 than regimen 1. These results indicates that superovulation treatments with both multiple injections and a single injection using PEG of Folltropin combined with CIDR insertion at the random stage of estrus cycle can be used to produce Holstein embryos.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.303-309
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• 2005

This study was conducted to examine the effects of OA on metaphase of meiosis II and the mitochondrial activity of cytoplasm in bovine cumulus oocytes complexes(COCs) during in vitro maturation. Hanwoo COCs were collected from the slaughterhouse cow ovaries and matured in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA for the maturation rate of OA concentration. For the maturation effects between OA and cycloheximide(CX), COCs were matured in TCM199 with 25 ug/mL CX, 25 ug/mL CX (6 hrs culture) plus 2 uM OA or 2 uM OA only at a atmosphere $5\%\;CO_2,\;95\%$ air $39^{\circ}C$ for 6, 12, 24 hrs. To evaluate the nuclear types of matured COCs, cumulus cells were removedby $0.5\%$ hyaluronidase sol. and oocytes were fixed in 1:3 acetic acid ethyl alcohol for 30 sec. and then stained with $0.1\%$ basic Fuchsin sol. For the detection of fluoriscent intensity (FI) of matures oocytes, cumulus cells were removed same as performed above and were stained with 20 nM mite tracker for 20 min. at $39^{\circ}C$. Mitochondrial activity of FI in matured oocytes was imaged by laser conforcal microscopy (Fluoview, Olympus, Japan) and were measured scanned face on 5 um from median to endpoint of oocytes. Statical analysis of nuclear types observed the three replicates was carried out with ANOVA and Fisher's protected least significant difference test using the STATVIEW program. FI of matures oocytes was compared the multiples of the least intensity among the measured oocytes. Maturing in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA, metaphase B were showed 72.0, 50.0, 70.0, $68.8\%$, respectively and there were different significant(p<0.05). In the case of treatment with OA and CX, metaphase were $73.8\%,\;8.2\%,\;45.5\%,\;73.7\%$ in $0.1\%$ PVA-TCM199, 25 ug/mL CX, 25 ug/mL CX plus OA or 2uM OA only, respeclively. FI was revealed the increasing tendency during the process of maturation. Whereas FI in CX was decreased about 3 times compared to the other treatments of 6 hrs maturation. We conclude that OA regulates bovine COCs maturation and induces the mitochondrial activity during the process of maturation.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.35-43
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• 2006

Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.157-162
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• 2006

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.163-168
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• 2006

The present study was carried out to investigate in vitro development and post-thawed survivability of bovine embryos according to different ovary transport temperatures. Bovine ovaries were collected at a local slaughterhouse and were transported at 4 different temperature categories to laboratory: $7{\sim}10^{\circ}C\;(T1),\;11{\sim}17^{\circ}C\;(T2),\;18{\sim}25^{\circ}C\;(T3)$ and above $26^{\circ}C$ (control group). The cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured. The rates of maturation (to metaphase II), cleavage and development to blastocysts were compared among treatment groups. Furthermore, frozen-thawed blastocysts were in vitro cultured to compare the survivability among groups. The maturation rates in the T1, T2 and T3 groups ($60.0{\sim}68.2%$) were significantly lower than that in the control group (81.8%, p<0.05). The cleavage rates in the T1 and T2 groups (52.6 and 54.5%) were significantly lower than that in the control group (83.6%, p<0.05). However, there was no difference in the development rate to blastocysts among all groups ($27.9{\sim}33.0%$, p>0.05). The survivability of frozen-thawed embryos was significantly lower in the T1 group (46.2%) than those in the T2, T3 and control groups ($68.8{\sim}7.13%$, p<0.05). In conclusion, the results suggest that ovary transport temperature at $26^{\circ}C$ may be optimal for the better in vitro development and the survival of frozen-thawed embryos produced in vitro Furthermore, exposure of ovary to temperature below $10^{\circ}C$ during transport may significantly decrease both in vitro development and survivability of frozen-thawed blastocysts.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.169-175
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• 2006

The objective of this study was carried out to examine the polar body extrusion of in vitro matured porcine follicular oocytes as a non-invasive marker of oocyte quality to know the developmental competence in advance. The porcine oocytes matured for 48 hours were examined the polar body extrusion and some parts were stained. The examined oocytes were matured for additional $16{\sim}18$ hours and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 hours for diploid formation. The treated oocytes were washed and cultured for 7 days. The polar body extrusion and degeneration rates were varied with $9.9{\sim}52.4%$ and $21.4{\sim}61.8%$ by repetition. The polar body extruded oocytes were shown the polar body chromosome and metaphase II plate by staining. However the non-extruded oocytes were shown expanded nucleus with 39.1%, premature chromosome condensation with 19.6%, metaphase I plate with 10.9 %, metaphase II with 13%, condensed chromatin with 6.5%, and absent nuclear material with 8.7%. The oocytes that were not examined for the polar body extrusion were cleaved 45.0%, and developed to blastocyst stage with 11.3%. In examined oocytes for polar body extrusion,. the polar body extruded oocytes were cleaved 94.2% and developed with 42.5%. This result suggests that discarding of the degenerating oocytes and oocytes that not extruded polar body will be effective for experiments of culturing effect in porcine embryos and embryo biotechnology.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.121-126
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• 2007

The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.177-184
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• 2005

This study was conducted to investigate the effects of recombinant bovine somatotropin (rbST) injection on conception and parturition rates in normal or repeat breeding Hanwoo. We treated 462 cows containing 79 repeat-breeding cows of multiparous and allocating 5 treatment groups. Treatment 1 (T1) was injection of 2ml saline (for pseudo treatment), T2 was one injection of rbST 250mg into the tailhead region at the estrus, T3 was twice injection of rbST 250mg both at the time of insemination and again 10 to 14 day later, T4 was once injection of rbST 500mg at insemination and T5 was twice injection of 500mg rbST both at the time of insemination and again 10 to 14day later respectively. In rbST treated groups, timed artificial inseminations (TAI) were performed fellowing estrus synchronization. 100 us GnRH was injected into the scapula region on Day 0, 25mg $PGF_2{\alpha}$ was injected on Day 7 for degeneration of corpus luteum (CL) and 100ug GnRH was injected for inducing the synchronization. The results are as fellows; When normal Hanwoo were inseminated once with rbST administration, the pregnancy rate of T2 $(67.5\pm18.48\%)$ were higher than control $(52.4\pm9.72\%)$, while the pregnancy rate of T4 $(63.3\pm5.77\%)$ were significantly higher (p.<0.05) than control $(39.3\pm12.89\%)$ in repeat breeder Hanwoo. The parturition rates of normal Hanwoo were no differences among the treatments but were significant different in repeat breeder Hanwoo (p<0.05). When the estrous was induced by Ovsynch and inseminated once with rbST administration, the pregnancy rates of T2 was $12.5\%$ higher than control in normal Hanwoo, T4 $(80.0\%)$ was highest among the treatments (p<0.05) in repeat breeder Hanwoo. When normal Hanwoo were inseminated once with rbST administration, the pregnant period was $282.7\~284.8$ days and the body weight was $25.1\~25.9kg$, there were no difference among the treatments. The ratio of sex was almost same without T4 (male vs. female=18:9). In repeat breeder Hanwoo, pregnant period was 280.4~289.3 day and body weight was $23.0\~26.6kg$, it had no difference among the treatments. The sex ratio were similar to normal Hanwoo except T4 (M : F=2 : 8). In conclusion, the pregnancy and parturition rate by once insemination could be improved by the administration of rbST 250mg in normal Hanwoo or 500mg in repeat breeder Hawoo.