It is highly challenging to measure the efficiency of electric vehicle charging stations (EVCSs) because factors affecting operational characteristics of EVCSs are time-varying in practice. For the efficiency measurement, environmental factors around the EVCSs can be considered because such factors affect charging behaviors of electric vehicle drivers, resulting in variations of accessibility and attractiveness for the EVCSs. Considering dynamics of the factors, this paper examines the technical efficiency of 622 electric vehicle charging stations in Seoul using data envelopment analysis (DEA). The DEA is formulated as a multi-period output-oriented constant return to scale model. Five inputs including floating population, number of nearby EVCSs, average distance of nearby EVCSs, traffic volume and traffic congestion are considered and the charging frequency of EVCSs is used as the output. The result of efficiency measurement shows that not many EVCSs has most of charging demand at certain periods of time, while the others are facing with anemic charging demand. Tobit regression analyses show that the traffic congestion negatively affects the efficiency of EVCSs, while the traffic volume and the number of nearby EVCSs are positive factors improving the efficiency around EVCSs. We draw some notable characteristics of efficient EVCSs by comparing means of the inputs related to the groups classified by K-means clustering algorithm. This analysis presents that efficient EVCSs can be generally characterized with the high number of nearby EVCSs and low level of the traffic congestion.

Evaluating the operational efficiency of electric vehicle charging stations (EVCSs) is important to understand charging network evolution and the charging behavior of electric vehicle users. However, aggregation of efficiency performance metrics poses a significant challenge to practitioners and researchers. In general, the operational efficiency of EVCSs can be measured as a complicated function of various factors with multiple criteria. Such a complex aspect of managing EVCSs becomes one of the challenging issues to measure their operational efficiency. Considering the difficulty in the efficiency measurement, this paper suggests a way to measure the operational efficiency of EVCSs based on data envelopment analysis (DEA). The DEA model is formulated as constant returns of output-oriented model with five types of inputs, four of them are the numbers of floating population and nearby charging stations, distance of nearby charging stations and traffic volume as desirable inputs and the other is the traffic speed in congestion as undesirable one. Meanwhile, the output is given by the charging frequency of EVCSs in a day. Using real-world data obtained from reliable sources, we suggest operational efficiencies of EVCSs in Seoul and discuss implications on the development of electric vehicle charging network. The result of efficiency measurement shows that most of EVCSs in Seoul are inefficient, while some districts (Nowon-gu, Dongdaemun-gu, Dongjak-gu, Songpa-gu, Guro-gu) have relatively more efficient EVCSs than the others.

해사법원이 없는 우리나라에서는 일반법원이 해사사건을 해결하고 있지만, 중요한 해사사건에 대한 재판은 영국의 법원에서 진행되기 때문에 막대한 자본이 유출됨으로써 국가적으로 관련 산업의 경쟁력이 약화되고 있는 실정이다. 이와 같은 상황에서 법조계와 학계는 물론 관련 업계도 해사법원설립의 필요성에 대해서는 공감대를 형성하여 활발한 활동을 펼치고 있지만, 해사법원의 관할문제나 설립 장소에 대해서는 의견대립이 있다. 그리고 해사법원에 형사관할권을 포함하는 문제도 논란의 대상이 되고 있는데, 형사관할권은 형사재판의 편의성뿐만 아니라 피고인의 방어권 보장과도 관련되어 있기 때문이다. 이러한 논의에도 불구하고 해사법원의 형사관할권을 인정해야 하는 이유는 우선 일반법원에서 해사형사사건을 재판하고 있는 현행제도가 전문성의 부족으로 국민의 신뢰를 충분히 받지 못하고 있기 때문이다. 그리고 피고인의 방어권 보장과 관련된 문제는 피고인이 제1심 재판의 관할법원을 일반법원과 해사법원 가운데서 선택할 수 있도록 함으로써 해결이 가능하기 때문이다. 더 나아가 중국에서는 이미 해사법원이 해사형사사건도 관할하고 있다는 사실을 고려할 때 우리나라에서도 해사법원의 형사관할권을 인정할 여지가 충분히 있는 것으로 보인다. 이와 같이 해사형사사건도 해사법원을 통해 해결함으로써 재판의 전문성뿐만 아니라 관련 산업의 경쟁력도 제고할 수 있을 것으로 보인다.

Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during In Vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

For air express service providers offering various express delivery services such as overnight delivery and next-business day delivery services, establishing quickly cargo loading plans is one of important issues owing to the characteristics of air express business, i.e., a short amount of time is available to complete all cargo loading operations before flight departure after receiving air express containers, pallets and bulks. On the other hand, one of major concerns in the air cargo loading planning is to make a plan that insures the stability of an aircraft to avoid take-off, flight, and landing accidents. To this end, this paper considers an air cargo loading planning problem, which is the problem of determining locations in the aircraft cargo space where air containers, pallets and bulks to be loaded while insuring the aircraft stability, motivated from DHL and Air Hong Kong. The objective of the problem is to maximize the total revenue gained from loading air express containers, pallets and bulks. To solve the problem, this paper suggests a simulated annealing algorithm to overcome impracticality of the integer programming model developed by a previous study requiring excessive computation time. The results of computational experiments show that the heuristic algorithm is a viable tool for establishing express cargo loading plans as giving robust and good solutions in a short amount of computation time. Scenario analyses are performed to investigate the effect of the current activities of air express carriers on the revenue change and to draw practical implications for air express service providers.

Fruit peels are potential sources of proteases which can help to the proteins digestion and the encapsulation technique is widely used in food industry, which preserves active ingredients in food products by forming coating membrane. In the present study, the effect of addition of encapsulated fruit peels (kiwi, T1; pineapple, T2; pear, T3; fig, T4) on the quality stability of press ham was studied during refrigerated storage (1, 15 and 30 days). In the first experiment, the press ham was prepared with either encapsulated fruit peels or fruit peels powder (without encapsulation) and we observed that the press ham formulated with fruit peels powder showed a texture defect, but no change with encapsulated fruit peels probably due to the proteolytic activity of proteases were prevented by the encapsulation. In the second experiment, the press ham were made with 0.1% encapsulated fruit peels or normal press ham (control) and stored at different days as mentioned above. Our results revealed that T2 showed the highest moisture content, while the control had the highest fat content. The press hams made with all types of encapsulated fruit peels had significantly higher hardness value than the control throughout the storage. Additionally, higher taste, texture and acceptability scores were found in the press ham with encapsulated fruit peels than those in the control. Overall, it is suggested that the addition of encapsulated fruit peels into meat products may enhance the protein digestion and absorption during the digestive processes without the negative effect on texture and sensory traits.

In order to investigate the effects of antioxidants on the culture of mouse preantral follicles in vitro, we examined the effects of taurine, glutathione and catalase on their growth and maturation. Addition of taurine was not effective on the survival of preantral follicles. However, metaphase II rates of oocytes within preantral follicles were signifi-cantly higher in 1 mM treated group than in control and 10 mM treated group (p<0.05). Glutathione did not improved the rates of survival and metaphase II oocytes. However, metaphase II rates of oocytes progressively decreased with increasing glutathione concentration. Catalase also showed that the rates of survival and metaphase II oocytes pro-gressively decreased with increasing concentration. Especially, all of preantral follicles cultured in medium containing 100 IU/ml catalase were degenerated. These results suggest that low concentraion of taurine, as an antioxidant, have positive effect on the culture of mouse preantral follicles in vitro.

The aim of the present study was to evaluate the effect of ultimate pH of semimembranosus muscle on quality characteristics of dry-cured ham. The sample selection was determined based on ultimate pH of semimembranosus muscle, and samples were then classified into three groups: A (pH 5.61±0.09), B (pH 5.86±0.06) and C (pH 6.13±0.09). Our results depicted that the ultimate pH had a significant effect on the quality characteristics of dry-cured ham. Particularly, as the ultimate pH increased, the pH values of the dry-cured hams significantly (p<0.05) increased while weight loss decreased. Significantly (p<0.05) higher CIE a*, b* and chroma values were observed in the dry-cure hams from the samples with lower ultimate pH. Additionally, the values of some texture characteristics (e.g., hardness and chewiness) significantly (p<0.05) decreased as the ultimate pH increased. However, no significant differences among the three pH groups were observed for water activity, salinity, volatile basic nitrogen (VBN), calorie as well as sensory properties of dry-cure hams (p>0.05). These results clearly demonstrate that the ultimate pH of semimembranosus muscle is more related to quality characteristics than sensory attributes of dry-cured ham.

It is generally accepted that chronic stress impairs female reproduction. It negatively affects ovarian function and the number of ovulated oocytes. Chronic stress lowers the number of retrieved oocytes. Ovarian follicular development is regulated by both pituitary-derived gonadotropins and intraovarian regulatory factors. The main corticosteroids are cortisol, cortisone, 11-deoxycortisol and corticosterone, cortisol being one of the most commonly used welfare and stress physiological indicator. In this study, we investigated the effect of cortisol level on progesterone patterns and ovulation in the dog. Cortisol and progesterone level of serum were analyzed by radioimmunoassay. The day of ovulation was considered as the day when serum progesterone concentration was 6.0∼8.0 ng/ml. In vivo dog oocytes were collected by flushing oviducts of mixed-breed bitches at three days after ovulation. We classified dogs as having group 1 (cortisol level, 0 ≤ or < 2 μg/dl), group 2 (corisol level, 2 ≤ or < 4 μg/dl), group 3 (cortisol level, 4 ≤ or < 6 μg/dl) and group 4 (cortisol level, 6 μg/dL ≤). The patterns of progesterone were not different in four cortisol groups. The average numbers of retrieved oocytes was not different in four cortisol groups. These results suggest that different cortisol levels on estrus dogs do not affect ovulation, number of ovulated oocytes and progesterone changes.

The objective of this study was to monitor health conditions of four genetically identical somatic cells cloned Labrador retriever puppies by estimation of body weight and analysis of hematologic and serologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and four live puppies were delivered by Caesarean section. The cloned Labrador retrievers were genetically identical to the nuclear donor dog. The body weight of clone-1, -2, -3, and -4 was increased from 0.66, 0.40, 0.39, and 0.37 kg at birth to 6.2, 6.6, 6.2, and 6.0 kg at 8 weeks of age, respectively. Although clone-4 had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned puppies were not different when compared to reference range. In serological analysis, Glucose, ALP and inorganic phosphate level of four cloned puppies was significantly higher than the reference ranges. However, there was no significant difference among four cloned dogs. This study suggests that cloned puppies derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological and serological parameters.

The purpose of this study was to assess follicular viability and competence through in vitro culture of preantral follicles isolated from vitrified mouse whole ovaries. Mouse preantral follicles were enzymatically isolated from vitrified-warmed and fresh ovaries and cultured for 10 days followed by in vitro oocyte maturation. In vitro matured oocytes were fertilized and cultured to the blastocyst stage. Five minutes pre-exposure to vitrification solution of whole ovaries had significantly higher (p<0.05) oocyte survival and maturation rates than between 10 min exposure groups. Oocyte diameter was significantly smaller (p<0.05) in the 5 and 10 min exposure groups (69.4±2.8 and 67.8±3.1) when compared to that of control group (71.7±2.1). There was no statistical significant difference in blastocyst development rates between vitrification group (8.6%) and the fresh control group (12.0%). The mean number of cells per blastocyst was significantly lower (p<0.05) in the vitrification group (41.9±20.2) than in the fresh control group (55.1±22.5). The results show that mouse oocytes within preantral follicles isolated from the vitrified whole ovaries can achieve full maturation, normal fertilization and embryo development.

Limited success of somatic cell nuclear transfer(SCNT) is attributed to incomplete reprogramming of transferred donor cell. Several approachs, such as histone deacetylase inhibitors and DNA methyltransferase inhibitors have been used to improve the efficiency of somatic cell nuclear transfer. Recently, it is reported that pre-treatment of somatic cells with undifferentiated cell extract, such as embryonic stem cell and mammalian oocytes is an attractive alternative ways to reprogramming control. The aim of this study was to evaluate the early development of porcine cloned embryos produced with porcine ear skin fibroblasts pre-treated with extract from porcine induced pluripotent stem cell (iPSC). For transport of porcine iPSC extract into cultured porcine ear skin fibroblasts, the ChariotTM reagent system was used. Treated cells were cultured for 3 days, and used for the analysis of histone H3K9 acetylation and SCNT The acetylation status of H3K9 was increased in cells treated with iPSC extract and cultured for 3 days compared with control. But, no significant difference was observed between the extract treated and control groups. After SCNT. no difference was observed in the rate of fusion (86.6% vs 86.2%) and embryo cleavage (86.6% vs 87.1%) between the extract treated and control groups. Also, no significant difference was noted in blastocyst rates (23.4% vs 28.4%) as well as cell numbers (43.8±10.8 vs 41.2±11.6) with extract treated group compared with control group. Overall apoptosis rate in blastocyst was not differences between the extract treated and control groups (4.6±3.5% vs 6.0± 5.8%). However, blastocyst rate with high apoptotic cells(>10% appototic cells) was significantly lower in extract treated group when compared with control group (7.1% vs 21.8%).. Our results demonstrated that pre-treatment of porcine ear skin fibroblasts using porcine iPSc extract had beneficial effect on the decreasing apoptosis in the blastocyst cultured in vitro, although there was no effect on the embryonic development.

Despite of the absence of hyperacute rejection and acute humoral xenograft rejection, the organ graft of the a1,3-galactosyltransferase (GalT) gene knockouted (KO) and complement regulatory protein (CRP) expressing pig into a nonhuman primate is rejected by development of a thrombotic microangiopathy and/or a consumptive coagulopathy. Thus further introduction of genes to overcome the coagulation incompatibilities between pig and primate under GalT KO/CRP genetic background has been strongly suggested. CD73 (ecto-5'-nucelotidase) is an enzyme attached via a glycosyl phosphoinositol anchor to the extracellular membrane of endothelial cells, which catalyses the hydrolysis of adenosine triphosphate to adenosine. Loss of activity of CD73 results in activation and aggregation of platelets by a reduced capacity to convert nucleotides to adenosine. In previous study, we reported generation of GalT KO fibroblasts concurrently expressing membrane cofactor protein and produced cloned pigs by nuclear transfer of the fibroblast cells (1). In this study, we constructed a vector for expression of human CD73 under control of promoter of pig Icam2 gene expressed specifically at endothelial cells. This vector was introduced into porcine fibroblasts using the nucleofection technology, by which we had forty three fibroblasts clones carrying pIcam2- CD73 vector. Somatic cell nuclear transfer resulted in generation of two transgenic piglets survived.

Immunological rejection of the organ grafted onto a primate arises from two antibody mediated processes, hyperacute rejection (HAR) and acute humoral rejection (AHR). Functional ablation of α1,3-galactosyltransferase (GalT) and concurrently overexpression of complement regulatory proteins are known to inhibit HAR and AHR. In previous study, we reported that production of porcine male fibroblasts harboring a MCP expression cassette targeted to GalT locus. In this study, we constructed a different MCP expression cassette, in which the EF1α promoter regulates MCP expression and internal ribosome entry site-mediated neomycin resistance gene expression. Subsequently, this cassette was inserted between the left and the right homologous arms to target exon 9 of the GalT gene. Female fibroblasts were isolated from ear skin of 10 days old miniature pig, and used for nucelofection of the the construct for MCP expression at GalT locus. PCR analysis showed that four clones of forty neomycin resistant clones carry MCP expression cassette at exon 9 of the GalT gene. Two clones analyzed downregulated GalT expression, as determined by quantitative reverse transcriptase polymerase chain reaction. Flow cytometry analysis showed that MCP was efficiently expressed at the cell surface.

Culture of preantral follicles has important biotechnological implications through its potential to produce large quantities of oocytes for embryo production and transfer. The objective of this study was to determine the comparison of different isolation method of mouse preantral follicles, and to examine in vitro development of mouse preantral follicles isolated by different method. Preantral follicles were mechanically or enzymatically extracted from mouse ovaries. Mechanical isolation method used fine gauge needles and enzymatic method of isolating follicles used 1 mg/ml collagenage (Type IA) and 0.2 mg/ml DNase Ⅰ in Leibovitz L-15 medium. The solution containing Leibovitz L-15 medium, enzyme and ovary fragments was incubated at 37℃ for 30 min. The selection criteria are as follows: primary follicle of 75 to 99 μm, early secondary follicle of 100 to 125 μm and late secondary follicle of 126 to 150 μm in diameter. The recovered preantral follicles were cultured for 10 days in alpha-minimal essential medium (α-MEM) + 5% FBS + ITS + 100 mIU/ ml FSH. The collected primary follicles by enzymatic treatment were higher than mechanical method. Others stage preantral follicle by mechanical isolation were higher than enzymatic method. After 10 days of culture, no statistical differences were shown in survival rates of preantral follicle among the 2 culture groups. The metaphase Ⅱ rates of the oocytes were significantly higher (p<0.05) in mechanical method (17.8%) than in enzymatic method (5.1%). These results suggest that the isolation method of choice depends on the target stage preantral follicles and mechanical isolation is an optimal method of preantral follicles in a culture of mouse preantral follicle.

Although the National Institute of Health (NIH, USA) miniature pigs were developed specifically for xenotransplantation, the cloning efficiency is still very low. To increase the efficiency, an advanced somatic cell nuclear transfer (SCNT) method may need. In the present study, we report the productions of genetically modified cloned pigs using the frozen-thawed donor cells without culture before SCNT. Fibroblasts were isolated from an ear skin of a 10-day-old NIH miniature pig. The fibroblast cells were genetically modified with the human CD73 (hCD73). For SCNT, somatic cells transfected with hCD73 were used as donor cells. The survival rate of the somatic cells was significantly higher in 0 h (95%) compared with 1 h (81%) after thawing (p<0.05). We obtained the pregnancy (38.9%, 7/18) and delivery (11.1%, 2/18) rate, respectively. Totally 7 genetically modified cloned piglets were delivered. Among them, 2 piglets were survived and 5 piglets were born stillbirth. The healthy 2 piglets are still survived (≥6 months).

This study investigated whether the addition of porcine sperm cytosolic factor (SCF) at fusion/activation affects in vitro development of porcine parthenogenetic(PA) and nuclear transfer (NT) embryos. To determine the optimum concentration of SCF, control group of oocytes was activated with 0.3M mannitol (1.0 mM CaCl2 ․ 2H2O), other three groups of oocytes were parthenogentically activated with the fusion medium (0.1mM CaCl2 ․ 2H2O) supplemented with 100, 200 or 300 μg/ml SCF, respectively. Matured oocytes were activated with two electric pulses (DC) of 1.2 kv/cm for 30 μsec. The activated embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Oocytes activated in the presence of SCF showed a significantly higher blastocyst rate than control (p<0.05). Apoptosis rate was significantly lower in 100 μg/ml SCF group than other groups (p<0.05). Cdc2 kinase activity in control and SCF treatment group of oocytes was determined using MESACUP cdc2 kinase assay kit at 1, 5, 10, 15, 30, 45 and 60 min after activation. Cdc2 kinase activity was significantly decreased (p<0.05) in SCF group than MII oocytes or control within 5 min. For NT embryo production, reconstructed oocytes were fused in the fusion medium supplemented with 0.1 mM CaCl2 ․ 2H2O (T1), 1.0 mM CaCl2 ․ 2H2O (T2) and 0.1 mM CaCl2 ․ 2H2O with 100 μg/ml SCF (T3). Fused embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Developmental rate to blastocyst stage was significantly higher in T3 than other groups (23.0% vs. 13.5 to 15.2%) (p<0.05). Apoptosis rate was significantly lower in T3 than T1 or T2 (p<0.05). The relative abundance of Bax-α/Bcl-xl was significantly lower in in vivo or SCF group than that of control (p<0.05). Moreover, the expression of p53 and caspase3 mRNA was significantly lower in in vivo or SCF group than that of control (p<0.05). These results indicate that the addition of SCF at fusion/activation might improve in vitro development of porcine NT embryos through regulating cdc2 kinase level and expression of apoptosis related genes.

This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM SrCl2 in Ca-free CZB medium in the presence of 5 μg/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6～7 week-old mice(53.3%) than in oocytes from 8～9(80.9%) and 10～11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium (51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6～7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.