This paper compared the consistency of the Environment Stress(ES) model and the Potential Risk Assessment Model (PARK model, which was developed based on a Korean mariner risk perception) for the Busan adjacent waterway. Evaluation of accuracy and comparison of these two models have been made by Vessel Traffic Service (VTS) officers in the Busan VTS Centre. The assessment results of Busan waterway show that the PARK model is more consistent than the ES model as follows. (1) The difference between assessment results applying ES model and PARK model with risk degree of VTSOs were 34% and 5% respectively in six typical traffic situations. (2) The assessment using PARK model is more suitable and identical with the VTSOs opinion in his or her duty time.

This study was conducted to evaluate the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To determine the movement of degradable HGC nanoparticles in the body, 20 mg/kg of lutetium177-labeled HGC (Lu177-HGC) with the size ranging from 320 to 400 nm was injected intravenously into ICR mice, and the amount of radioactivity remaining in blood and several organs was measured at various time points during the period of 5 days. In the pharmacokinetics analysis using the Lu177 radioisotope, the free Lu177 was mainly distributed and accumulated in the order of kidney>liver>lung at 1 day after the injection of the radioisotope. However, the Lu177-HGC showed a high distribution of nanoparticles in the order of liver>spleen>kidney during the experimental period of 5 days. These results would provide a basic pharmacokinetics for the use of HGC as a drug carrier in drug delivery system.

The ultimate goal of this study is to assess the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To understand the movement of degradable nanoparticle HGC in the body, we intravenously injected a dose of 20 mg/kg of Cy5.5-labeled HGC with size ranging from 320 to 400 nm into ICR mice, and measured the amount of fluorescence remaining in blood and several organs at various time intervals. In blood, the level of Cy5.5-labeled HGC was the highest at 15 min, then after 30 min it decreased rapidly and reached a plateau form 30 min to 28 days. In the tissue we confirmed the presence of nanoparticles at high levels in the order of kidney>liver>submandibular gland until 28 days after injection. However, we did not find the presence of the particles in the brain or testes. These results will provide basic information on HGC as a drug delivery agent.

Mimosa pudica (미모사), Argemone mexicana (멕시코 가시양귀비) Leucus aspara (꿀풀과 일종), Polygonum hydropiper (여뀌), Blumea lacera (국화과 일종) 등 5종의 식물 헥산추출물들의 팥바구미 성충에 대한 살충, 성충우화억제력 및 녹두 종실피해 방제력이 검증되었다. 그 결과, 밭바구미 성충 살충력은 35-69%이었으며, 녹두에 추출물을 처리한 후 성충우화 방제율은 33-63%, 종실 피해 방제율은 13-49%이었다. 추출물의 농도가 증가할수록 살충력, 성충우화 및 종실피해 방제율 등이 증가하는 것으로 나타났다. 이와 같이 5종의 식물 헥산추출물들은 밭바구미 친환경 방제제로 이용가능성이 있을 것으로 판단된다.

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

Antibiotic Detection Kit (Combination I), a lateral flow immunoassay (LFIA) developed for the detection of antibiotic residues in milk, was utilized for the analysis of antibiotic residues in the muscle tissue of olive flounder. After 60-min treatment by dipping in water dosed with ampicillin (200-g/ton water), the residue depletion of ampicillin was investigated in 25 cultured olive flounder (Paralichthys olivaceus). Muscles of fish were sampled on the 1st, 2nd, 3rd, 4th and 5th day after drug treatment. The concentration of ampicillin in the muscle was determined by LFIA. The absorbance ratio of the sample to the control blank (Bs/Bo) was employed as an index to determine the muscle residues in olive flounder. To investigate the recovery rate, standard solutions were added to muscle samples to give final concentrations in the muscle of 4 and 8 ng/ml. The recovery rates of all spiked samples were > 96% of the spiked value. Ampicillin was detected in the muscle of fish treated with the drug until the 2nd day of the withdrawal period. The present study showed that the LFIA can be easily adopted to predict ampicillin residues in tissue of farmed fishes.

During January-November 2012, a total of 2,041 quarter milk samples were collected from dairy cattle of 82 dairy farms nationwide. About 42% (870/2,041) of the samples that had somatic cell counts (SCC) of ≥ 200,000 cells/ml were subjected to microbiological examination. No bacteria was isolated from 95 of 870 (10.9%) samples. Among 1,237 bacteria isolated from the rest 775 samples, 1,085 were identified with VITEK: more than half (52.1%, 645/1,237) of the isolates were gram negative bacillus. Gram positive cocci including Staphylococcus accounted for 35% of the isolates and almost none of gram positive bacilli isolated. Excluding Coagulase-negative Staphylococcus (CNS), the most frequently isolated bacterial species was Escherichia coli (11.2%, 138/1, 237), followed by Klebsiella pneumoniae (8.1%,100/1,237), Staphylococcus aureus (7.1%, 88/1, 237), Enterobacter cloacae (6.0%, 74/1, 237), and Serratia marcescens (3.5%, 43/1, 237). The most common resistance of S. aureus was to penicillin (77.4%) and ampicillin (73.0%), while no resistance was observed against gentamicin and cephalothin. Although CNS presented resistance to all antimicrobials tested but the most prevalent resistance was to penicillin (35.6%) and ampicillin (37.0%). The pattern of antimicrobial resistance observed in CNS was similar to that of S. aureus, but the rates were much lower than those of S. aureus. E. coli also showed resistance to all the antimicrobials tested, although the rates were not very high. The highest resistance of E. coli was to cephalothin (39.4%) and ampicillin (36.2%), while most of the strains (98.0%) showed sensitivity to amikacin. The results of this study provide information on current situation of bovine mastitis in Korea.

Mealworms, Tenebrio molitor (L.) is used as an important animal feed additive for growth promotion and health management, but potentially exposes to fungal infection. In this work, virulence of two species of entomopathogenic fungi against the insect, and the relationship between abiotic features and virulence were investigated. Secondly our consideration was also given to the effect of chemical fungicides on conidial germination for risk control. Between Beauveria bassiana (Bb) and Metarhizium roberstii (Mr) (previously M. anisopliae), Bb isolates had much higher virulence (~100% mortality in 3~4 days after the treatment), rather than Mr isolates in laboratory assays. Next, fungus-treated mealworms were kept at wheat bran at 20, 25, 30 and 35℃ with 3, 6, 9 times of water spray to the feeds for set-up of different humidity conditions. Inoculation of fungi to mealworms was conducted by fungal spray and feeding methods, which resulted in higher virulence in feeding method. In the feeding method, all temperature treatments except 35℃ showed high virulence against mealworms, but any significant relationship between virulence and humidity was not observed. In the chemical fungicide screening, fluazinam (CAS No. 79622-59-6) and mancozeb (8018-01-7) significantly inhibited the germination of Bb and Mr conidia. This work suggest that contamination of wheat bran with fungal pathogens, particularly B. bassiana may induce mycosis of mealworms, but introduction of effective fungicides possibly reduce fungal infection.

An assessment was made of the toxicity of hiba, Thujopsis dolabrata var. hondai, oil and 24 oil constituents and control efficacy of four experimental spray formulations containing the oil (0.5. 1, 2, and 3% sprays) to adult American house dust mite (AHDM), Dermatophagoides farinae, and copra mite (CM), Tyrophagus putrescentiae. Results were compared with those of three conventional acaricides benzyl benzoate, N,N-diethyl-3-methylbenzamide (deet), and permethrin. Based on 24 h LC50 values, (–)-thujopsene was the most toxic constituent against AHDM (9.82 μg/cm2) and CM (10.92 μg/cm2) and the toxicity of the compound was nearly identical to that of benzyl benzoate (9.33 and 10.14 μg/cm2). High toxicity was also observed with (–)-(E)-pinocarveol, carvacrol, (+)-terpinen-4-ol, β-thujaplicin, cedrol, α-terpineol, (+)-ledene, thymol, and 2,4-di-tert-butylphenol against both mite species (LC50, 11.92–19.19 and 12.49–22.97 μg/cm2). These constituents were more toxic than deet (LC50, 35.53 and 38.42 μg/cm2). Hiba applied as 2 and 3% sprays provided >95% mortality against both mite species, whereas permethrin (cis:trans, 25:75) 2.5 g/L spray treatment resulted in ca 11% mortality. Global efforts to reduce the level of highly toxic synthetic acaricides in indoor environments justify further studies on hiba oil preparations containing the constituents described as potential contact-action fumigants or lead molecules for the control of mite populations.

The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The dissemination process of agricultural research and development (R&D) results has somewhat different characteristics from that of typical R&D results. However, these characteristics are not adequately considered on the basis of an examination of the current performance system, the resulting management plans, and strategies for the application and dissemination of the results of agricultural R&D in Korea. The performance evaluation indicator exposed the problem of the inadequate consideration of the characteristics of each of these areas, particularly the lack of unified R&D-related institutions and the inadequacy of the system to monitor outcomes. To address these shortcomings in the agricultural R&D programs in Korea, the policies pertaining to agricultural R&D performance, results management, and dissemination in the U.S. and Japan were examined. Based on these investigations, we proposed strategies to improve the agricultural R&D policies in Korea.

This study was to examine on the respiratory variables, heart rate and muscle activity between the static recovery and dynamic recovery after progressive resistance exercise to maximal point. Subjects were 15 students enrolled in N University. All were tested two times (static recovery and dynamic recovery) and were requested to perform a walking on a treadmill after progressive resistance exercise to maximal point. Electromyography(EMG) was used to monitor the muscle activity(TA: Tibialis Anterior, MG: Medial Gastrocnemius) during gait. CPEX-1 was used to measure the respiratory variables and heart rate. The dynamic recovery group was shown the significant lower heart rate than that of static recovery group at during gait. Respiratory rate showed statistically a significant difference. Electromyography(RMS, root mean square) showed a non-significant difference. But the dynamic recovery group of muscle activity was found highly in TA and MG. This study indicated that the dynamic recovery method evidenced more faster than the static recovery method. And this type of dynamic rest by walking can be a help of recovery after exercise.

Oleanolic acid is a natural triterpenoid that exists widely in foods and some medicinal herbs. The purpose of this study was to determine the antimicrobial activity of oleanolic acid against Streptococcus mutans strains isolated from a Korean population. Antimicrobial activity against these bacteria was evaluated by minimal inhibitory concentration (MIC) and time kill curves. The tolerance of human gingival fibroblasts and human periodontal ligaments to oleanolic acid was tested using a methyl thiazolyl tetrazolium (MTT) assay. The MIC90 value of oleanolic acid for both S. mutans and S. sobrinus isolated from Koreans was 8μg/mℓ. Oleanolic acid showed bactericidal effects against S. mutans ATCC 25175T and S. sobrinus ATCC 33478T at 1 × MIC(8μg/mℓ) and had no cytotoxic effects against KB cells at this dose. The results suggest that oleanolic acid could be useful in the future development of oral hygiene products for the prevention of dental caries.

This study was to investigate the effects of upper and lower limb composing patterns of PNF(proprioceptive neuromuscular facilitation) on the static balance ability by 20 subjects for 6 weeks. This study was measured left one leg standing and right one leg standing with closed eyes on Good Balance system. These results led us to the conclusion that the mean speed of X, Y direction, COP(center of Pressure) velocity moment showed a statistical decrease when applying post-exercise. The above results from this study indicated that upper and lower limb composing patterns of PNF exercise has improved the static balance ability. As a result, this study showed that upper and lower limb composing patterns exercise improve the ability of balance in young adults. Based on this study, it may be applied to old people.

Purpose: The purpose of this study was to estimate the effects of subliminal music with balance imagery training on balance and concentration. Methods: The participants were 45 seniors in an undergraduate school in Korea. The subliminal music with balance imagery training intervention was carried out for 20 minutes. Other interventions were also carried out for 20 minutes. 12 seniors(Group A) listened to subliminal music with balance imagery training, 12 seniors(Group B) listened to subliminal music, 11 seniors(Group C) received balance imagery training, and 10 seniors(Group D) had no intervention(Control group). The grid test is related to measured levels of concentration intensity. Romberg one legged standing test was carried out for 30 seconds. The collected data was analyzed by one-paired t test and one way ANOVA using the SPSS Windows 12 ver. program. Results: The major findings of this study were as follows: Concentration levels of Group A and C improved, and balance levels of Group C and D improved. There was a statistically significant decrease in concentration between Group A and B, Group A and C after intervention. Conclusion: These findings suggest that listening to subliminal music with balance imagery training may be useful in managing concentration in seniors. So it provides basic information for further concentration on improving education on music with balance imagery training.