Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cul-tured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn’t stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.

Mushrooms are considered not only as food but also for source of physiologically beneficial medicines. The culinarymedicinal mushrooms may important role in the prevention of age-associated neurological dysfunctions, including Alzheimer’s and Parkinson’s diseases. Hericium erinaceus (H. erinaceus), is edible mushrooms, is a parasitic fungus that grows hanging off of logs and trees and well established candidate for brain and nerve health. H. erinaceus contains high amounts of antioxidants, beta-glucan, polysaccharides and a potent catalyst for brain tissue regeneration and helps to improve memory and cognitive functions. Its fruiting bodies and the fungal mycelia exhibit various pharmacological activities, including the enhancement of the immune system, antitumor, hypoglycemic and anti-aging properties. H. erinaceus stimulates the synthesis of Nerve Growth Factor (NGF) which is the primary protein nutrient responsible for enhancing and repairing neurological disorders. Especially hericenones and erinacines isolated from its fruitin body stimulate NGF, synthesis. This fungus is also utilized to regulate blood levels of glucose, triglycerides and cholesterol. H. erinaceus can be considered as useful therapeutic agents in the management and/or treatment of neurodegeneration diseases. However, this review focuses on in vitro, in vivo and clinical trials for neurodegerative disease.

This study was conducted to investigate the community structure and species distribution of ground beetle assemblages in western part of Yeongwol-gun, Korea. Ground beetles were collected by pitfall traps from 9 study sites during July to October in 2013. A total of 30 species were identified from 4,439 collected ground beetles. Two dominant species, Synuchus nitidus (1,916 individuals) and Synuchus cycloderus (1,900 individuals) were occupied 85.8% of total. Interestingly, Pterostichus orientalis orientalis and Synuchus melantho were only collected in relative high altitude above 350 m, such as Beopheung-ri, Unhak-ri, and Yongseok-ri. Large-sized beetles, such as Damaster, Leptocarabus, and Carabus species, were generally more collected from Suju-myeon (Beopheung-ri and Unhak-ri) and Jucheon-myeon (Docheon-ri and Yongseok-ri) as well-conserved area, but not occurred in Yeongwol-eup and Hanbando-myeon, which forests may have lower habitat quality for these species. These differences also lead to increasing of dissimilarity among study sites, which were diverged into 3 groups at 53.0% of similarity. In particular, species composition of Samok-ri was significantly different at 8.2% of similarity because of difference in habitat characteristics as river side. This study has an important value to provide the fundamental information of inventory and monitoring in Yeongwol-gun. For detail mapping of species distribution in Yeongwol-gun, however, further studies are necessary, especially eastern parts of Yeongwol-gun.

Epigeic spiders (Araneae) were surveyed in apple orchards under different farming practices (organic and conventional) in Muju-gun, Jeollabuk-do, Korea. The study was conducted from April to October in 2013 with 2 weeks interval using pitfall traps. Collected spiders were identified as 20 species of 6 families from 819 individuals in organic orchardⅠ, 19 species of 10 families from 759 individuals in orchardⅡ and 16 species of 8 families from 209 individuals in conventional orchard for from 1,787 captured spiders. Biodiversity was analyzed with species richness, abundance and Shannon diversity index (H΄). Species richness was statistically higher in organic orchard than in conventional orchard. Organic orchardⅡ comprised the greatest abundance followed by organic orchard and conventional. But species diversity was the highest in organic orchardⅡ, intermediate in conventional orchard and the lowest in orchardⅠ. Lycosidae was the most dominant family in all orchards regardless farming practices. However, Linyphiidae dominated in organic orchards only. The most dominant species were Erigone koshiensis (Linyphiidae) in organicⅠ, Pardosa laura (Lycosidae) in organicⅡ and Arctosa pungcheunensis (Lycosidae) in conventional orchard.

This peptide has antibacterial activity against several Gram-positive and Gram-negative bacteria. BmCecB1 is antimicrobial peptides from Bombyx mori and belongs to cecropin family. Antimicrobial peptides are important components of the innate immune systems in all living organism. To produce the BmCecB1 antimicrobial peptide, we constructed transgenic silkworm that expressed BmCecB1 gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 600 eggs of bivoltin silkworms, Baegokjam. In total, 49 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 22 broods, and we selected 2 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 11%. We identified 9 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a BmCecB1 as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that BmCecB1 possesses high antibacterial activities against Gram-negative bacteria.

Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.

Screening for antimicrobial peptide genes in the immune-induced Antheraea yamamai larvae led to the identification of a novel antifungal moricin-like peptide (MLP10) gene. The complete MLP10 cDNA is comprised of 403 bp with 174 bp open reading frame encoding a 58 amino acid precursor that contains a putative 23-residue signal peptide, a 2-residue propeptide and a 33-residue mature peptide. The deduced amino acid sequence of MLP10 has 26∼52% identity to those of moricin-related peptides from lepidopteran insects. The MLP10 was highly expressed in E. coli BL21(DE3) by fusing with ketosteroid isomerase (KSI) to avoid the cell death during induction. The resulting expressed KSI-MLP10 fusion protein was in a insoluble form. Recombinant MLP10 was released by cleavage of the fusion protein with cyanogen bromide (CNBr). Subsequently, we purified pure active MLP10 by FPLC chromatography, and 5.2mg of MLP10 was obtained from 1L culture medium. The purified MLP10 was prevented the growth of candida albicans at 6.25 uM, and was also active against gram negative and gram positive bacteria. This potent antimicrobial activity suggests that MLP10 may play a role in the immune response of A. yamamai.

Selecting surrogated species is the first step for environmental risk assessment of GM crop on non-target arthropods. To select surrogated species considering the domestic environment, we surveyed the arthropod fauna in five main localities (Changnyeong-gun in Gyeongsangnam-do, Goesan-gun in Chungcheongbuk-do, Gokseong-gun in Jeollanam-do, Hongcheon-gun in Gangwon-do and Hwaseong-si in Gyeonggi-do) in which maize was cultivated with large scale. Maize is one of the main imported GM crop in Korea. Arthropods were surveyed with yellow sticky traps and pitfall traps from May to August with a week interval considering the crop growing season in 2012 and 2013. A total of 135 species of 51 families in 13 orders were identified from the 17,470 arthropods. Six orders (Coleoptera, Araneae, Hemiptera, Diptera, Hymenoptera and Lepidoptera) were dominant in species richness and three orders (Collembola, Thysanoptera and Diptera) in abundance, representing 84% and 86% of the total, respectively. In the composition of fuctional guilds, herbivore group was the most dominant followed by predator group occupying 40% and 37% of the total abundance.

Pure MgH2 was milled under a hydrogen atmosphere (reactive mechanical grinding, RMG). The hydrogen storage properties of the prepared samples were studied at a relatively low temperature of 423 K and were compared with those of pure Mg. The hydriding rate of the Mg was extremely low (0.0008 wt% H/min at n = 4), and the MgH2 after RMG had higher hydriding rates than that of Mg at 423 K under 12 bar H2. The initial hydriding rate of MgH2 after RMG at 423 K under 12 bar H2 was the highest (0.08 wt% H/min) at n = 2. At n = 2, the MgH2 after RMG absorbed 0.39 wt% H for 5 min, and 1.21 wt% H for 60 min at 423K under 12 bar H2. At 573 K under 12 bar H2, the MgH2 after RMG absorbed 4.86 wt% H for 5 min, and 5.52 wt% H for 60 min at n = 2. At 573 K and 423 K under 1.0 bar H2, the MgH2 after RMG and the Mg did not release hydrogen. The decrease in particle size and creation of defects by reactive mechanical grinding are believed to have led to the increase in the hydriding rate of the MgH2 after RMG at a relatively low temperature of 423 K.

Immune-inducible antimicrobial peptides were produced using transgenic silkworms that expressed Rel family transcription factor, truncated BmRelish1 (BmRelish1t) genes under the control of the BmA3 promoter using the piggyBac vector. BmRelish1t gene contains all domains of Bmrelish: a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acids (AHAA) rich region except the Ankyrin repeat domain (ANK) and the death domain (DD). (1:1) Mixtures of the donor vector (pG-3xP3EGFP-BmA3BmRelish1t) and helper vector were micro-injected into 1,800 eggs of bivoltin silkworms, Baegokjam and EGFP-induced fluorescence was observed for 25 broods of transgenic silkworms under a florescence stereomicroscope. Analysis by real-time PCR indicated that transgenic silkworms expressing BmRelish1t recombinant proteins displayed higher mRNA expression levels of the Bombyx mori antimicrobial peptides such as lebocin, moricin, and nuecin than the normal silkworms. Moreover, transgenic silkworms expressing BmRelish1t showed antibacterial activity against Escherichia coli. We suggest that transgenic expression of BmRelish1t may find useful applications for the production of various antimicrobial peptides at the same time in transgenic silkworms.

Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.

Matrix metalloproteinases (MMPs) have been known to affect to cell migration, proliferation, morphogenesis and apoptosis by degrading the extracellular matrix. In the previous studies, undifferentiated mouse embryonic stem cells (ESCs) were successfully proliferated inside the extracellular matrix (ECM) analog-conjugated three-dimensional (3D) poly ethylene glycol (PEG)-based hydrogel. However, there is no report about MMP secretion in ESCs, which makes it difficult to understand and explain how ESCs enlarge space and proliferate inside 3D PEG-based hydrogel constructed by crosslinkers containing MMP-specific cleavage peptide sequence. Therefore, we investigated what types of MMPs are released from undifferentiated ESCs and how extracellular signals derived from various niche conditions affect MMP expression of ESCs at the transcriptional level. Results showed that undifferentiated ESCs expressed specifically MMP2 and MMP3 mRNAs. Transcriptional up-regulation of MMP2 was caused by the 3D scaffold, and activation of integrin inside the 3D scaffold upregulated MMP2 mRNAs synergistically. Moreover, mouse embryonic fibroblasts (MEFs) on 2D matrix and 3D scaffold induced upregulation of MMP3 mRNAs, and activation of integrins through conjugation of extracellular matrix (ECM) analogs with 3D scaffold upregulated MMP3 mRNAs synergistically. These results suggest that successful proliferation of ESCs inside the 3D PEG-based hydrogel may be caused by increase of MMP2 and MMP3 expression resulting from 3D scaffold itself as well as activation of integrins inside the 3D PEG-based scaffold.

The hydrogen storage properties of pure MgH2 were studied and compared with those of pure Mg. At the first cycle,pure MgH2 absorbed hydrogen very slowly at 573 K under 12 bar H2. The activation of pure MgH2 was completed after threehydriding-dehydriding cycles. At the 4th cycle, the pure MgH2 absorbed 1.55wt% H for 5 min, 2.04wt% H for 10 min, and3.59wt% H for 60 min, showing that the activated MgH2 had a much higher initial hydriding rate and much larger Ha (60min), quantity of hydrogen absorbed for 60 min, than did activated pure Mg. The activated pure Mg, whose activation wascompleted after four hydriding-dehydriding cycles, absorbed 0.80wt% H for 5 min, 1.25wt% H for 10 min, and 2.34wt%H for 60 min. The particle sizes of the MgH2 were much smaller than those of the pure Mg before and after hydriding-dehydriding cycling. The pure Mg had larger hydrogen quantities absorbed at 573K under 12 bar H2 for 60 min, Ha (60 min),than did the pure MgH2 from the number of cycles n=1 to n=3; however, the pure MgH2 had larger Ha (60 min) than didthe pure Mg from n=4 to n=6.

It has been over two decades since the Japanese practice of enforced sexual slavery began to receive widespread attention. Yet despite numerous international efforts to urge Japan to squarely acknowledge its moral and legal responsibility, there has been no meaningful progress to resolve this matter. This work revisits the issue of enforced sexual slavery as it stands today. The Japanese practice of enforced sexual slavery was a clear violation of international law at the time. Therefore, individual victims have valid legal claims for reparation against the Japanese government. The first half of this article reconfirms the illegality of the practice of enforced sexual slavery. The remainder summarizes and vindicates the claims of the victims once again. This research suggests how to remedy the victims’ rights and discusses how to implement reparation. It also contends that Japan owes reparations and legitimate remedial measures to the victims that go beyond monetary compensation in line with the rules of contemporary international law.

The toxicity of red pine needle hydrodistillate, its constituents, and 12 structurally related compounds to third instar larvae of insecticide-susceptible Aedes aegypti and wild Aedes albopictus was examined using a direct-contact bioassay. Results were compared with those of the currently used larvicides fenthion and temephos. The red pine needle hydrodistillate constituents were identified by GC-MS and co-elution of authenticated samples following co-injection. Based on 24 h LC50 values, 3-carene (8.60–9.33 mg/L), limonene (10.34–10.77 mg/L), and thymol (10.65–11.45 mg/L) were the most toxic constituents. The moderate toxicity was produced by (1S)-(–)-α-pinene, p-cymene, geranyl acetate, myrcene, and β -caryophyllene (LC50, 27.62–38.01 and 28.61–39.52 mg/L). Overall, these compounds were less effective than either fenthion (LC50, 0.012–0.022 mg/L) or temephos (LC50, 0.010–0.015 mg/L). In the light of global efforts to reduce the level of highly toxic synthetic insecticides in the agricultural environment, red pine needle hydrodistillate and the compounds described merit further study as potential larvicides for the control of mosquito populations.

In apple orchards under different farming practices (organic and conventional), arthropod community structure was investigated. Arthropods were collected with pitfall traps, band traps, yellow sticky traps and pheromone traps from May to December, 2012 in Danyang-gun, Chungcheongbuk-do, Korea. Biodiversity was analyzed with species richness and Shannon diversity index (H΄). Sampled arthropods were identified 37 families in 14 orders from 21,618 individuals in organic orchard-1 and 33 families in 10 orders from 11,318 individuals in organic orchard-2 and 35 families in 13 orders from 27,711 individuals in conventional orchard. Species richness was relatively highest in organic orchard-1, intermediate in conventional orchard and lowest in organic orchard-2. In ecologically functional group, abundance of pests was higher in conventional orchard than in organic orchards due to the high occurrence of Phyllonorycte ringoniella (Gracillariidae) and Lyonetia prunifoliella (Lyonetiidae). Organic orchards showed higher abundance, species richness and species diversity of epigeic predators and those of parasitoids above ground than conventional orchard. Abundance of detritivores, mainly collembola, were higher in organic orchard than in conventional orchard.

This study was conducted in order to examine the effects of alcohol-free cetylpyridinium chloride drinking water additive and oral gel on clinical parameters related to periodontal disease in beagle dogs. This study was conducted with healthy 15 beagle dogs. Following a professional teeth cleaning procedure, dogs were divided into three groups. Dogs in the control group received nothing, those in the drinking water additive (DWA) group received 800 ml water with 15 ml of alcohol-free cetylpyridinium chloride drinking water additive daily, and those in the Oral gel (OG) group were treated with oral gel containing alcohol-free cetylpyridinium chloride and 0.05% chlorhexidine gluconate daily. Clinical parameters, including plaque index (PI), calculus index (CI), and gingivitis index (GI) were evaluated at two and four weeks. Dogs in the DWA and OG groups had significantly less plaque than dogs in the control group at two and four weeks (P<0.01, P<0.05). And, at four weeks, CI was significantly lower in the OG group compared to the control group (P<0.05). On GI, similar scores were recorded for all groups during the experimental period. No significant difference was observed between the DWA group and the OG group. The effect of alcohol-free cetylpyridinium chloride drinking water additive was similar to the result for alcohol containing cetylpyridinium chloride mouthwash reported in a previous study. The effect in control of periodontal disease was better in the OG group because of additional chlorhexidine gluconate. However, use of drinking water additive will be more convenient for owners; thus, it will be more effective for achievement of long-term results.

In this study, we observed anti-diabetic effects of acid hydrolyzed silk peptides, where the amount of peptides in the total amino acid mixture was strictly regulated. Using in vitro diabetes models, silk peptide-containing amino acid mixtures of 5.60% (G5), 11.30% (G10), 14.50% (G15), and 20.50% (G20) were examined separately in order to determine whether they have biological activities. According to our results, a cytoprotective effect was observed following treatment of interleukin-1β in RINm5f pancreas β-cells. As a consequence, Bax, a pro-apoptotic gene, was down-regulated, while Bcl-2, a pro-survival gene, was retained at normal level. Results of the 4’,6-diamidino-2-phentylindole (DAPI) staining assay confirmed that G20 has a better cytoprotective effect. Insulin release from RINm5f cells showed a significant increase following treatment with G5-G20, suggesting that silk peptide effectively regulated and induced insulin production. Single treatment with G5-G20 resulted in enhanced glucose uptake in L6 skeletal muscle cells. In addition, a higher amount of each group inhibited the activity of α-glucosidase. In summary, these data suggest that silk peptide may have an anti-diabetic effect through protection of pancreas β-cells and enhancement of insulin release, which showed a close association with Type 1 diabetes mellitus (DM), and can improve glucose uptake, which was the major target for therapy of Type 2 diabetes. Taken together, we concluded that acid hydrolyzed silk peptides can be used effectively for control of blood sugar metabolism via improvement of the problematic indices of Type 1 and Type 2 DM.

Vital pulpotomy is a very useful method for disarming of canine tooth, tooth fracture, periodontitis, and malocclusion in veterinary dentistry. Calcium hydroxide is the material commonly used as a liner during vital pulpotomy. This creates a mineralized barrier by stimulating osteoblastic hard tissue repair, arrests the inflammatory response, and soothes dentin. However, the powder or mix type calcium hydroxide materials have many disadvantages due to complicated procedures for use and are hard to handle when vital pulpotomy is followed under general anesthesia in animals. This study was conducted in order to compare the effect of mix and premixed paste type calcium hydroxide as a liner in vital pulpotomy. Six beagle dogs underwent hemisection on the mesial root of the mandibular first molar and vital pulpotomy on the distal root of the first molar. On the distal root of the left and right mandibular first molar, mix type (DYCAL®, Dentsply, USA) and premixed paste type calcium hydroxide (VITAPEX®, Morita, Japan) were used as liners, respectively. Radiological evaluation was performed at immediate, 4, 12, and 20 weeks after vital pulpotomy. According to the results, all teeth had well-formed dentinal bridges, and there were no periradicular lucency, lamina dura loss, or anomalies of the pulp cavity. According to these results, on vital pulpotomy in animals, premixed paste type calcium hydroxide was easy to handle and decreased the anesthesia period due to a more convenient application procedure. A further study of many clinical cases is needed for evaluation of side effects and other problems.