Royal jelly (RJ) is a well-known functional and medicinal food for human health promotion. Major royal jelly proteins (MRJPs), which are the major protein components in RJ, exhibit antimicrobial activities. However, the identities of the MRJPs of RJ responsible for its antioxidant effects have remained unclear. Here, we report that honeybee (Apis cerana) MRJP 2 (AcMRJP2) acts as an antimicrobial and antioxidant agent in RJ. Using recombinant AcMRJP2, which was produced in baculovirus-infected insect cells, we established the antimicrobial and antioxidant roles of MRJP 2. AcMRJP2 bound to the surfaces of bacteria, fungi, and yeast, which then induced structural damage in the microbial cell walls and led to a broad spectrum of antimicrobial activities. AcMRJP2 protected mammalian and insect cells via the direct shielding of the cell against oxidative stress, which led to reduced levels of caspase-3 activity and oxidative stress-induced cell apoptosis, followed by increased cell viability. Moreover, AcMRJP2 exhibited DNA protection activity against reactive oxygen species (ROS). Our data indicate that AcMRJP2 could play a crucial role as an antimicrobial agent and antioxidant in RJ, suggesting that MRJP 2 is a component responsible for the antimicrobial and antioxidant activities of RJ.

Bee venom contains a variety of peptide constituents, including low-molecular-weight protease inhibitors. While the putativelow-molecular-weight serine protease inhibitor Api m 6 containing a trypsin inhibitor-like cysteine-rich domain was identifiedfrom honeybee (Apis mellifera) venom, no anti-fibrinolytic or anti-microbial roles for this inhibitor have been elucidated.In this study, we identified an Asiatic honeybee (A. cerana) venom serine protease inhibitor (AcVSPI) that was shownto act as a microbial serine protease inhibitor and plasmin inhibitor. AcVSPI was found to consist of a trypsin inhibitor-likedomain that displays ten cysteine residues. Interestingly, the AcVSPI peptide sequence exhibited high similarity to the putativelow-molecular-weight serine protease inhibitor Api m 6, which suggests that AcVSPI is an allergen Api m 6-like peptide.Recombinant AcVSPI was expressed in baculovirus-infected insect cells, and it demonstrated inhibitory activity against trypsin,but not chymotrypsin. Additionally, AcVSPI has inhibitory effects against plasmin and microbial serine proteases; however,it does not have any detectable inhibitory effects on thrombin or elastase. Consistent with these inhibitory effects, AcVSPIinhibited the plasmin-mediated degradation of fibrin to fibrin degradation products. AcVSPI also bound to bacterial andfungal surfaces and exhibited anti-microbial activity against fungi as well as gram-positive and gram-negative bacteria. Thesefindings demonstrate the anti-fibrinolytic and anti-microbial roles of AcVSPI as a serine protease inhibitor.

Introduction E-retailers take advantage of apparel products which are viewed as a constantly changing experiential product rich that lead to various kinds of hedonic consumption, like impulse buying (Park and Kim, 2008). Apparel purchases are often affected by irrational and emotional attraction making them one of the most frequent impulsively purchased items online (Rhee, 2007). The e-shoppers are more spontaneous due to marketing stimuli which makes it easier to purchase impulsively and reduces risk aversion (Madhavaram and Laverie, 2004). In fact, shoppers are more likely to overspend when e-shopping because it does not feel like spending money (Dittmar, Long, and Meek, 2004). Apparel retailers need to pay special attention to converting web browsers to impulse purchasers—a capability that plays an important role in the growth of e-business. Despite the vast amount of data available online, few efforts have been made to identify the relationship among services attributes, browsing, urges to buy, and e-impulse buying of apparel products. This study explores a model of e-impulse buying for strategic e-business management in apparel products by understanding critical factors of e-store service attributes over the shopping websites and its impact on browsing and urges to buy for apparel in Korea. Specifically, the objectives of this study were to (a) identify underlying factors of online service attributes related to apparel products; (b) estimate structural equation model for causative relationship among service attributes, browsing, urges to buy, and e-impulse buying; and (c) examine mediating effect of browsing and urges to buy on e-impulse buying for apparel. By focusing on apparel products, this study will shed light on complex issues surrounding online service and provide opportunities for strategic development and promotion in fashion merchandising online. Also, this study could help mangers to identify successful global applications of e-marketing for apparel products. Methods A self-administered questionnaire was developed based on previous literatures. The instrument consisted of the main variables; e-service attributes of apparel stores, browsing, urge to buy, and e-impulse buying of apparel products. Participants responded to questions on a 5-point rating scale (1=very unlikely to 5=very likely). The questionnaire was administered during a regularly scheduled class in universities. Usable data were obtained from 319 students aged between 18 and 25 residing at the metropolitan areas in Korea. More than half of the respondents (65.5%) were 18 to 21 years old. Nearly 65.5 percent of the subjects were women, and more than half of the respondents (63.8%) were below the sophomore. For data analysis, exploratory factor analysis with varimax rotation was conducted to determine dimensions of perceived service attributes of the apparel online store. Cronbach's alpha established inter-item reliability between items. The structural equation model analysis was conducted by AMOS 18.0 using a correlation matrix with maximum likelihood approach. The overall fit of the model was assessed by various statistic indexes: chi-square (χ2), goodness of fit index (GFI), adjusted goodness of fit index (AGFI), and root mean squared residual (RMR). Results An exploratory factor analysis revealed three types of e-store service attributes: Ease of Use, Accuracy, and Reliability. The structural equation model, which was estimated to examine causal relationships among variables (i.e. three types of service attributes, browsing, urges to buy, and e-impulse buying), was relatively acceptable (chi-square value = 168.23, df = 157, p = .026; GFI = .95, AGFI = .93, RMR = .05). The model demonstrated that the two factors of service attributes (i.e. ease of use, accuracy of service) were significant variables to influence the browsing, which influence the urges to buy and e-impulse buying for apparel stores. E-impulse buying of apparel products was influenced by browsing and urges to buy. In addition, the urges to buy and browsing mediated the casual relationship between service attributes and e-impulse buying for apparel products. This result suggests that browsing and urges to buy perceived by shoppers were important predicting e-impulse buying of apparel products. Especially, urges to buy apparel products was the most important mediator to trigger e-impulse buying of apparels. Conclusions & Implication This study explores a structural equation model for understanding e-impulse buying in conjunction with browsing, urges to buy and service attributes of apparel store perceived by shoppers in Korea. The findings suggest that the browsing and urges to buy perceived by shoppers play important roles as mediators of the e-impulse buying for apparel stores. Especially, service attributes of apparel e-store perceived by shoppers had indirect effects on e-impulse buying through the browsing and urges to buy, implying that the consumers perceived the ease to use of e-store was the more browsing at e-store and then the more perceiving the urges to buy apparel products. However, the more browsing at apparel e-store was the less the e-impulse buying of apparel products and the more perceiving urges to buy at e-store is the more e-impulse buying of apparel products. This study provides retail managerial implications for stimulating e-shoppers' impulse buying of apparel products. They could increase shoppers' unplanned purchases by developing the service program for convenience (e.g. easy to use the structure of contents, wide ranges of products/service packages, various menu options of website) to stimulate the browsing at e-store.

Insect cuticular melanization is regulated by the prophenoloxidase (proPO)- activating system, which is also involved in the innate immune reaction. Here, we demonstrate how the differentiation of the proPO-activating system is regulated toward a cuticular melanization or innate immunity function in silkworm (Bombyx mori) pupae. Our results indicate that the differential and spatial regulation of key components, such as the proPO-activating factor, tyrosine hydroxylase, and porPOs, primes the proPO-activating system for either cuticular melanization or innate immunity. This dual strategy for cuticular melanization in development and innate immunity upon infection demonstrates a two-pronged defense mechanism that is mediated by the priming of the proPO system.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.

Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

Bee venom contains a variety of peptides and enzymes, including serine proteases. Here we describe the molecular cloning and characterization of a serine protease (Bt-VSP) isolated from the venom of the bumblebee Bombus terrestris. The Bt-VSP gene consists of six exons encoding a 358-amino acid protein. The form of Bt-VSP detected in bee venom was the 34-kDa mature protein, which is created by cleavage of the catalytic domain of Bt-proVSP between Arg111 and Val112. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. The finding that Bt-VSP acts as a fibrin(ogen)olytic enzyme is similar to a previous finding that Bi-VSP, a venom serine protease of B. ignitus, exhibits fibrin(ogen)olytic activity. We also compared major venom components in honeybee and bumblebee, and found that bumblebee venom contains a larger amount of serine protease. Furthermore, unlike bumblebee venom, which exhibits fibrin(ogen)olytic activity owing to the presence of a serine protease, it is likely that honeybee venom lacks fibrin(ogen)olytic activity.

We cloned and characterized two peroxiredoxins (Prxs), BiPrx1 (a 1-Cys Prx) and BiTPx1 (a 2-Cys Prx) from the bumblebee Bombus ignitus. The BiPrx1 gene consists of 5 exons, encoding 220 amino acid residues with one conserved cysteine residue. The BiTPx1 gene consists of three exons, encoding 195 amino acid residues with 2 conserved cysteine residues. Recombinant BiPrx1 (27 kDa) and BiTPx1 (25 kDa), expressed in baculovirus-infected insect Sf9 cells, reduced H2O2 in the presence of electrons donated by dithiothreitol. Unlike BiTPx1, however, BiPrx1 did not show reduction activity when thioredoxin was used as the electron donor. Both BiPrx1 and BiTPx1 protected super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Tissue distribution analyses showed the presence of BiPrx1 and BiTPx1 in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting that BiPrx1 and BiTPx1 are not secretable. When H2O2 was injected into B. ignitus bees, BiPrx1 and BiTPx1 transcripts were acutely up-regulated in the fat body tissues. We also demonstrated regulation of BiPrx1 and BiTPx1 expression via reduction of transcript levels in the fat body with RNA interference (RNAi). Under H2O2 overload, the RNAi-induced BiPrx1 knock-down B. ignitus worker bees showed up-regulated expression of BiTPx1. Reciprocally, BiTPx1 RNAi knockdowns showed up-regulated BiPrx1 expression in the fat body. These results indicate that loss of expression of BiPrx1 or BiTPx1 is compensated by up-regulation of expression of the other peroxidase in response to H2O2 overload.

To study the signaling effect of insulin-like growth factor-Ⅰ(IGF-1), transgenic mice containing IGF-1 Receptor (IGF-1R) cDNA fused to metallothionein promoter were produced by DNA microinjection into the pronucleus of mouse zygote. Three founders were produced with transgenic mice containing IGF-1R gene. Transgenic mice lines contained approximately 4～20 copies of transgenes per cell and transmission of this gene into the progeny with Mendelian manner were determined. The founder mice were mated with normal mice to produce F1 mice and then F2 mice. Transmission rates of IGF-1R transgene in the progeny mice were 25～60% in F1 generation and 40～50% in F2 generation. The mRNA expression of IGF-1R transgene in liver was analyzed using RT-PCR for IGF-1R gene in liver. When body weights of transgenic pups were measured during 4, 10 and 14 weeks after birth, IGF-1R transgenic mice grew faster than non transgenic littermates. This study indicated that growth regulation by IGF-1 signaling through IGF-1R can be elucidated using IGF-1R transgenic mice.

본 연구는 LPS로 유도된 RAW 264.7 세포에서 다양한 약용식품으로 사용되고 있는 올리브 잎과 가지 추출물의 항염증 효과를 확인하였다. 올리브 잎과 가지 추출물은 각각 RAW 264.7 세포에 대하여 세포독성을 나타내지 않았고, LPS 자극에 의한 NO 및 PGE2 생성을 농도 의존적으로 억제했다. 또한, 올리브 추출물은 LPS 자극으로 분비된 TNF-α, IL-1β 및 IL-6의 전염증성 cytokine의 분비량을 억제하였으며, 특히 200 μg/mL 농도에서 올리브 가지 추출물이 잎 추출물 보다 IL-6를 억제하는 것으로 나타났다. 대표적인 염증 관련 신호 전달 경로 인자인 iNOS 및 COX-2의 발현을 검토한 결과 올리브 추출물은 iNOS의 발현을 농도의존적으로 현저히 감소시키는 것으로 관찰되었으나, 각각의 올리브 추출물이 COX-2 발현에는 영향을 미치지 않은 것으로 관찰되었다. 이와 같은 결과는 올리브 각 부위별 추출물은 모두 iNOS 및 NO 조절 경로를 조절하는 것으로 사료되나 iNOS 및 COX-2 단백질 발현은 병립적이지 않을 수 있음을 제시하고 있다. 본 연구 결과로 올리브 추출물이 독성과 부작용이 적은 항염증 효능을 가진 기능성 화장품 소재로써 개발 가능성이 있다고 사료된다.

본 연구는 장애전담 어린이집에 재원 중인 발달지체유아를 대상으로 요일별 신체리듬을 고려한 원예활동 프로그램이 유아의 적응행동에 미치는 영향에 대해 알아보고자 하였다. 원예활동 대상자는 S장애전담 어린이집에서 4세~6세 발달지체유아 대조군 8명, 실험군 8명이었다. 원예활동 프로그램은 2012년 10월 15일부터 2012년 11월 22일까지 주 3회 30분씩 총 18회 실시하였다. 전체 적응행동 평가 결과 대조군의 경우 실험 전 65.00점에서 실험 후 66.00점으로 1.00점이 향상되었고 실험군은 전체 적응행동 평가 결과 실험 전 66.00점에서 실험 후 85.50점으로 19.50점이 향상되었다. 실험군의 하위항목별 변화를 살펴보면 개념적 적응행동은 원예활동 실시 전 64.00점에서 실시 후 80.75점으로, 사회적 적응행동은 65.50점에서 80.63점으로, 실제적 적응행동은 74.38점에서 95.38점으로 모든 항목이 원예활동 실시 후 유의하게 향상되는 것을 볼 수 있었다. 이상의 연구결과로부터 요일별 신체리듬의 변화와 유아의 흥미와 참여도를 고려하여 설계한 원예활동프로그램이 장애전담 어린이집 내에 재원하고 있는 발달지체유아들의 적응행동 향상에 효과적인 것으로 나타났다.

“Jokyoung”, a new bread making wheat cultivar, was developed from the cross between “Seri 82”, a hard white wheat from CIMMYT, Mexico and “Keumkang”, a hard white wheat with high milling rate and early maturing from Korea by National Institute of Crop Sci