This study was designed to evaluate the effect of bovine serum albumin (BSA) in a maturation medium on oocyte maturation and embryonic development in pigs. Immature pig oocytes were matured for 44 h in a medium supplemented with 0.4% (w/v) BSA, 0.1% (w/v) polyvinyl alcohol (PVA), or 10% (v/v) pig follicular fluid (PFF). After IVM, oocytes reached metaphase II stage were activated for parthenogenesis (PA) or used as cytoplasts for somatic cell nuclear transfer (SCNT). Nuclear maturation (89.5%, 90.7% and 91.3% for BSA, PVA and PFF, respectively) and intraoocyte glutathione contents (1.20, 1.16 and 1.00 pixels/oocyte for BSA, PVA and PFF, respectively) were not altered by the macromolecules added to maturation medium. IVM of oocytes in a medium containing BSA (21.4%) and PVA (20.7%) showed significantly lower blastocyst formation after PA than culture in medium with PFF (39.2%). After SCNT, oocytes matured in medium with BSA showed decreased embryonic development to the blastocyst stage (9.2%) compared to those matured in medium with PFF (28.9%), while 23.6% of SCNT oocytes matured in medium with PVA developed to the blastocyst stage. When the effect of BSA in a maturation medium during the first 22 h and the second 22 h of IVM in combination with PFF or PVA was examined, PVA-BSA showed a higher nuclear maturation (94.1%) than BSA-PFF (84.5%). However, there was no significant difference in the blastocyst formation among tested combinations (47.3, 52.2, 50.0, 44.4 and 49.0% for PFF-PFF, PFF-BSA, PVA-BSA, BSA-PVA and BSA-PFF, respectively). Our results demonstrate that BSA and PVA added to maturation medium can support oocyte maturation comparable to PFF-supplemented medium. However, maturation of oocytes in a BSA-containing medium decreases embryonic development after PA and SCNT when compared with the medium supplemented with PFF.

Differentiated nuclei can experimentally be returned to an undifferentiated embryonic status after nuclear transfer (NT) to unfertilized metaphase II (MII) oocytes. Nuclear reprogramming is triggered immediately after somatic cell nucleus transfer (SCNT) into recipient cytoplasm and this period is regarded as a key stage for optimizing reprogramming. In a recent study (Dai et al., 2010), use of m-carboxycinnamic acid bishydroxamide (CBHA) as a histone deacetylase inhibitor during the in vitro early culture of murine cloned embryos modifies the acetylation status of somatic nuclei and increases the developmental competence of SCNT embryos. Thus, we examined the effects of CBHA treatment on the in vitro preimplantation development of porcine SCNT embryos and on the acetylated status of histone H3K9 on cloned embryos at the zygote stage. We performed the three groups SCNT: SCNT (NT), CBHA treatment at the porcine fetus fibroblast cells (PFFs) used as donor cells prior to SCNT (CBHA-C) and CBHA treatment at the porcine SCNT embryos during the in vitro early culture after oocyte activation (CBHA-Z). The PFFs were treated with a 15 μM of CBHA (8 h) for the early culture and the porcine cloned embryos were treated with a 100 μM concentration of CBHA during the in vitro early culture (10 h). Cleavage rates and development to the blastocyst stage were assessed. No significant difference was observed the cleavage rate among the groups (82.6%, 76.4% and 82.2%, respectively). However, the development competence to the blastocyst stage was significantly increased in CBHA-Z embryos (22.7%) as compared to SCNT and CBHA-C embryos (8.6% and 4.1%)(p<0.05). Total cell numbers and viable cell numbers at the blastocyst stage of porcine SCNT embryos were increased in CBHA-Z embryos as compared to those in CBHA-C embryos (p<0.05). Signal level of histone acetylation (H3K9ac) at the zygote stage of SCNT was increased in CBHA-Z embryos as compared to SCNT and CBHA-C embryos. The results of the present study suggested that treatment with CBHA during the in vitro early culture (10 h) had significantly increased the developmental competence and histone acetylation level at the zygote stage.

Dictyophora indusiata (Vent.) Desv., “Queen of the mushroom”, is a mushroom in family Phallaceae of Basidiomycota, which is commonly used as edible and medicinal mushroom in China and Korea. This study initiated to evaluate the anti-cholinesterase, skin anti-wrinkle and melanogenesis inhibitory of 80% methanol extract from fruiting body of D.indusiata. In the anti-cholinesterase experiment, acetyl-cholinesterase and butyryl-cholinesterase inhibitory activities were performed. The extract were inhibited acetyl-cholinesterase and butyryl-cholinesterase 44.09% and 49.14% at the concentration 1 mg/mL, respectively. The skin anti-wrinkle effect of the extract were determined by measuring anti-collagenase and anti-elastase activities. While melanogenesis inhibitory activities were performed by tyrosinase, DOPA inhibitory and melanin synthesis inhibitory activities. The tyrosinase inhibitory activity of the extract were from 37.60~68.96%, while DOPA inhibitory activity were from 15.43 ~ 29.58% at the concentration ranged from 0.125~2.0 mg/mL. In addition, cellular tyrosinase activity was tested with result of the enzyme activity reduced from 99.09% to 72.91% against 25~500 μg/mL of the extract. The methanol extract of D.indusiata was inhibited melanin synthesis activity 41.86% at the concentration 500 μg/mL. The collagenase inhibitory activity of the extract from D.indusiata were 34.87%, which was comparable with the positive control, EGCG ( 45.31%). While the extract showed good inhibition of elastase enzyme (46.64%). The experiment results suggested that fruiting body of Dictyophora indusiata could be used as natural anti-cholinesterase and skin care agents.

Calocybe indica (Purkayastha, 1974), Milky mushroom, is a relatively new to the world of mushroom industry, which is belong to Lyophyllaceae of Basidiomycota, and commonly used as edible mushroom in India and Southern Asia country. This study was conducted to investigate the free radical scavenging, skin whitening and anti-collagenase activities of methanol extract from fruiting body of C.indica cultivated in Bangladesh. In addition, the polyphenol compounds of the extract were analyzed by HPLC. The mushroom extract showed good activity in lipid peroxidation which ranged from 29.35% to 55.39% at the concentration 0.125~2.0 mg/mL. The scavenging activity of the extract on 1,1-diphenyl-2-picrylhydrazy radicals were from 20.22~83.93% at the same tested concentration, which were comparable with the positive control BHT. The hydroxyl radical scavenging activity of the extract was 76.24~93.92%. The skin whitening effect of the mushroom extract was performed with UV light protecting, tyrosinase and DOPA inhibitory activities. The methanol extract absorbed UV-B wavelength with maximum level of 0.356 at 280 nm. The tyrosinase and DOPA inhibitory activities of the extract were ranged from 30.36 ~ 66.25% and 7.13 ~ 30.25% at the concentration 0.125~2.0 mg/mL, respectively. Furthermore, anti-collagenase activity were determined by measuring collagenase and elastase inhibitory activity. The collagenase and elastase inhibitory activity of the extract were 42.77% and 51.08% at the concentration 1.0 mg/mL, respectively. The experiment results suggested that fruiting body of Calocybe indica could be used as natural skin care and antioxidant agents.

In the past two decades, European ash trees (Fraxinus spp.) have been severely damaged by ash dieback disease of which causal agent is a fungal species called Hymenoscyphus fraxineus (anamorphic stage Chalara fraxinea). Recent molecular phylogenetic and population genetic studies suggested that this fungus may have been introduced from Eastern Asia to Europe. In the course of fungal biodiversity survey in Korea, H. fraxineus-like apothecia were collected from fallen leaves, rachis and petioles of Korean ash and Manchurian ash trees. The morphological and ecological traits of these materials are provided, supplemented by ITS rDNA sequence comparison of H. fraxineus strains collected from Europe, China and Japan.

Several Mites are currently the most serious threat to the world bee industry. The ectoparasitic honey bee mites was originally confined to the Asian honey bee(Apis cerana etc.). Varroa destructor and Tropilaelaps clareae has plagued European honey bees, Apis mellifera. Differences in mite tolerance are reported between two honey bee species A. mellifera and A. cerana. We were amplified antimicrobial peptide cDNA genes (Defencin, Abaecin, Royalisin, Apidaecin and Hymenoptaecin) by RT-PCR. We explored the transcriptional response to mite parasitism in A. mellifera 4th instars larvae which differ in susceptibility to V. destructor and T. clareae, comparing parasitized and non-parasitized 4th instars larvae (worker and Drone) from same hive. Differential gene expression of worker bees and Drone bees induced by mites (V. destructor and T. clareae) infection was investigated by northern blot. Mites (V. destructor and T. clareae) parasitism caused changes in the expression of genes related to sex distinction. Bees tolerant to mites (V. destructor and T. clareae) were mainly characterized by differences in the expression of genes regulating antimicrobial gene expression. It provides a first step toward better understanding molecular expression involved in this differential sex distinction host-parasite relationship. We were detected bee virus in A. mellifera, comparing parasitized and non-parasitized 4th instars larvae (worker and Drone). Therefore, this result was demonstrated that mites were another possible route of horizontal transmission, as several viruses were detected in mites and their hosts.

South Korea has over 38 millions of managed honey bee (Apis cerana) colonies before 2009 years ago, which produce the highest quantity of honey in the Korea; however, almost colony (99%) were collapsed by Korean Sacbrood Virus (KSBV) in South Korea. Korean Sacbrood Virus (KSBV) is the pathogen of A. cerana Sacbrood disease, which poses a serious threat to honeybee A. cerana, and tends to cause bee colony and even the whole apiary collapse. Colony collapse of A. cerana was first reported on the Pyeong-Chang of the South Korea in 2009. Symptoms of KSBV include the rapid transmission of larval stage honeybees (A. cerana), many dead larvae found in the bottom of hive and comb. Honeybees (A. cerana) are a very important species because they provide a number of pollination services for various ecosystems in some provinces (ex. jeon-nam, jeon-buk province). They are also extremely important organisms within human society, both agriculturally and economically. The fact that a direct cause has been determined suggests that colony collapse is a complex problem with a combination of natural and anthropogenic factors. Possible instigators of colony collapse include: wax moth, viral and fungal diseases, increased population, decreased genetic diversity, climate changing and a variety of other factors. The interaction among these potential causes may be resulting in immunity loss for honeybees and the increased likelihood of collapse.

Biological properties of antimicrobial peptides (AMPs) of hemimetabolous insect are poorly characterized in innate immunity field. To investigate the biochemical properties of hemimetabolous insect’s AMPs, we purified the pyrrhocoricin-like AMP from the hemolymph of Riptortus pedestris and then named as riptocin. We successfully determined the primary protein structure and its cDNA sequence. Interestingly, the determined cDNA revealed that riptocin precursor is composed of 12 repeating units of active riptocins, which implied that riptocin precursor might require to be processed to generate active riptocins by several unidentified processing enzymes. In order to characterize the bio-processing mechanisms of riptocin precursor, we generated the antibody against active riptocin. Using quantitative PCR and Western blot analyses, we showed that gene of riptocin was started to express from the fatbody after three hours post bacterial infection. To address our hypothesis that active riptocin is generated from riptocin precursor by several processing enzymes, we need to obtain the riptocin precursor. Currently, we are expressing the recombinant riptocin precursor using in vitro translation system. Meanwhile, we investigated whether naive hemolymph (naive HL), which may contain precursor riptocin, can generate active riptocin when riptocin precursor was co-incubation with bacteria-challenged hemolymph (active HL), which may contain all processing enzymes. Actually, when naive HL was incubated with active HL, antimicrobial activity was dramatically increased, suggesting that processing enzymes in active HL may induce processing of riptocin precursor to generate active riptocins.

Riptortus pedestris possesses Burkholderia as gut symbiont in a symbiotic organ M4 midgut. To answer why Burkholderia symbionts are not eliminated by Riptortu s immune responses, we developed two hypotheses: (i) Burkholderia symbionts do not activate host innate immunity, or (ii) Burkholderia symbionts are resistant to th e host immune responses. For the first hypothesis, we compared the antimicrobial activities of the cultured Burkholderia-injected hemolymph and symbiotic Burkhol deria-injected hemolymphs. As a result, the symbiotic Burkholderia induced antim icrobial activity like the cultured Burkholderia, indicating the symbiotic cells are st ill immunogentic to host. However, when the activated hemolymph was treated to the Burkholderia cells, the symbiotic Burkholderia showed much higher susceptibi lity than the cultured Burkholderia. To understand molecular basis of these results, we purified antimicrobial peptides (AMPs) from Riptortus hemolymph. Similarly, the symbiotic Burkholderia exhibited the high susceptibility to the purified AMPs, riptocin and rip-defensin. To understand how symbiotic Burkholderia can survive in host in spite of their immuno-susceptibility, we examined the AMP expression i n the M4 midgut. Interestingly, the expression of AMPs is suppressed in the M4 mi dgut in comparison to that of the fat body. Finally, we proposed that the immuno-su sceptibility of Burkholderia symbiont helps them to retain in the symbiotic organ. Our in vivo data showing the rapid clearance of the symbiotic Burkholderia after inj ection to host Riptortus supports our proposal.

An easy and rapid resistance detection protocol for the Western flower thrips Frankliniella occidentalis was established based on the residual contact vial bioassay (RCV), in which insecticide resistance levels can be estimated at 8 h-post treatment of diagnostic doses. The RDA strain was used as a reference susceptible strain, which has been reared under laboratory conditions over 10 years without exposure to any insecticides. Seven insecticides were tested for the determination of diagnostic dose. Among them, five insecticides (chlorfenapyr, acrinathrin, spinosad, emmamectin benzoate and thiamethoxam, ranged as 0.03 ~ 0.42 μg-1cm2) were applicable to the RCV. However, two insecticides (omethoate and imidacloprid) were not able to be used for the RCV because the treated inner surface of glass vials by these insecticides were too viscous, causing non-specific mortality. The RCV detection kit was employed for the estimation of resistance levels for the five insecticides in five local populations. Almost field-collected populations revealed high levels of resistance to the four insecticides (acrinathrin, spinosad, emmamectin benzoate and thiamethoxam) by showing less than 50% mortality. The baseline resistance detection by RCV method will facilitate the selection of proper insecticides for farmers to manage insecticide resistant-populations of F. occidentalis.

Symbiotic bacteria are common in insects. Because symbiotic bacteria are known to intimately affect the various aspects of insect host biology, ideally insects can be controlled by manipulating their symbiont. However, the attempts to control insects through their symbiont have been very limited. The paucity of the insect pest control using their symbiont is most likely due to the poor understanding of the symbiotic interactions between host insect and symbiont, which is attributed to the difficulty in cultivation of insect symbionts. However, the recently established bean bug, Riptortus pedestris, symbiotic system provides good opportunities to study insect’s symbiont in molecular level through their cultivable symbionts. Bean bugs acquire genus Burkholderia cells from environment and harbor them as their gut symbionts in the specialized posterior midgut. The genome of the Burkholderia symbiont was sequenced, and the genomic information has been used to generate the genetically manipulated Burkholderia symbiont strains. After orally administering the mutant Burkholderia symbionts into bean bugs for symbiotic association, the bacterial colonization levels in the host gut and host phenotypes were analyzed. As a result, we have identified novel symbiotic factors necessary for establishing successful association with host. Our recent understandings on the bacterial symbiotic factors demonstrate a great possibility to control the bean bug pest using genetically modified Burkholderia symbiont.

There are relatively few cases of classical biological control in Korea. During 1904 to 1905 the wooly apple aphid, Eriosoma langigerum, had been happen to invade into Korea from Japan together with imported seedling of apple and its damage spread to Jinampo near Pyoung-Yang City and it occurred all across the country by 1934. In 1934 a parasitic wasp, Aphelinus mali, was introduced and released to apple orchards and found established in nature. And it was the first case of classical biological control in Korea. Afterward couple of classical biological control programs against invaded pests were conducted mainly by the RDA with variable results. For example the orange scale, Ceroplastes rubens, and the Eggar which were successfully controlled by introduction Anicetus beneficus and Thecodiplosis pinico, respectively.

The purpose of this study is to examine the brand image and county of manufacture(COM) effect on brand luxury index, especially when the acquirer brand is afflicted by a low brand image and the acquired brand enjoys a high brand image. There were 248 responses in Taiwan (low image differences were gathered from 119 respondents and high image differences were gathered from 129 respondents). The results indicate that brand image have a positive influence on every dimension of brand luxury, while the better the luxury brand image acquired by one with an inferior image, the more the brand luxury will increase. Regarding the COM effect, the better COM image the acquirer move to, the more the brand luxury will increase. In addition, brand image and COM have interactive effect.

Sustainable marketing, as recently noticed by academics and practitioners, refers to a form of marketing that makes a net positive contribution to society in terms of environmental, social, and economic developments. Firms’ interest in sustainability as part of business performance beyond mere financial goals has been increased. Various sources, including societal mandates incorporated into regulations, concern about loss of sales, and a potential decline in corporate reputation pressures companies into implementing proper sustainability management. The purpose of this study is to clarify measures of perceived sustainability from the marketing perspective, analyze the effects of perceived sustainability on customer equity, and develop the recent theoretical frameworks and implications that will enable sustainable marketing concepts to be globally competitive. The measures of perceived sustainability items enable researchers to examine relationships among perceptions of sustainability and other key customer equity variables, such as value equity, brand equity, and relationship equity. For this and other reasons, the MPS may have value to practitioners. By understanding perceived sustainability, they can develop economic, social, environmental and communication performances that effectively utilize sustainability. The measures of perceived sustainability offers researchers a tool for measuring perceived sustainability that is consistent with the literature on sustainability, while recognizing the reality that sustainability is a multidimensional construct. The rigor reflected in the multiple methods for generating scale items and the multiple stages in the scale development process results in a scale that should be useful to both researchers and practitioners.

Russula compacta, a wild mushroom, belongs to Russulaceae, Russulales of Basidiomycota. This study was conducted to evaluate the free radical scavenging, anti-inflammatory, anticholinesterase and anti-α-glucosidase effects from fruiting bodies of R. compacta extracted with methanol and hot water. In 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging effects, the methanol and hot water extracts showed good scavenging effects comparable with positive control, BHT. The chelating effect of methanol and hot water extracts of the mushroom were significantly higher than the positive control, BHT. The reducing power of the methanol and hot water extracts of the mushroom were lower than the positive control at the concentrations tested. In the HPLC anaysis of phenolic acids profile of the mushroom extract, 7 phenolic acids such as gallic acid, vanillin, rutin hydrate, resveratol, quercetin formononetin, and biochanin-A were detected. Nitric oxide (NO) production in lipopolysaccahride (LPS) activated RAW 264.7 cells was inhibited by 1.5-fold with the treatment of methanol extract when compared with the control. In the anti-cholinesterase activity assay, the methanol extract inhibited the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) effects by 73.9% and 81.05% at the 1.0 mg/mL concentration, whereas galanthamine, the standard drug, inhibited the AChE and BChE activities by 97.80% and 81.12%, respectively at the same concentration. The methanol and hot water extracts of the mushroom inhibited the α-glucosidase activity by 55.44% and 62.00%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control inhibited the α-glucosidase activity by 81.81% at the 2.0 mg/mL concentration. From the experimental results, the fruiting bodies of R. compacta contained natural antioxidant, anti-inflammatory, anti-cholinesterase, and anti-diabetic substances, which might be used for health foods.

Coprinellus miaceus, belongs to Coprinaceae of Agaricales, Basidiomycota, has been used for an edible purposes in asian countries. This experiment was initiated to evaluate the free radical scavenging, free radical scavenging, anti-acetylcholinesterase, anti-inflammatory and α-glucosidase inhibitory activities of C. micaceus fruiting bodies extracted with methanol and hot water. In 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, the scavenging activities of methanol and hot water extracts were lower than that of positive control, BHT. The chelating effects of methanol and hot water extracts were significantly higher than positive control, BHT at the concentrations of 0.125-2.0 mg/mL. In the reducing power assay, methanol and hot water extracts exhibited the lower activities than the positive control, BHT at the 0.125-0.2 mg/mL concentration. In the HPLC analysis of phenolic acids profile of the mushroom fruiting bodies, 4 phenolic compounds including procatechuic acid, chlorogenic acid, (-)-epicatechin, and naringin were detected. The tyrosinase inhibitory activities of methanol and hot water extracts were 91.33% and 91.99% at 2.0 mg/mL concentration, while the inhibitory activity of kojic acid, the positive control, was 99.61%. Nitric oxide (NO) production in lipopolysaccahride (LPS) activated RAW 264.7 cells were inhibited by the methanol extract in a concentration dependent manner. In the acetylcholinesterase (AChE) inhibitory activity assay, methanol and hot water extracts of the mushroom inhibited the AChE by 94.64% and 74.19%, respectively at the 1.0 mg/mL concentration, whereas galanthamine, the standard drug inhibited the AChE activity by 97.80% at the same concentration. The methanol and hot water extracts of the mushroom inhibited the α-glucosidase activity by 62.26% and 67.59%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control inhibited the α-glucosidase activity by 81.81% at the same concentration. Therefore, it is concluded that fruiting bodies of C. micaceus contained natural antioxidant, anti-inflammatory, anti-acetylcholinesterase and α-glucosidase inhibitory substances which might be used for promoting human health.

Lentinus giganteus is a edible mushroom cultivated in Asian countries. The present study was initiated to evaluate the anti-inflammatory, anti-acetylcholinesterase, anti-α-glucosidase, and free radical scavenging activities from fruiting bodies of L. giganteus extracted with methanol and hot water. The free radical scavenging activities of methanol and hot water extracts on 1,1-diphenyl-2-picrylhydrazyl (DPPH) were 92.26% and 90.17% at 2.0 mg/mL concentration, respectively and comparable with positive control, BHT. The chelating activities of methanol and hot water extracts were significantly higher than the positive control tested. The reducing power of methanol and hot water extracts showed lower activities compared to positive control, BHT. The phenolic and flavonoid contents of hot water extract were 1.56 μg/mg and 24.35 μg/mg, respectively. Nitric oxide (NO) produced by lipopolysaccahride (LPS) activated RAW 264.7 cells were significantly inhibited by treatment of methanol and hot water extracts. The methanol extract inhibited the acetylcholinesterase (AChE) activity by 91.19% at the 2.0 mg/mL concentration, whereas galanthamine, the standard drug, inhibited the AChE activity by 97.80%. The hot water extracts inhibited the α-amylase and α-glucosidase activities by 78.86% and 80.78%, respectively at the 2.0 mg/mL concentration, while acarbose, the positive control, inhibited the activities by 89.91% and 81.81%, respectively at the same concentration. Therefore, it is concluded that fruiting bodies of L. giganteus contain antioxidant, anti-inflammatory, anti-cholinesterase, and anti-diabetic substances, which can be used for natural health food for promoting human health.