Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

The mammary gland is a complex organ, made up of various cell types that work together for mil synthesis. A previous study had established a clonal cell line from primary bovine mammary alveolar cells (MAC-T) for the study of bovine milk production and synthesis. In this study, we transplanted MAC-T bovine alveolar cell line to BALB/C nude mice for regeneration of bovine mammary gland alveolar ducts. The MAC-T cells were suspended with matrigel and injected into the dorsal tissue of 8 weeks old male BALB/C nude mice. After 6 weeks, the injected cells were showed typical morphology of the tubuloalveolar female glands by histological analysis. It was made the branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14 and CK18 positive cells but not in the non-transplanted MAC-T cells. These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14 and CK18 positive MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

The sweet potato whitefly, Bemisia tabaci (Gennadius) and the western flower thrips, Frankliniella occidentalis (Pergande) are major insect pests that causes crop damage worldwide by piercing leaves, sucking sap and transmitting numerous plant viruses. A new strategy for IPM, the push–pull method uses a combination of repellent intercrops (push) and alluring trap plants (pull) to manipulate the distribution of insect pests and control their populations. So, we surveyed the responses of these pests of tomato to several plants in green house. Lavandula angustifolia, Petunia hybrid, Ocimum basilicum and Rosmarinus officinalis showed about forty-percent push response to F. occidentalis in tomato. However, Gypsophila paniculata attracted the F. occidentalis in tomato on the contrary. Pelargonium tomentosum showed about fifty-percent push response to B. tabaci in tomato. However, Mentha spicata and Gypsophila paniculata attracted the B. tabaci in tomato. The utilization technique of these plants should be more inspected in further study.

Bacillus thuringiensis (Bt) produces a variety of insecticidal crystal proteins and widely used as one of the most successful biological control agents. Recently, studies that introduce cry genes into crops to create pest resistance have made much progress, and the total area of land planted with Bt crops has increased substantially. In this study, pest resistance of 8 transgenic Bt rice events with a synthetic cry1Ac gene linked to rice rbcS-tp sequence were assessed under laboratory conditions. Bioassays were performed against Cnaphalocrocis medinalis, which is a significant pest of rice in Asia. C. medinalis larvae were shown to be susceptible to all eight events, even though there were differences between the causes of death. The results differed between developmental stages of the larvae, despite the fact that all 8 events led to high mortalities. These results may be a significant foundation for the evaluation of improved transgenic Bt rice.

Cotton aphid, Aphis gossypii Gloever is one of the major pests on a wide range of economically important crops in the world. The sustained use of chemical insecticides to control the aphid has led to the emergence of resistant strains to numerous used insecticides. As an alternative strategy entomopathogenic fungi have been used as part of integrated pest management program to control aphid, especially insecticide-resistance population. In particular, Beauveria bassiana-based commercial bio-insecticide has been used to reduce the pest population under greenhouse conditions in various countries. In this study, we investigated the control efficacy of a prototype of commercial mycopesticide using an B. bassiana (wettable powder) against cotton aphid on potted cucumber plant in greenhouse conditions.

The habitat of the cockroach varies by species. The German cockroach (Blattella germanica) lives in human dwellings, while the Japanese field roach (Blattella nipponica) lives in a mountainous region. Based on phylogenetic analysis of mtCO I, the two species are closely related to each other and B. germanica is divergent from wild species such as B. nipponica. Their habitats and walking speed differ even though the two species have similar morphology. We hypothesized that habitats might influence walking speed via changes to appendage morphology and enzyme-based physiological differences. We found that phenotype such as appendage length and esterase isozyme expression were clearly different between the two species. These differences might be responsible for the observed difference in walking speed.

In this study, we investigated the effects of different pre-treatment conditions such as blanching and drying process to improve its quality. Bracken samples were treated by drying (70°C for 40 min), blanching (100°C for 2 min) or mixed (blanched and dried [BD]), respectively. These treated samples were analyzed for their physicochemical properties such as pH, color and microbial growth. The pH of bracken increased from 6.6 to above 6.9 through all treatment. From color observation, the L* and b* values increased after drying process, whereas, the a* values decreased. Water contents of bracken decreased by about 80% from 93% through drying process. After samples treated by pre-treatments, hardness increased, especially after drying process. For the microbial study, raw bracken had 5.6 log CFU/g of aerobic bacteria and 2.8 log CFU/g of total coliform. Blanched and BD samples had about 2 log CFU/g of aerobic bacteria and 1.8 log CFU/g of total coliform, acceptable for food. From our results, it is concluded that the properties of blanched samples had similar to raw samples to guarantee for microbial safety. From the obtained results, the blanching process without drying process is necessary to apply freezing process as pretreatments.

Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.

Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.

Populus euramericana is emerging as a viable feedstock for producing bioethanol from renewable resources. Steam explosion pretreatment of P. euramericana can solubilize a significant portion of the hemicellulosic component and enhance the glucose conversion of the remaining cellulose for fermentation into ethanol. In this study, steam explosion condition of P. euramericana is performed in a steam explosion reactor at severity log Ro 4.02 and severity log Ro 4.37. Glucose conversion varied from 72.3% to 80.1% of steam exploded P. euramericana at severity log Ro 4.02 and severity log Ro 4.37. Ethanol yields(%) based on sugar content after enzymatic hydrolysis after 48 h fermentation ranged from 87.0% to 88.4%. As a result, from 100 g of raw material, 14.0 g of ethanol are recovered of 47.3 g available cellulose content. This research of steam explosion pretreatment was a promising method to improve glucose conversion and ethanol yield for bioethanol production.

To obtain information on the sanitary of indoor environment in greenhouses used for shiitake cultivation, bacteria associated with larvae and imagoes of Sciaridae flies, pest to shiitake mushroom, were isolated and identified. A total of 1,048 bacterial colonies were isolated from the flies’ larvae and 984 bacterial colonies were isolated from the flies’ imagoes. Based on molecular analysis of the 16S rDNA sequence, Achromobacter xylosoxidans and Tetrathiobacter kashmirensis of β-proteobacteria, Enterobacter asburiae and Raoultella ornithinolytica of γ -proteobacteria, Curtobacterium sp. and Microbacterium thalassium of Actinobacteria, and Penibacillus taichungensis of Firmicutes were identified from the colonies of the flies’ larvae. While, Bacillus megaterium, B. thuringiensis, Lysinbacillus sphaericus and L. fusifomis of Firmicutes, Microbacterium thalassium and Citricoccus parietis of Actinobacteria, and Enterobacter asburiae of γ-proteobacteria were identified from the flies’ imagoes. Some of the isolated bacterial species were known be human pathogens. Overall, the results of this study suggested that mushroom fly carrying human pathogenic bacteria is one of sources impact on the sanitary of indoor environment of greenhouses used for shiitake cultivation.

Human mouth environment is known to include a variety bacteria, including Streptococcus spp., Staphylococcus spp., Actinomyces spp., Lactobacillus spp., Candida spp., Enterobacteriaceae, et al. Human oral microorganisms can cause dental caries, gingivitis, periodontitis, respiratory tract infection, and cardiovascular disease. Thus, right denture cleaning is essential to oral and general human health. The aim of this study was to evaluate the bactericidal effect of a sodium dichloroisocyanurate-based effervescent tablet (Aos Denti Germ, Aos Company, Chungbuk, Korea) against oral microorganisms. A total of 5 species Streptococcus spp. (Streptococcus anginosus, Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, and Streptococcus sobrinus), Actinomyces oris, Candida albicans, and Escherichia coli were used in this study. All strains were exposed to the distilled water prepared with effervescent tablet. After the exposure, the mixture of strains and effervescent tablet was inoculated onto blood agar or MacConkey agar plate and cultured at 36℃. All strains were killed immediately on exposure to effervescent tablet. The results suggested that effervescent tablet could be used as an effective denture cleanser for dental hygiene.