Although Agaricus bisporus mushroom is a popular mushroom consumed world-widely, the application of common bio-elements to verify its geographical origin remains highly limited. Therefore, this study aimed to verify whether the six cultivation regions in Korea of A. bisporus could be determined by the stable isotope composition analysis of bio-elements, which are unique and abundant in most living creatures. δ13C, δ15N, δ18O, and δ34S in A. bisporus were influenced by the region, cultivar, and the interactions between these two factors (P < 0.05). In particular, the effect of cultivation region was more significant to the isotope ratio profiles as compared to the mushroom cultivar effect. During the cultivation period of A. bisporus, the C, N, O, and S isotopic fractionation was observed between the mushroom and cultivation medium, note higher in the mushroom (P < 0.05). Two dimensional plot of δ15N, δ18O, or δ34S effectively distinguished the cultivation regions, Nonsan, Buyeo, Boryung, Daegu, and/or Gyeongju examined in this study. Further, these isotope ratio profiles measured in this study would be statistically analyzed with various chemometrics to provide isotope markers for the authenticity of geographical origin. Our preliminary case study improves our understanding of how the isotope composition of A. bisporus varies with respect to cultivation regions and cultivars. In conclusion, the analysis of stable isotope ratios is a suitable potential tool for discrimination between the cultivation origins of A. bisporus collected from Korea, with potential application to other countries after certain validation steps required.

This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

Somatic cell nuclear transfer (SCNT) is the technique which generates embryos by transferring diploid nucleus into an enucleated oocyte, it has produced specific animals successfully in a variety of species. However, the developmental capacity of SCNT embryos is still relatively lower than that of embryos produced in vivo. Oocyte is a kind of lipid rich cells, its quality limits the efficiency of embryo production. L-carnitine is a co-enzyme facilitating the transportation of long chain fatty acids across the inner mitochondria membrane where fatty acids are used for generating adenosine triphosphate (ATP) via beta-oxidation. It also has antioxidant actions which may protect mitochondrial membranes and DNA against damage induced by reactive oxygen species (ROS). Whether L-carnitine is functional in bovine SCNT embryos are unknown. Therefore, the objective of this study was to examine the effects of L-carnitine on oocyte maturation and developmental competence of subsequent SCNT embryos. L-carnitine was supplemented during IVM, then intracellular ROS and GSH levels, mitochondrial activity, gene expression of COCs were analyzed at the end of IVM. SCNT embryos were produced subsequently, apoptosis detection and gene expression evaluation were performed in blastocysts. In the results, treatments with 1.5 mM and 3 mM L-carnitine significantly improved maturation rates (P<0.05). Treatments with 3 mM L-carnitine effectively induced improvement in nuclear maturation, intracellular GSH levels and mitochondrial activity, as well as a reduction in intracellular ROS levels (P<0.05). mRNA levels of CPT1A, ACAA1, ACAA2, AREG, EREG, SOD1, GPX4, GLUT1 and CDC2 transcripts were effectively up-regulated by 3 mM L-carnitine treatments (P < 0.05). Similarly, 3mM L-carnitine induced an increase in blastocyst developmental rates and an improvement in blastocyst quality (P<0.05). Our study indicates that L-carnitine treatment during IVM improves oocyte nuclear maturation and subsequent SCNT embryo development.

The mesenchymal stem cells (MSC) has been investigated as a source of stem cell therapy to replace and treat damaged cells. Human endometrial epithelial and stromal cells was isolated from hysterectomy tissue and the direct evidence of stem/progenitor cells in the human endometrium was identified. Endometrium derived stem cells (EnMSCs) are known to have a high proliferative ability, genetic stability, lack of tumorigenicity and low immnunogenicity during long-term cultivation. Here, we aimed to identify MSC in canine endometrium and characterize its potential to differentiate into decidua cells. EnMSCs were isolated from thrown-away spayed uterus of adult canine depending on their estrus cycle, and identified by flow cytometry, immunocytochemistry and flow cytometry with MSC specific markers. We then characterized the ability of EnMSCs by the doubling-time analysis, colony-forming units and MSC differentiation assays. Isolated EnMSCs expressed stem cell specific genes (Sox2, Oct4, Nanog, MCAM, Endoglin, Susd2 and IGTB) and MSC surface markers (CD90, CD44 and CD117). EnMSCs are also differentiated into adipogenic, osteogenic and chondrogenic cells morphologically under modified conditions with the expression of lineage specific genetic markers. EnMSCs showed higher proliferation ability than canine amniotic fluid derived MSCs which were used as a positive control. EnMSCs were cultured at low density (10, 20, cells/cm2) and initiated to form small colonies of loosely-arranged cells and gradually formed large colonies of densely-packed cells which underwent self-renewal with high proliferative potential which is similar to the clonogenicity feature of human endometrium-derived stem cells. EnMSCs were then induced to differentiate into decidua cells with 0.5 mM dbcAMP. After 14 days, EnMSCs changed their morphology into the elongated and rounded shape. The induced decidual cells expressed PRL and IGFBP1 which are typically expressed in decidua cells. In conclusion, we successfully isolated and characterized MSC in the canine endometrium which differentiated into decidua cells. These results showed that endometrium may be a promising source of stem cells, and furthermore raise the possibility of canine EnMSCs as a novel hypothetical decidualisation model of infertility associated with decidualisation insufficiency and implantation failure.

Activated carbons (ACs) have been used as electrode materials of electric double-layer capacitors (EDLC) due to their high specific surface areas (SSA), stability, and ecological advantages. In order to make high-energy-density ACs for EDLC, petroleum pitch (PP) precarbonized at 500–1000°C in N2 gas for 1 h was used as the electrode material of the EDLC after KOH activation. As the pre-carbonization temperature increased, the SSA, pore volume and gravimetric capacitance tended to decrease, but the crystallinity and electrode density tended to increase, showing a maximum volumetric capacitance at a medium carbonization temperature. Therefore, it was possible to control the crystalline structure, SSA, and pore structure of AC by changing the pre-carbonization temperature. Because the electrode density increased with increasing of the pre-carbonization temperature, the highest volumetric capacitance of 28.4 F/cc was obtained from the PP pre-carbonized at 700°C, exhibiting a value over 150% of that of a commercial AC (MSP-20) for EDLC. Electrochemical activation was observed from the electrodes of PP as they were pre-carbonized at high temperatures above 700°C and then activated by KOH. This process was found to have a significant effect on the specific capacitance and it was demonstrated that the higher charging voltage of EDLC was, the greater the electrochemical activation effect was.

Insect resistance to crop protecting chemicals is developing very rapidly, which is a major problem in the production of crops. Recently, the mechanism of resistance to various pseticides s has been revealed in Helicoverpa armigera. However, to date, no technology has been developed to quickly diagnose resistance yet. In this study, we present the results of developing method of rapid diagnosis of resistance

Dermanyssus gallinae is parasitic to chicken, and they cause many damages such as disturbing sleep reducing for body weight and egg production by blood-sucking. To develop acaricide against D. gallinae, The acaricidal activity of 40 species plant extract were examined. Cnidium officinale extracted by Me-OH showed 82.0% acaricidal activity after treated 48 hrs at 4,000 ppm. The hexane fraction showed 92.4% mortality against D. gallinae at 48 hrs at 2,000 ppm. Purification of the biologically active constituents from the hexane extraction with acaricidal activity was done using silica gel open column chromatography and HPLC. H1122 fraction gave 80.9% mortality to D. gallinae at 400 ppm after treated 48 hrs. H1122 fraction was analyzed by GC-MS and NMR.

It is essential to provide good environment for juvenile fish to grow and spawn in order to keep reproduction mechanism of fisheries resources. However, harvesting juvenile fish deprives adult fish of the opportunity to reproduce, thereby changing self-regulating renewable resources into non-renewable ones and preventing sustainable harvesting as well as reducing fishers’ income. We reviewed the catch status of Korea on juvenile mackerel and analyzed how reducing the catch ratio of juvenile mackerel may affect the spawning stock and Korean fishermen's revenue. In addition, we analyzed the problem of caching juvenile mackerel in Korean fisheries and current efforts to protect and reduce the catch ratio of juvenile mackerel in Korea. Furthermore, we suggested future efforts to protect juvenile mackerel. The result of the study showed that reducing the catch ratio of juvenile mackerel from 44.4% to 30.0% would increase fishers’ revenue by 60.6 billion KRW. We suggest the changing of purse seine fishery’s catching methods from night operation to day operation, relocation of fishing vessels to move to other fishing grounds when fishing vessels meet high density of juvenile mackerel, and consumers’ clever choice of consuming adult mackerel in order to accelerate the move toward protecting juvenile mackerel in Korea.

We estimate the fractal dimension of the ρ Ophiuchus Molecular Cloud Complex, associated with star forming regions. We selected a cube (v, l, b) database, obtained with J = 1−0 transition lines of 12CO and 13CO at a resolution of 22′′ using a multibeam receiver system on the 14-m telescope of the Five College Radio Astronomy Observatory. Using a code developed within IRAF, we identified slice-clouds with two threshold temperatures to estimate the fractal dimension. With threshold temperatures of 2.25 K (3σ) and 3.75 K (5σ), the fractal dimension of the target cloud is estimated to be D = 1.52–1.54, where P / AD/2 , which is larger than previous results. We suggest that the sampling rate (spatial resolution) of observed data must be an important parameter when estimating the fractal dimension, and that narrower or wider dispersion around an arbitrary fit line and the intercepts at NP = 100 should be checked whether they relate to rms noise level or characteristic structure of the target cloud. This issue could be investigated by analysing several high resolution databases with different quality (low or moderate sensitivity).

This study investigated the antidiabetic effect of amaranth grain ethanol extract (AEE) in a diabetic animal model, db/db mouse. The mice were divided into 4 groups: normal control mice (C57BL/6J), diabetic mice (C57BL/6J db/db), diabetic mice fed a lower concentration of AEE (0.3 mg/kg), and diabetic mice fed a higher concentration of AEE (0.5 mg/kg). After 10 weeks of treatment, body weights, blood insulin levels and blood glucose levels of each group were compared. At the end of treatment, the results showed that both AEE supplemented groups had lower body weights than those in the diabetic groups although higher than those in the normal groups. Moreover, in both AEE supplemented groups, serum insulin levels were higher and blood glucose levels were lower than those in the diabetic groups although both values were higher than those in the normal groups. The results of this study suggest that AEE can alleviate many of the common symptoms of diabetes in diabetic mice and, therefore, has potential as a therapeutic supplement for normalization of blood glucose and insulin levels in humans.

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.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

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.