식품첨가물로 사용되는 알긴산나트륨은 알긴산염류로서 안정제, 증점제, 유화제 등의 기능을 한다. 알긴산나트륨의 정량법은 전처리가 복잡하고 분석시간이 많이 소요되어 상대적으로 간편하고 보편적인 분석법 연구가 요구되고 있 다. 분석장비로는 HPLC-UVD 및 Unison US-Phenyl 컬럼을 사용하였으며, 전처리 조건으로 진탕기를 이용하여 실온에서 150 rpm으로 180분간 추출하였다. 알긴산나트륨의 표준 용액을 5개 농도 범위에서 검량선을 작성한 결과 직선성(R2)은 평균 0.9999로 측정되었으며 검출한계(LOD) 및 정량한계(LOQ)는 각각 3.96 mg/kg, 12.0 mg/kg이었다. 또한, 천사채를 이용해 얻은 일내 및 일간 평균 회수율과 정밀도는 각각 98.47-103.74%, 1.69-3.08 RSD%이고, 빙과류에 대한 일 내 및 일간 평균 회수율과 정밀도는 각각 99.95-105.76%, 0.59-3.63 RSD%이다. 상대불확도%는 CODEX의 기준에 적합한 1.5-7.9%의 결과를 나타냈다. 본 연구에서 확립한 방법의 적용성 검토를 위해 총 103개 품목에 대한 알긴산나트륨의 함량을 정량한 결과 당면, 유탕면, 당류가공품 유형 순으로 높은 검출율을 보였다.

Sestrin-2 (SESN2) as a stress-metabolic protein is known for its anti-oxidative effects as a downstream factor of PERK pathways in mammalian cells. However, the expression patterns of SESN2 in conjunction with the UPR signaling against to ER stress on porcine oocyte maturation in vitro, have not been reported. Therefore, we confirmed the expression pattern of SESN2 protein, for which to examine the relationship between PERK signaling and SESN2 in porcine oocyte during IVM. We investigated the SESN2 expression patterns using Western blot analysis in denuded oocytes (DOs), cumulus cells (CCs), and cumulus-oocyte complexes (COCs) at 22 and 44 h of IVM. As expected, the SESN2 protein level significantly increased (p < 0.01) in porcine COCs during 44 h of IVM. We investigated the meiotic maturation after applying ER stress inhibitor in various concentration (50, 100 and 200 μM) of tauroursodeoxycholic acid (TUDCA). We confirmed significant increase (p < 0.05) of meiotic maturation rate in TUDCA 200 μM treated COCs for 44 h of IVM. Finally, we confirmed the protein level of SESN2 and meiotic maturation via regulating ER-stress by only tunicamycin (Tm), only TUDCA, and Tm + TUDCA treatment in porcine COCs. As a result, treatment of the TUDCA following Tm pre-treatment reduced SESN2 protein level in porcine COCs. In addition, SESN2 protein level significantly reduced in only TUDCA treated porcine COCs. Our results suggest that the SESN2 expression is related to the stress mediator response to ER stress through the PERK signaling pathways in porcine oocyte maturation.

The national natural monument of Korea, Jeju Black Cattle (JBC), it is a native species with unique blood line. This cattle breed needs mass production and industrialization to further improve and preserve their characteristics. This study was to examine whether there were differences in in vitro developmental rates according to body weight (<300, 300 ~ 350, 350 ~ 400 and >400 kg) and grade (1++, 1+, 1, 2 and 3), and oocyte donors or non-donors. As a method of IVM, groups of ten cumulus oocyte complexes (COCs) were cultured in 50 μl droplets of maturation medium (TCM199 supplemented with 10% FBS, 0.2 mM sodium pyruvate, 1 μg/ml follicle-stimulating hormone, 1 μg/ml estradiol-17β) under mineral oil at 38.8℃ in an incubator with a 5% CO2 atmosphere for 22 to 24 h. For IVF, 44 ul IVF drop contained 10 oocytes with sperm concentration of 1 × 106 cells/ml, and then 2 μl heparin and 2 μl PHE (20 μM peicillamine, 10 μM hypotaurine, 2 μM epinephrine) were added. For IVC, after 44±2 h of incubation, cleaved embryos were incubated in CR1aa medium containing 3 mg/ml FAF-BSA until day 4 at 38.8℃ in a 5% CO2 incubator. Embryos were then cultured in CR1aa medium containing 10% FBS until day 8. As a result, in vitro development rates were the highest in 350 ~ 400 kg body weight group and in 1++ grade group than other groups (p<0.05). However, there was no difference in in vitro developmental capacity of classified donor and non-donor oocyte groups. This result demonstrated that the better in vitro developmental capacity was obtained in high level originated oocyte groups (350 ~ 400kg, 1++ grade) than in others, while there was no different in donor types.

구상나무는 아고산대에 한정되어 생육하는 것으로 알려져 있는데 최근 국립공원 내에서 고사하는 개채가 많이 발견되고 개체 수 또한 줄어들고 있는 실정이다. 따라서 본 연구는 쇠퇴 현상을 보이고 있는 구상나무 군락의 토양 환경 특성 파악을 통하여 추후 국립공원 내 구상나무 군락의 보전방안 마련 및 관리를 위한 기초자료를 제공하고자 수행되었다. 국립공원 내 구상나무군락의 토양 환경 특성을 파악하기 위하여 소백산 비록봉(1지점), 덕유산 서봉(1지점), 향적봉(4지점) 그리고 지리산 노루목(1지점), 돼지평전(1지점), 반야봉(2지점), 벽소령(1지점), 세석평전(1지점), 영신봉(1지 점), 임걸령(1지점), 장터목(1지점), 제석봉(2지점)에 존재 하는 구상나무 군락을 조사 대상지로 선정하였다. 토양의 물리적 특성을 분석한 결과 평균 토심은 소백산에서 68cm, 덕유산에서 41cm 그리고 지리산에서 44cm로 나타났으며, 특히 지리산 돼지평전과 벽소령 지역 내 구상나무군락의 경우에는 10cm 이하로 매우 낮게 조사되어 매우 열악한 토양 환경을 보이고 있었다. 0~10cm 깊이에서 가비중 평균은 소백산에서 0.38g/㎤, 덕유산에서 0.48g/㎤, 지리산에서 0.62g/㎤로 분석되어 우리나라 산림토양(정진현 등, 2002) A층 평균 0.88g/㎤ 그리고 B층 평균 1.01g/㎤보다 낮은 수준을 보이고 있었으며, 토심이 깊어질수록 가비중은 증가하는 경향을 보이고 있었다. 토양 수분함량의 경우에는 0~10cm 깊이에서 평균 39.69%~47.57%로 나타났다. 토양의 화학적 특성을 분석한 결과 토양 산도(pH)는 소백산에서 평균 4.90, 덕유산에서 4.59 그리고 지리산에서 4.69로 우리나라 산림의 토양 산도(pH) A층 평균 5.48 그리고 B층 평균 5.52보다 매우 낮게 나타났다. 토양 내 유기물 함량은 소백산에서 평균 9.08%, 덕유산에서 12.92% 그리고 지리산에서 12.68%로 양호한 수준이었으며, 모든 조사 지점에서 우리나라 산림토양 내 A층 유기물 평균 4.49%, B층 평균 2.03% 보다도 높은 함량을 보이고 있었다. Miller and Donahue(1990)에 의하면 유기물은 토양 중 양이온 치환용량(C.E.C.)의 30~70%를 제 공하며 부식으로 인하여 양이온치환 입지가 제공되는 것으로 알려져 있는데, 본 연구대상지의 양이온 치환용량을 분석한 결과 유기물 함량이 상대적으로 높았던 덕유산과 지리산에서 각각 22.05cmolc･kg-1과 19.69cmolc･kg-1로 높게 나타났다. 그리고 유기물 함량이 상대적으로 낮았던 소백산에서 17.58cmolc･kg-1로 조사되었는데, 이는 우리나라 산림토양 내 A층 양이온 치환용량 평균 12.5cmolc･kg-1, B층 평균 10.7cmolc･kg-1 보다는 높은 수준을 보이고 있었다. 유기물과 밀접한 관계를 가지고 있는 전질소 함량의 경우에도 소백 산에서 평균 0.52%, 덕유산에서 0.58% 그리고 지리산에서 0.59%로 조사되어 우리나라 산림토양 내 A층 전질소 평균 0.19%, B층 평균 0.09% 보다도 높은 함량을 보이고 있었다. 치환성양이온 K+은 덕유산과 지리산 조사 지역에서 평균 0.20cmolc･kg-1로 조사되어 우리나라 산림토양 A층 평균 0.22cmolc･kg-1, B층 평균 0.21cmolc･kg-1과 유사한 수치를 보였으나, 소백산 구상나무림의 경우에는 0.10cmolc･kg-1으로 다소 낮은 수치를 보였다. 하지만 치환성 양이온 Na+, Ca2+, Mg2+의 경우에는 우리나라 산림토양과 비교하여 매우 낮은 수치를 보이고 있었으며, 김창환(2012)이 보고한 지리 산 국립공원 내 구상나무림의 토양 조사 결과와 비교하여도 다소 낮은 수준이었다. 따라서 본 조사 대상지인 지리산, 덕유산, 소백산 국립공원 내 구상나무 임분의 건전한 생육을 위해서는 토양 산도의 개선과 치환성 양이온 부족 문제에 대한 개선 대책이 필요할 것으로 판단된다.

Controversy has surrounded the potential impacts of phytoplankton on the tropical climate, since climate models produce diverse behaviors in terms of the equatorial mean state and El Niño-Southern Oscillation (ENSO) amplitude. We explored biophysical impacts on the tropical ocean temperature using an ocean general circulation model coupled to a biogeochemistry model in which chlorophyll can modify solar attenuation and in turn feed back to ocean physics. Compared with a control model run excluding biophysical processes, our model with biogeochemistry showed that subsurface chlorophyll concentrations led to an increase in sea surface temperature (particularly in the western Pacific) via horizontal accumulation of heat contents. In the central Pacific, however, a mild cold anomaly appeared, accompanying the strengthened westward currents. The magnitude and skewness of ENSO were also modulated by biophysical feedbacks resulting from the chlorophyll affecting El Niño and La Niña in an asymmetric way. That is, El Niño conditions were intensified by the higher contribution of the second baroclinic mode to sea surface temperature anomalies, whereas La Niña conditions were slightly weakened by the absorption of shortwave radiation by phytoplankton. In our model experiments, the intensification of El Niño was more dominant than the dampening of La Niña, resulting in the amplification of ENSO and higher skewness.

Ganglioside GD1a is specifically formed by the addition of sialic acid to ganglioside GM1a by ST3 β- galactoside α -2,3-sialyltransferase 2 (ST3GAL2). Above all, GD1a are known to be related with the functional regulation of several growth factor receptors, including activation and dimerization of epidermal growth factor receptor (EGFR) in tumor cells. The activity of EGF and EGFR is known to be a very important factor for meiotic and cytoplasmic maturation during in vitro maturation (IVM) of mammalian oocytes. However, the role of gangliosides GD1a for EGFR-related signaling pathways in porcine oocyte is not yet clearly understood. Here, we investigated that the effect of ST3GAL2 as synthesizing enzyme GD1a for EGFR activation and phosphorylation during meiotic maturation. To investigate the expression of ST3GAL2 according to the EGF treatment (0, 10 and 50 ng/ml), we observed the patterns of ST3GAL2 genes expression by immunofluorescence staining in denuded oocyte (DO) and cumulus cell-oocyte-complex (COC) during IVM process (22 and 44 h), respectively. Expression levels of ST3GAL2 significantly decreased (p<0.01) in an EGF concentration (10 and 50 ng/ml) dependent manner. And fluorescence expression of ST3GAL2 increased (p<0.01) in the matured COCs for 44 h. Under high EGF concentration (50 ng/ml), ST3GAL2 protein levels was decreased (p<0.01), and their shown opposite expression pattern of phosphorylation-EGFR in COCs of 44 h. Phosphorylation of EGFR significantly increased (p<0.01) in matured COCs treated with GD1a for 44 h. In addition, ST3GAL2 protein levels significantly decreased (p<0.01) in GD1a (10 μM) treated COCs without reference to EGF pre-treatment. These results suggest that treatment of exogenous ganglioside GD1a may play an important role such as EGF in EGFR-related activation and phosphorylation in porcine oocyte maturation of in vitro.

The purpose of this study was to review regulatory management of the classification system and scope of veterinary medical devices in Korea. In Korea, the four categories of the classification system for veterinary medical devices (instruments, supplies, artificial insemination apparatus, and others) is somewhat differently than that for human medical devices (instruments, supplies, dental materials, and reagents for in vitro diagnostics). In 2013, veterinary medical devices were classified into approximately 1,400 items, whereas, human medical devices were classified into approximately 2,200 items. Dissimilar to human medical devices, veterinary medical devices have no individual identification codes for effective market management. In conclusion, it is necessary to introduce a device identification code system and re-examine scope of the classification system for veterinary medical devices in Korea.

Prion diseases are a class of transmissible fatal disorders. In order to identify alterations associated with the pathogenesis of prion diseases, several studies have been conducted involving differential gene expression analysis using cDNA libraries, mRNA differential displays, and gene microarrays. These genomic approaches may be useful for identifying genes that are differentially expressed in prion diseases and that may participate directly or indirectly in the pathogenesis of the disease. In this study, we compared the gene expression profiles of normal and CWD-infected TgElk mice using the GeneFishing differentially expressed gene (DEG) screening system and real-time PCR analysis. DEGs were screened using the ACP-based PCR method with GeneFishing synthesis. In order to validate candidate genes, we used quantitative PCR (qPCR), and eleven DEGs were identified. Five of these eleven DEGs were upregulated and two were downregulated in the CWD mice. The DEGs newly identified in this study may be useful for diagnosing and studying the pathogenesis of prion diseases.

The plastic monomer bisphenol A (BPA) is well known as a representative environmental hormones. Recent studies showed that the BPA exposure induced mitochondrial dysfunction and mitochondrial derived reactive oxygen species (mito-ROS). However, changes of antioxidant enzymes expression and ROS production from mitochondria according to the BPA exposure on in vitro maturation (IVM) of porcine oocytes have not been studied. We hypothesized that regulation of ROS production from mitochondria by BPA may play a critical role in meiotic maturation or expansion of cumulus cells in cumulus-oocyte complexes (COCs). To investigate the negative effects of BPA exposure on oocyte maturation, immature pig oocytes were matured in NCSU-23 medium supplemented with BPA (50, 75 and 100 μM) for 44 h. Expectedly, the rates of meiotic maturation and cumulus cell expansion of COCs in the BPA (75 μM) treated group was significantly lower than those of control group (p<0.01). Most of secretion factors expressions from COCs were significantly decreased (p<0.05) in the BPA treated COCs. Next, we investigated the intracellular ROS and mitochondrial specific superoxide production according to the BPA exposure using DCF-DA and mito-SOX staining, respectively. BPA exposure were showed that increasing of both intracellular ROS and mito-ROS, as well as mitochondrial related antioxidant enzymes (sod2, prdx3, prdx5) mRNA expression significantly increased (p<0.01) in COCs. And then, mitochondria membrane potential (MMP) dramatically reduced, and mitochondrial-derived apoptotic factors (bax, bcl-xl, caspase 3) mRNA expressions were increased (p<0.01) in BPA treated COCs. In additon, protein levels of mitochondrial-derived apoptosis genes (AIF, cleaved parp1 and caspase 3) were significantly increased (p<0.05) by BPA exposure. To confirm the reduction of BPA-induced mito-ROS, we used to the mitochondrial-targeted ROS scavenger, mito-TEMPO. Interestingly, addition of mito-TEMPO (0.1 μM) to the BPA pre-treated COCs recovered in meiotic maturation of porcine oocytes. These results demonstrated that BPA exposure was induced increasing of mitochondrial dysfunction, mito-ROS and mitochondrial-mediated apoptosis on pig oocyte maturation. Therefore, we suggest that controlling of mito-ROS plays a critical role in pig oocyte maturation in vitro. These findings will be helpful to solve causes of mitochondrial-related infertility.

In this study, we investigated and analyzed the registration, sales and regulatory management system of in vitro diagnostic veterinary medical reagents (IVDVMRs) in Korea. The registration of IVDVMRs has gradually increased since 2000, and total of 233 products from 58 companies were registered from 1975 to 2014. The market size of IVDVMRs is estimated to be approximately 12 billion Won per year from 2011 to 2013: the export sales and proportion was estimated to be 36.8% as 4.4 billion Won in 2013. Of these products, the ranking of the sales were canine heartworm, bovine tuberculosis, swine fever, porcine reproductive and respiratory syndrome, canine distemper+adenovirus+parvovirus disease, foot and mouth disease, etc. In vitro diagnostic human medical reagents were diverted biological medicine from the medical devices by the revision of the Pharmaceutical Affairs Law Enforcement Regulations in 2014 in Korea. In contrast, in vitro diagnostic devices for animal were still managed as medical devices and biological medicines, respectively. The diagnostic reagents for infectious diseases have neither classification nor grade systems. Good manufacturing practices (GMP) requirements on IVDVMRs were also exempted from the current system. This study suggested that the registration of the IVDVMRs has increased since 2005, and regulations of these devices should be improved for the effective operating system.

Two types of Pt nanoparticle electrocatalysts were composited on Pt nanowires by a combination of an electrospinning method and an impregnation method with NaBH4 as a reducing agent. The structural properties and electrocatalytic activities for methanol electro-oxidation in direct methanol fuel cells were investigated by means of scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and cyclic voltammetry. In particular, SEM, HRTEM, XRD, and XPS results indicate that the metallic Pt nanoparticles with polycrystalline property are uniformly decorated on the electro-spun Pt nanowires. In order to investigate the catalytic activity of the Pt nanoparticles decorated on the electro-spun Pt nanowires, two types of 20 wt% Pt nanoparticles and 40 wt% Pt nanoparticles decorated on the electro-spun Pt nanowires were fabricated. In addition, for comparison, single Pt nanowires were fabricated via an electrospinning method without an impregnation method. As a result, the cyclic voltammetry and chronoamperometry results demonstrate that the electrode containing 40 wt% Pt nanoparticles exhibits the best catalytic activity for methanol electro-oxidation and the highest electrochemical stability among the single Pt nanowires, the 20 wt% Pt nanoparticles decorated with Pt nanowires, and the 40 wt% Pt nanoparticles decorated with Pt nanowires studied for use in direct methanol fuel cells.

In the present study, we investigated the role of binding immunoglobulin protein/glucose-regulated protein, 78-kDa (BIP/GRP78)-regulated endoplasmic reticulum (ER)-stress on meiotic maturation and cumulus cells expansion in porcine cumulus-oocyte complexes (COCs). Previously, it has been demonstrated that unfolded protein response (UPR)- related genes, such as molecules involved in ER-stress defense mechanisms, were expressed in matured oocytes and cumulus cells during in vitro maturation (IVM) of porcine oocytes. However, BIP/GRP78-mediated regulation of ER stress in porcine oocytes has not been reported. Firstly, we observed the effects of knockdown of BIP/GRP78 (an UPR initiation marker) using porcine-specific siRNAs (#909, #693, and #1570) on oocyte maturation. Among all siRNAs, siRNA #693 significantly reduced the protein levels of UPR marker proteins (BIP/GRP78, ATF4, and P90ATF6) in porcine COCs observed by Western blotting and immunofluorescence analysis. We also observed that the reduction of BIP/GRP78 levels by siRNA#693 significantly inhibited the meiotic maturation of oocytes (siRNA #693: 32.5±10.1% vs control: 77.8±5.3%). In addition, we also checked the effect of ER-stress inhibitors, tauroursodeoxycholic acid (TUDCA, 200 μM) and melatonin (0.1 μM), in BIP/ GRP78-knockdown oocytes. TUDCA and melatonin treatment could restore the expression levels of ER-stress marker proteins (BIP/GRP78, p-eIF2α, eIF2α, ATF4, and P90ATF6) in siRNA #693-transfected matured COCs. In conclusion, these results demonstrated that BIP/GRP78-mediated regulation of UPR signaling and ER stress plays an important role in in vitro maturation of porcine oocytes.