This study was to analyse the usability of morphological evaluation of vitrified-thawed oocyte before somatic cell nuclear transfer (SCNT) using Oosight imaging system to show spindle. For the vitrification, in vitro matured bovine MII oocytes were treated by two-step freezing medium without (control group) or with 5 ug/ml cytochalasin-b (CCB group). In Exp. 1, after thawing, recovered oocytes in each treatment group were assessed by live image using Oosight imaging system or/and cytoskeletal protein image using immunostaining. In Exp. 2, in each treatment group the in vitro developmental potential of frozen-thawed bovine oocytes post evaluation using Oosight imaging system and then SCNT was investigated. The SCNT embryos were cultured in CR1aa medium supplemented with 10% FBS, 1 mM EGF and 1 mM IGF at 38.5 C in 5% O2 and 5% CO2 in air for 8 days. In Exp.1, the rates of in vitro survival, morphological good grade and spindle normality of CCB treatment group (91.1%, 54.2% and 55.5%) were better than those of control group (86.1%, 48.5% and 48.5%). After SCNT using vitrified-thawed oocyte, the rates of fusion, reconstructed embryos and blastocyst development were also high in CCB treatment group (66.6%, 36.4% and 3.0%) than control group (60.0%, 27.3% and 0%). These results demonstrated that the identification of morphological spindle image of the vitrified-thawed bovine oocytes using Oosight imaging system helps to predict the SCNT embryo quality.

This study was to investigate pregnancy rate of IVM/IVF/IVC Korean cattle (registered in government) embryos according to transport time course. For the production of embryos, oocytes recovered from slaughtered excellent grade cow and highly motile frozen‐thawed bull semen (purchased from LIMC, KPN#497) was used. In vitro produced embryos were cultured in CR1aa medium for 8 days and some of them were frozen. The rate of average cleavage (>2‐cell) was 83.0% (308/371) and blastocyst rate at day 8 was 34.7% (107/308). Among in vitro produced blastocyst embryos at day 8, most healthy embryos were freshly transferred on production day and some frozen embryos were direct transferred on appropriate day. These embryos were produced in a laboratory, embryo transfer (ET) was planned in 10 areas of the remote island (Jeju) from the laboratory by airplane. Thus, we examined the pregnancy rate in recipient cow according to embryo of transport time course before ET. From embryo transferred 44 recipient cows, overall pregnancy was 40.9% (18/44), these 18 cows were all calved [single, 94% (17/18); twin, 6% (1/18)] and total embryo implantation rate was 26% (19/66). Comparing transport time in the base of 6 hr, pregnancy rate in ET group required less 4 hr (60%, 9/15) was significantly higher than that required more 6 hr (26.3%, 5/19). In direct ET of freezing embryos, the pregnancy rate was 40% (4/10). However, it was difficult to find the meaning of temperature, pH and corpus luteum quality of recipients on comparison of pregnancy rate. When the cell death level of embryos according to storage time in thermos (straw container) before ET was measured by TUNEL staining, apoptotic index was increased with storage time‐dependent. These results demonstrated that long distance transfer of IVM/IVF/IVC embryos is possible and the time of embryo transport is very important for the pregnancy rate on field trial.

본 연구에서는 보건계열 대학생의 열등감을 관리하는 집단상담 프로그램을 활용하여 열등감에 대한 대학생의 긍정적‧부정적 반응에 미치는 효과를 확인하고자 하였다. 2018년 11월 20일에서 2019년 1월 2일까지 S 대학교 보건계열 대학생 총 30명을 대상으로 실험집단과 통제집단으로 나누어, 열등감관리 집단상담 프로그램을 주 2회, 120분씩 총 8회기 실시하였다. 프로그램의 효과성을 검증하기 위하여 열등감반응 척도를 이용하여 사전, 사후, 추후로 실시하였다. 분석 결과, 본 연구는 열등감관리 집단상담 프로그램이 대학생들의 열등감에 대한 부정적 반응을 유의하게 감소시켰으며, 열등감에 대한 긍정적 반응을 유의하게 증가 시킨 것으로 나타났다. 이러한 결과는 열등감관리 상담 프로그램을 통해 또래와의 비교에서 느끼는 대학생들의 열등감에 대한 부정적인 태도를 낮추고 긍정적인 태도를 높일 수 있음을 시사한다.

Percutaneous lumbar epidural adhesiolysis is widely used a treatment for various chronic spinal pain but inadvertent complications of subdural, spinal, or intravascular injection can occur. We report a case of 63-year-old female with unusual pulsatile subdural injection image during attempted lumbar epidural adhesiolysis with fluoroscopy. Pulsatile image confined to the posterior aspect of the spinal canal at L3-4 level was observed. After recognizing subdural injection, we performed epidural adhesiolysis carefully without using steroid and local anesthetics under fluoroscopic guidance. Although unusual, pain physician needed to the understanding of the various subdural fluoroscopic contrast images.

The inflammatory response to infections, such as bacteria and viruses is mediated by multiple host factors. The tumor related-genes are the important cytokines in mammals. However, a number of tumor related-genes are not identified in the rock bream. Here, we have reported the identification and molecular characterization of the tumor related genes. The LPS-induced TNF-α factor 1 and 2 (LITAF1, LITAF2), tumor necrosis factor superfamily member 14 (TNFSF14), tumor necrosis factor receptor superfamily member 14 (TNFRSF14) and translationally controlled tumor protein (TCTP), programmed cell death 10 (PCD10) from rock bream are used for the under investigations. The LITAF1 and LITAF2 consist of 138 and 163 amino acids with a conserved LITAF domain. TNFSF14 and TNFRSF14 comprise 266 and 181 amino acid, respectively. TCTP encompasses of 170 amino acid containing two conserved TCTP signatures. Furthermore PCD10 consists of 210 amino acids. Using quantitative real-time PCR, we have obtained expression analysis results of LITAF1 and 2, TNFSF14, TNFRSF14, TCTP, PCD10 in the various tissue. Compared to the control, the tumor related genes mRNA is detected at a high levels in gill (LITAF1, TCTP), intestine (LITAF2), liver (PCD10), spleen (TNFSF14) and RBC (TNFRSF14). We have also performed gene expression analysis in the kidney, spleen, liver and gill after challenging with Streptococcus iniae, Edwardsiella tarda and Red seabream iridovirus. We have acquired the dynamic regulated mRNA expression to each of pathogen according to the tissue. Expression of tumor related-genes mRNA are significantly increased by infected with pathogens in most of the tissue. But oddly, PCD10 mRNA is expressed significantly decreased by S. iniae infection in all of tissues. Our results reveal that rock bream tumor relatived-genes may be involved in rock bream immune responses to pathogen infections, as well as, they also act like potential biomarkers for innate immunity.

Translationally controlled tumor protein (TCTP) is one of important immune regulator. TCTP has been implicated in cellular processes including the cell growth, cell cycle progression, apoptosis regulation and the protection of cells against various stress condition. In this study, we cloned and characterized TCTP from rock bream (Oplegnathus fasciatus), which is an economically important species in the Korea aquaculture industry. The Full-length of rock bream TCTP (RbTCTP) cDNA was 1041 bp and contained an open reading frame (ORF) of 513 bp, which encoded 170 amino acid sequence. The 5' untranslated region (UTR) was 90 bp while the 3' UTR was 538 bp, containing a polyadenylation signal (ATTAAA). The identity of amino acid sequence was 76%, 75% and 74% in tilapia, orange-spotted grouper and Japanese seaperch, respectively. The positions of microtuble-binding region, Ca⁺ binding region and TCTP signature regions in RbTCTP were similar with those of other fish species and mammalian. The RbTCTP mRNA was expressed highest in the muscle. Expression of TCTP mRNA were significantly variable according to injection of red seabream iridovirus (RSIV), Streptococcosis (S. iniae) and Edwardsiella tarda (E. tarda).

Cereal seeds, sorghum, foxtail millet, hog millet, adlay, and corn are traditionally used as health assistant as well as energy supplying food in Korea. While beneficial phytochemicals to human have revealed in cereals, the information on peptides from cereals is far less accumulated than major reserve protein. Here, we analyzed peptide profiles using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS) in cereal seeds for construction of peptide information and attempted to develop peptide biomarkers for cereal identification. To optimize the analysis condition of SELDI-TOF MS, the effect of dilution factor on binding affinity to protein chips was tested using CM10 and Q10 arrays. Peptide clusters were significantly different at the level of 0.01 p-value. Peak spectra were the most stable in 1:50 of dilution factor in both chip arrays. Numbers of detected peak of 5 cereal seeds were 131 in CM10 and 74 in Q10 array. Each cereal was grouped as a cluster and well discriminated into different cluster in the level of 0.01 p-value. Numbers of potentially identified peptide biomarkers are 11, 13, 9, 5 and 12 in sorghum, foxtail millet, hog millet, adlay and corn, respectively. This study demonstrates that each cereal seed have own distinguishable specific peptides although their function are not identified yet in this study. In addition, the proteomic profiling using SELDI-TOF MS techniques could be a useful and powerful tool to discover peptide biomarker for discrimination and assess crop species, especially under 20 kDa.