본 연구에서는 azoxymethane (AOM)과 dextran sodium sulfate (DSS)로 유도된 대장 발암과정에 대한 셀레늄의 방어 효과를 조사하였다. 셀레늄 결핍(0.02 ppm Se), 정상(0.1 ppm Se), 과다(0.5 ppm Se)사료를 12주간 식이로 급여하여 혈액검사와 대장암 발생의 초기단계인 aberrant crypt foci (ACF)수를 측정했으며, 암 발생율을 조사하였다. ICP-AES 를 사용하여 간의 셀레늄 농도를 측정하였으며, 또한 셀레늄포함 항산화효소인 glutathione peroxidase (GPx) 활성을 알아보았다. 또한 TUNEL assay와 PCNA, β-catenin에 대한 면역조직 염색을 수행하였다. ACF 수 및 종양 발생률에 있어서, 셀레늄과다사료를 급여한 군이 정상셀레늄사료를 급여한 군보다 낮았으며, 셀레늄결핍사료를 급여한 군은 오히려 ACF 수 및 종양 발생률이 높았다. GPx 활성은 셀레늄의 섭취가 과다한 군에서 높게 나타났으며, 이 때, TUNEL 에서 apoptotic positive cell이 증가하는 것을 확인했다. 또 한 셀레늄의 섭취가 과다한 군에서 PCNA와 β-catenin의 발현이 감소됨을 볼 수 있었다. 본 마우스 모델실험에서 셀레늄은 여러 기전에 의해 대장암 발생을 억제할 수 있을 것으로 사료된다.

The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP4, GP5, and M. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the GP4, GP5, and M with SUMO (Small ubiquitin-related modifier), and expressed the fused gene in Bm5 cells and silkworm larvae. Expression of the proteins were analyzed by 12% SDS-PAGE and western blotting using 6xHis tag and porcine anti-PRRSV antibodies. In results, SUMO fused proteins were expressed at a high level in Bm5 cells. The levels of protein using the silkworm larvae is higher than that using Bm5 cells. The fused protein was purified by Ni-NTA affinity chromatography. This study demonstrated that SUMO, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

Apolipophorin-III (apoLp-III) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insects. Recently, apolipophorin-III in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of the apoLp-III in insects. To investigate the genes which have a relationship with apoLp-III in fall webworm larvae, we reduced endogenous Hc apoLp-III mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-III reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-III, which induced an elevated level of superoxide anion in Hyphantria cunea larvae. The observed effect of Hc apoLp-III RNAi suggests that Hc apoLp-III is related to the action/expression of antioxidants, especially MnSOD.

A new insect member of the STAT family of transcription factors (HcSTAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and transcribed in hemocyte, fat body, midgut, epidermis, and Malpighian tubule. Especially, hemocyte and Malpighian tubule showed transcriptional activation of HcSTAT upon Gram-negative and -positive bacteria challenge. Gram-negative and -positive bacteria challenge specifically results in nuclear translocation of HcSTAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vivo treatment with sodium orthovanadatetranslocates HcSTAT to the nucleus in hemocyte cells.

The Classical Swine Fever Virus (CSFV) is a member of the Pestivirus genus of the Flaviviridae. The genome of CSFV is a positive single-stranded RNA molecule 12.3 kb and contains a single large open reading frame (ORF). The polyprotein composed of eight nonstructural and four structural proteins (nucleocapsid protein C and three envelope glycoprotein E0, E1 and E2). E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. To determine the characteristics of the CSFV, LOM strain, we investigated the nucleotide sequence of the glycoprotein E0, E1 and E2. Comparison of the LOM with the other strains revealed nucleotide sequence identity ranging from 97 to 98%. Expression of the glycoprotein E2 was identified by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies in Sf21 cells. The expression levels of glycoprotein E2 were observed from day 3 and 5 days maximum. In addition, its expression efficiency by media and cell line was investigated. The result showed that High-Five cells and Grace’s insect media for Sf21 were the best conditions for the expression of the glycoprotein E2.

Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

Embryonic stem (ES) cells are known to have an infinite proliferation and pluripotency that are associated with complex processes. The objective of this study was to examine expression of genes differentially regulated during differentiation of human ES cells by suppression subtractive hybridization (SSH). Human ES cells were induced to differentiate into neural precursor cells via embryoid body. Neural precursor cells were isolated physically based on morphological criteria. Immunocytochemical analysis showed expression of pax6 in neural precursor cells, confirming that the isolated cells were neural precursor cells. Undifferentiated human ES cells and neural precursor cells were subject to the SSH. TPX2 (Targeting Protein for Xklp2 (Xenopus centrosomal kinesin-like protein 2)) was identified, cloned and analyzed during differentiation of human ES cells into neural lineages. Expression of TPX2 was gradually down-regulated in embryoid bodies and neural precursor cells relative to undifferentiated ES cells. Targeting Protein for Xklp2 has been shown to be involved in cell division by interaction with microtubule development in cancer cells. Taken together, result of this study suggests that TPX2 may be involved in proliferation and differentiation of human ES cells.

Somatic cells nuclear transfer (SCNT) is a useful tool in studies of developmental biology and animal cloning. However, SCNT experiments only are allowed to skilled technical experts. In this experiment, laser-assisted zona pellucida piercing tool (LASER) was applied in murine SCNT. LASER minimized the use of piezo-driven micromanipulator (PIEZO), reducing chances of problems caused by PIEZO pulses. LASER reduced time that took to pierce zona pellucida in removal of nucleus from oocyte and somatic cell injection, which might have taken longer time with PIEZO. Time and difficulties that took researcher of equivalent skilled for their experiments were decreased with LASER, and this might affect the improvement of embryonic development. (LASER, 6.2% versus PIEZO, 2.9%; P<0.05). Thus, these data support that the use of LASER can be used for zona pellucida piercing in murine SCNT program as an alternative to PIEZO.

The present study investigated the effects of follicle stimulating hormone (FSH) and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. Oocytes were recovered from mongrel female ovaries in various reproductive states; follicular, luteal or anestrous stage. Oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.5, 1.0 or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0 or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 hr to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with 1.9 μM. Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. More (p<0.05) oocytes matured to MII stage when follicular stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the control, 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (p<0.05) maturation rate to MII stage was observed in follicular stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG supplemented groups (0 to 1.5%). However, the combination of FSH and hCG did not improve the nuclear maturation rate of canine oocyte (2.4 %) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to MII stage.

근접방사선치료는 일반적으로 외부방사선치료와 병행하여 수행되고 치료단계가 매우 복잡하며 이로 인 해 방사선 사고가 발생될 수 있다. 본 연구에서는 이를 해결하기 위해 근접방사선치료에 사고유형과 영향 분석(Failure mode and effects analysis, FMEA) 방법을 적용하여 프로세스 맵을 구성하고 이를 기반으로 각 치료단계에 대한 위해도를 산출하였다. 프로세스 맵은 “외래 및 진료”와 “근접방사선 모의치료”, “CT 모의 치료”, “근접방사선치료계획”, “방사선치료”로 총 5단계로 구성하였으며, 각 치료단계를 세분화하여 세부단 계를 작성하였다. 위해도를 산출하기 위해 의사와 의학물리사, 선량설계사, 방사선사, 간호사가 참여하여 세부단계마다 발생빈도와 심각도, 불검출도를 평가하였다. 전반적으로 프로세스 맵은 각 치료단계마다 환 자 신원 확인 절차가 우선적으로 수행되며, 이는 다른 환자로 오인하여 서로 다른 치료계획이 수립되어 방 사선사고가 발생될 우려가 있다. 프로세스 맵을 기반으로 작성한 세부단계에 대해 위해도를 평가한 결과, 전반적으로 “외래 및 진료”와 “근접방사선치료계획” 과정이 높은 위해도로 평가되었다. 직종마다 평가한 위해도는 서로 다른 경향을 보였으며, 간호사는 방사선치료를 제외한 모든 과정이 55점 이상의 위해도를 보였으며, “근접방사선 모의치료” 과정이 88.8점으로 가장 높았다. 방사선치료를 수행하는 의료기관마다 치 료단계가 다소 차이가 있으므로 해당 기관에 대한 프로세스 맵을 작성하고 위해도를 산출하여 중점관리 항목을 집중적으로 리스크 관리가 수행되어야 할 것으로 생각된다.

This study aimed to analyze difference in clinical findings, including coronary artery complications, in patients with Kawasaki disease and respiratory symptoms with several respiratory infections. We studied 182 pediatric patients diagnosed with Kawasaki disease. Examinations for respiratory viral polymerase chain reaction were conducted in the group of patients with respiratory symptoms. Echocardiography was perfomed by a pediatric cardiologist, and laboratory findings were evaluated. Clinical manifestations and laboratory findings based on medical records were compared. There were no differences between patients with and without respiratory viral infections with respect to age, male-female ratio, coronary artery complications, Kawasaki disease-specific clinical manifestations, duration of fever, duration of hospitalization, or recurrence rate. There was a significant difference in C-reactive protein levels (55.6 vs. 73.9 mg/L) between the two groups, but the other laboratory findings. The rate of respiratory infections in pediatric patients with Kawasaki disease was similar to those reported in previous studies, and clinical manifestations and laboratory findings were not significantly different between the groups.

Background : Although ginseng has various bioactive compounds in it, there is lack of study on the variations of bioactive compounds in ginseng according to the cultivation soil and the applied fertilizer types (or amount). Therefore, this study aims to examine the variations of 37 fatty acids (FA) and 8 vitamin E (Vit-E) vitamers in 6-year-old ginseng root cultivated in different soil types with different fertilizers regimes. Methods and Results : The profiling of 37 FAs and 8 Vit-E vitamers in 6-year-old ginseng roots was measured by gas chromatography coupled with a flame ionization detector, and then these results were statistically analyzed with chemometrics. The FA and Vit-E content in ginseng roots varied significantly with respect to soil cultivation conditions due to organic fertilizer types and amounts used. Unsaturated FA in ginseng is approximately 2.7 fold higher than the saturated FA. Linoleic, palmitic, and oleic acids were the most abundant FAs found in the ginseng roots. Also, the major Vit-E vitamer found in ginseng root is α-tocopherol. In particular, the application of rice straw compost or food waste fertilizer was increased to create nutritionally desirable FAs and bioactive Vit-E in ginseng root. In addition, phytonutrient profiling coupled with chemometrics can be used to discriminate the cultivation conditions of ginseng. Conclusion : This study extends our understanding about the variations of FA and Vit-E in ginseng root depending on cultivation conditions. Hence, these results can be useful as basic information for reliable ginseng production containing high amounts of phytonutrients in a paddy-converted field.

Background : Corrosion is one of the most devastating problems faced by most industries. Mild steel has played a vital role in various fields due to the excellent mechanical properties of mild steel such as low density, high strength-to-weight ratios, good environmental stability, high thermal conductivity, and corrosion resistance. Methods and Results : The total phenolic contents (TPC) and total flavonoid contents (TFC) of the methanolic extract of C. grandiflora and R. verniciflua leaf have been examined, and its corrosion inhibition performance was investigated by weight loss and electrochemical impedance spectroscopy (EIS) and polarization measurements. The surface morphology of mild steel was analyzed by scanning electron microscopy (SEM) equipped with energy dispersive X-ray spectroscopy (EDX) and by atomic force microscopy (AFM). The percentage composition of polyphenolic compounds was found to be higher in C. grandiflora and R. verniciflua plant extracts, and it was proved to be a superior, eco-friendly, and anti-corrosive inhibitor for mild steel in 1M of H2SO4. The Tafel polarization studies indicate that the plant extract is a mixed-type inhibitor. Scanning electron microscopy/energy -dispersive X-ray spectroscopy (SEM-EDS), and atomic force microscopy (AFM) studies confirmed the formation of a protective film on the metal surface. The corrosion inhibition of the C. grandiflora and R. verniciflua plant extracts was characterized by Fourier transform infrared (FT-IR), UV-visible spectra, and wide-angle X-ray diffraction (XRD) studies; these show the strong interaction between the metal surface and the inhibitor. Conclusion : The methanolic extract was prepared the two different plants like C. grandiflora, and R. verniciflua was studied the corrosion inhibition on the mild steel specimen in acidic medium through various methods involving weight loss measurements, EIS, and potentiodynamic polarization. The results shows that the C. grandiflora, and R. verniciflua plant extracts illustrate an effective corrosion inhibitor for mild steel with good anticorrosion properties in acidic environmen