정신분열은 대표적인 신경정신성 질환으로 정신분열 환자들은 정신적, 감정적 및 행동적인 장애로 인해 고통받고 있다. 이 연구는 흰쥐를 대상으로 정신분열을 유도함으로써 골격근 조직에서 반응하는 혈관생성 관련 단백질의 변화를 살펴보고 규칙적인 운동의 효과를 알아보고자 수행되었다. 연구의 목적을 위해 흰쥐(SD rats) 총 18마리를 통제(n=6), 정신분열(n=6), 정신분열+운동(n=6) 집단으로 구분하였다. 정신분열 및 정신분열+운동 집단은 총 2주 동안 MK-801처치를 통해 정신분열을 유도하였으며, 정신분열+운동 집단은 4주간 규칙적인 유산소성 운동을 수행하였다. 유산소성 운동은 실험동물용 트레드밀을 이용하여 수행되었으며 HIF-1α 및 VEGF 단백질은 western blot을 통해 관찰하였다. 연구결과, HIF-1α 단백질은 집단간 차이를 보이지 않았다(p>.05). VEGF 단백질의 경우, 통제집단보다 정신 분열 집단에서 유의하게 낮게 나타났으며(p<.01), 정신분열+운동 집단의 VEGF가 정신분열 집단보다 유의하게 높게 나타났다(p<.01). 따라서 골격근 내 혈관생성 관련 단백질들은 정신분열 유도에 의해 낮아질 수 있으며 반대로 운동은 이를 개선하는 효과를 가지는 것으로 사료된다.

붉바리(Epinephelus akaara) 종자생산 시 기형 발생에 의한 손실이 크지만 기형어에 대한 생물 학적 정보는 많지 않다. 본 연구에서는 부화 후 96일 붉바리 치어를 정상 그룹과 두 유형의 기 형 그룹(머리, 턱)으로 나누어 형태형성과 연관된 4개의 주요 유전자(insulin like growth factor 1: IGF-1, bone morphogenic protein 4: BMP4, peroxisome proliferator-activated receptors γ: PPARγ, matrix Gla protein: MGP) 발현을 조사하였다. 각 그룹에서 뇌, 간 및 근육을 잘라낸 다 음 total RNA를 추출한 후 real-time PCR을 사용하여 유전자 발현 차이를 비교하였다(n=20). 부화 후 96일 붉바리 치어에서 IGF-1과 BMP4 유전자는 기형 그룹의 뇌와 간에서 정상 그룹과 비교하여 유의한 발현 차이를 나타냈다(p <0.05). 반면에 PPARγ와 MGP 유전자는 어떤 조직 에서도 정상 그룹과 기형 그룹 사이에 유의한 발현 차이를 보이지 않았다. IGF-1과 BMP4 유전 자는 치어 단계의 붉바리 기형 상태와 관련되어 있는 것으로 보인다.

The baculovirus-insect cell expression system has been widely used method for the recombinant protein expression. The present study has several limitation. In this study, we constructed vectors consisting of transcriptional enhanced factor and promoter that improve the expression level. To confirm the usefulness of these vector system, Human papillomavirus (HPV) VLPs have been expressed by baculovirus hyper expression system. HPV VLPs were purified using a CaptoTM Core 700 (GE Healthcare Life Sciences) chromatography approach. Baculovirus hyper expression system production efficiency was influenced by the HPV VLPs production. HPV VLPs vaccination to BALB/c mice induced the generation of antibody confirmed by ELISA. This study could provide improvements on the vaccine production for the development of VLP vaccines high expression of useful heterologous proteins.

In our previous studies, the cardiac xenotransplantation from an alpha-1,3-galactosyltransferase knockout pig (GT-MCP-MCP) to cynomolgus monkeys showed a mean survival of 38 days. The objective of this study is to genetically upgrade the GT-MCP-MCP pig, to further enhance membrane cofactor protein (MCP) expression and to express an endothelial specific thrombomodulin (TBM). MCP is a complement regulatory protein and TBM is a coagulation inhibitor. As the dicistronic cassette for wild-type-based MCP and TBM concurrent expressions does not show any increase of MCP, we optimized the MCP codon usage (mMCP) and substituted mMCP for MCP. When the mMCP-TBM cassette was transfected to HeLa cells, we were able to find an increased expression of MCP and endothelial cell-specific TBM expression. The cassette was then transfected into ear-skin fibroblasts isolated from one-month-old #23-4 of a GT-MCP-MCP pig, and the cell populations expressing MCP were obtained by MACS cell sorting. We performed a single cell culture of the selected cells, and obtained clones over expressing 90% MCP. The cells of a clone were used as a donor for nuclear transfer and generated GT-MCP/-MCP/mMCP/TBM pig. The transgenic pig was confirmed to be carrying the cells expressing MCP and functioning as an inhibitor against the cytotoxic effect of normal monkey serum, comparable with donor cells. Thus, we believe that the GT-MCP/-MCP/mMCP/TBM transgenic pig would be potential for the prolongation of xenograft survival in the recipients.

The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 μM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

IL-1RAcP는 일명 interleukin-1 receptor accessory protein이라 칭하며 interleukin-1 염증성 사이 토카인과 interleukin-1 receptor I (IL-1RI) 결합체와 복합체를 형성하여 작용한다. IL-1RAcP는 면 역반응, 스트레스 및 세포사멸과 관련이 있다. 본 연구의 목적은 참돔(Pagrus major)을 저수온 (8℃, 33 psu) 및 저염분(20℃, 10 psu) 상태에 노출시킨 후, IL-1RAcP 유전자의 발현을 관찰하는 것이다. 연구결과, IL-1RAcP 유전자의 발현은 저수온(8℃, 33 psu) 및 저염분(20℃, 10 psu) 상태 에서 유의적으로 증가하는 것으로 나타났다. 이 연구결과로서 IL-1RAcP 유전자는 저수온 및 저 염분 등의 환경 스트레스에 대한 생체지표유전자로서 역할을 한다고 제의한다.

The sweetpotato whitefly Bemisia tabaci is one of the most serious pests in the greenhouse. They can infest variouscrop plants even in high warm greenhouse condition in summer. To understand the effect of heat acclimation ability ofB. tabaci, mortality was determined at various temperature conditions of the greenhouse. Adult whiteflies survived evenin 63°C in tomato greenhouse. The median lethal temperature (LT50) of the greenhouse colony was 59.9°C however LT50of the lab colony was 47.6°C. The relative expression level of heat shock proteins (hsp23, hsp70 and hsp90) were significantlylower in the greenhouse colony compare to the lab colony. This result suggests B. tabaci has very high thermal adaptability(acclimation) and its mechanism is associated with heat shock protein system.

We investigate that the impact of freshwater organism exposed to the salinity environment by the frequent rainfall following climate change. To evaluate the stress response following salinity exposure, we assessed the survival rate, molting success rate, the developmental period and mouthpart deformities in Chironomus riparius. In addition, we measured the molecular responses of biomarker gene, gene expression of heat shock protein 70 (HSP70) in C. riparius exposed to salinity after 96 hour. The C. riparius survival rates were showed on time dependent manner and not observed survival organisms above 15 psu at day 4. The pupation and emergence of C. riparius were not seen above 15 psu, and the molting success rate was less than 20% at 10 psu. The developmental retardation of C. riparius was well observed in the pupation and emergence period and was delayed by 4 days at 10 psu compared to the control and 5 psu. The mouthpart deformities after salinity exposure at 96 or 72 hour were observed at 10 psu and 15 psu. The expression of C. riparius HSP70 level was significantly increased exposure to 5 psu and 10 psu. Thus, salinity has been caused to be various ecotoxicological and molecular stress responses on freshwater organisms similar to harmful substances such as EDCs and so on.

Coffee is one of the most familiar beverages to modern human adults, but its bio-physiological effect has not been clearly elucidated. It was known that more than one thousand chemicals were included in the ordinary coffee extract. Among them, the caffein and chlorogenic acid (caffeoylquinic acids) are most abundant and have been investigated by many authors so far. In order to know the real cellular effect of whole coffee extract elements, the dialyzed coffee extract (DCE)1) was made to get coffee elements less than 1000 Da molecular weight, which are freely absorable through gastrointestinal tract. It was directly treated in the culture of RAW 264.7 cells, a murine macrophage lineage. RAW 264.7 cells were treated with DCE equivalent to 2.5 cups of coffee (DCE-2.5), DCE-5, and DCE-10 for 12 hours, and their protein extracts were examined by histological observation and immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the proliferation-related proteins depending on the dose of DCE. DCE-2.5 and DCE-5 enhanced the cellular growth of RAW 264.7 cells by increasing the expression of β-actin, PCNA, Ki-67, MPM2, MAX, cMyc, E2F-1, and Rb-1, and by decreasing the expression of MAD and p21. These proliferation-related proteins were rarely affected by DCE-10. DCE-2.5 and DCE-5 induced the cellular proliferation of RAW 264.7 cells by the signaling of E2F-1 and cMyc, respectively, but these cellular effects almost disappeared in DCE-10. Therefore, it was presumed that the low dose of coffee, DCE-2.5 and DCE-5 might be effective for the proliferation of murine macrophages, RAW264.7 cells, contrast to the high dose of coffee, DCE-10. It was also suggested that the low dose of DCE-2.5 and DCE-5 be helpful to increase the innate immunity in vivo by increasing the cell number of macrophages in contrast to the high dose of DCE-10.

Multidrug resistance (MDR) remains one of the most significant obstacles in various cancer treatment, and this process often involves dysregulation of the number of micro-RNAs. The aim of this study was to explore the role of miR-4708 on the regulation of MDR-1 expression and the regulation of multidrug resistance (MDR) to chemotherapeutic drugs. Luciferase reporter assays demonstrated that miR-4708 directly binds MDR-1 3’-UTR and down-regulated reporter luciferase activity. The mRNA and protein expression levels of MDR1 were significantly decreased following miR-4708 overexpression. Additionally, the accumulation of rhodamine-123 in paclitacel resistant FaDu cells following miR-4708 transfection was significantly increased compared with control, indicating that the efflux capacity was reduced. These results demonstrated that miR-4708 could be involved in the regulation of MDR via targeting MDR-1 and may provide a potential strategy for reversing drug resistance in oral cancer.

The sweetpotato whitefly Bemisia tabaci is a serious pest and virus vector of many crops. To understand thermal tolerance of B. tabaci at molecular level, effects of rapid and acclimated thermal stress were analyzed by measuring mRNA levels of two heat shock proteins (hsp), hsp23 and hsp90, of B. tabaci using quantitative real-time RT-PCR. Mortality of adult whiteflies were 65.5% by 45°C for 1 h but 100% by 50°C for 1 h. However, mortality by subsequent exposures to 35°C for 1 h then 45°C for 1 h was 43.3%. Comparison between rapid and acclimated heat shock treatments showed that different patterns between hsp23 and hsp90 levels. At acclimated condition, hsp90 was higher but hsp23 was lower than rapid heat shock. The results suggest that the heat acclimation response is possibly advantageous to whiteflies that are often exposed to drastic temperature fluctuations.