• Fisheries and Aquatic Sciences
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• 제12권2호
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• pp.98-103
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• 2009

The potential utility of the rockbream (Oplegnathus fasciatus) $\beta$-actin 5'-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream $\beta$-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream $\beta$-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the $\beta$-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream $\beta$-actin promoter could be used as a potential regulator for DNA vaccination in this species.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.4907-4911
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• 2016

Glioblastoma multiforme (GBM) is the most aggressive of the gliomas, a collection of tumors arising from glia in the central nervous system. Possible associations between the human cytomegalovirus (HCMV) and the JC virus with GBM are now attracting interest. Our present aim was to investigate the prevalence of the two viruses in Iranian patients from Kerman's cities in the south of Iran. In addition, the expression rates of pp65, large T antigen and p53 proteins were assessed and their relation with GBM evaluated using reverse transcription real time PCR (rReal Time PCR). A total of 199 patients with GBM cancer were enrolled, with $mean{\pm}SD$ ages of $50.0{\pm}19.5$ and $50.7{\pm}19.6$ years for males and females, respectively. The P53 rate was dramatically low suggesting an aetiological role,. Large T antigen expression was found in JC positive samples, while the PP65 antigen was observed in patients positive for CMV and JC. HCMV products and JC virus with oncogenic potential may induce the development of various tumors including glioblastomas. The JC virus produces an early gene product, T-antigen, which has the ability to associate with and functionally inactivate well-studied tumor suppressor proteins including p53 and pRB.

• Reproductive and Developmental Biology
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• 제32권1호
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• pp.15-20
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• 2008

Pronuclear DNA microinjection has been the most universal method in transgenic animal production but its success rate of transgenesis in mammals are extremely low. To address this long-standing problem, we used retrovirus- and lentivirus-based vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of ubiquitously active cytomegalovirus (CMV) promoter to deliver transgenes to bovine embryos. The rate of transgenesis was evaluated by counting EGFP positive blastocysts after injection of concentrated virus stock into the perivitelline space of the bovine oocytes in metaphase II. Among two different types of lentivirus vectors derived from FIV (feline immunodeficiency virus) and HIV (human immunodeficiency virus), the former scored the higher gene transfer efficiency; almost 100% of the blastocysts developed from the oocytes infected with FIV-based vector were EGFP positive. As for the vectors derived Com HIV lentivirus, the transgenesis rate of the blastocysts was reduced to 39%.

• 한국정보통신학회논문지
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• 제18권8호
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• pp.2017-2022
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• 2014

폴리히드론 프로모터, 수포성구내염 바이러스G, 폴리A, 사이토메가바이러스 프로모터, 강화녹색형광단백질, 단백질전달부위 유전자 등을 포함한 새로운 베큘로바이러스 시스템이 제조되었다. 본 재조합벡터 시스템은 인간 섬유아세포에 적용하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구로부터 새롭게 제작된 본 베큘로바이러스 시스템이 유전자의 전이와 유전자 발현 면에서 대조 벡터시스템 보다 고효율을 나타내었다.

• Archives of Pharmacal Research
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• 제23권3호
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• pp.257-260
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• 2000

The tetracycline-controlled transactivator (tTA)-mediated gene activation system was examined in virus infected cells to determine its role in the control of gene expression. In the presence of tTA, the gene expression from the tetO-modified minimal promoter was efficiently activated in the uninfected cells, whereas essentially no activation was observed from the only minimal promoter without the seven direct repeats of 42 bp tetO sequences. However, essentially no activation was observed when only the minimal promoter was used, without the seven direct repetitions of the 42 bp tetO sequences. On the other hand, in the infected cells, a substantial background of $\beta$-glucuronidase expression was detected in the absence of tTA, even though tTA stimulated the gene expression by ~7-fold. This background expression indicates that the sequences within or nearby tetO are involved in the background stimulation of the gene expression by HCMV and HSV-1 . These results suggest that the application of the tTA-mediated gene activation system may not be extremely useful for studying the biological roles of HCMV and HSV genes In the viral replicative cycles, because of the basal activity of the gene expression.

• Journal of Yeungnam Medical Science
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• 제40권3호
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• pp.311-314
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• 2023

Thallium poisoning is usually accidental. We present a case of a 51-year-old woman who was evaluated in June 2018 for myalgia, vertigo, asthenia, and abdominal pain. Physical examination revealed temporal-spatial disorientation, jaundice, and asterixis. The laboratory reported the following: bilirubin, 10.3 mg/dL; aspartate transaminase, 78 U/L; alanine transaminase, 194 U/L; albumin, 2.3 g/dL; prothrombin time, 40%; and platelet count, 60,000/mm³. Serology performed for hepatitis A, B, and C; Epstein-Barr virus; cytomegalovirus; and human immunodeficiency virus was negative, and a collagenogram was negative. Physical reevaluation revealed alopecia on the scalp, armpits, and eyebrows; macules on the face; plantar hyperkeratosis; and ulcers on the lower limbs. Tests for lead, arsenic, copper, and mercury were carried out, which were normal; however, elevated urinary thallium (540 ㎍/g; range, 0.4-10 ㎍/g) was observed. The patient was treated with D-penicillamine 1,000 mg/day and recovered her urinary thallium levels were within normal range at annual follow-up. Thallium poisoning is extremely rare and can be fatal in small doses. An adequate clinical approach can facilitate early diagnosis.

• BMB Reports
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• 제30권2호
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• pp.119-124
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• 1997

Transforming growth $factor-{\beta}\;(TGF-{\beta})$ is a multifunctional polypeptide that exerts biological roles including cell proliferation, differentiation, extracellular matrix deposition and apoptosis in many different cell types. $TGF-{\beta}$, although known as a negative growth regulator, has not been tested in human embryo lung (HEll cells. This study attempts to understand the role of $TGF-{\beta}$ on growth control of HEL cells in relationship to tyrosine phosphorylation pattern of cellular proteins. In density-arrested HEL cells treated with $TGF-{\beta}$, analysis of Western immunoblot showed induction of tyrosine phosphorylation of two major cellular proteins (15 kDa and 45 kDa). In normal proliferating HEL cells with different concentrations of serum, further analysis indicated that the increase in tyrosine phosphorylation of a 45 kDa protein was regulated in serum concentration-dependent manner. However, in proliferating HEL cells treated with $TGF-{\beta}$, tyrosine phosphorylation of 45 kDa was down-regulated. Calcium involvement in the regulation of tyrosine phosphorylation of 45 kDa and 15 kDa proteins was also examined. Tyrosine phosphorylation of 15 kDa protein but not of 45 kDa protein was regulated by exogenous calcium. The level of tyrosine phosphorylation of 15 kDa protein was low at reduced caclium concentration and high at elevated caclium concentration. $TGF-{\beta}$ reversed the pattern of tyrosine phosphorylation of 15 kDa protein. These results suggest that tyrosine phosphorylation of 45 and 15 kDa proteins in HEL cells may be controlled depending on the physiological status of the cells, i.e., low in arrested cells and high in proliferating cells. And the tyrosine phosphorylation of the two proteins appears to be down- or up-regulated by $TGF-{\beta}$.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1705-1712
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• 2016

Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.

• Reproductive and Developmental Biology
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• 제32권3호
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• pp.159-166
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• 2008

본 연구는 vesicular stomatitis virus G glycoprotein (VSV-G)으로 피막이 형성되는 replication-defective MoMLV-based vector를 이용한 hTPO 헝질전환 닭의 생산에 관한 연구이다. 실험에 사용한 retrovirus vector의 구조는 hTPO 유전자의 발현 조절을 위해 internal promoter인 hCMV promoter를 이용하였으며 외래 유전자의 발현을 증가시키기 위해 woodchurk hepatitis virus posttranascriptional regulatory element (WPRE) 서열을 도입하였다. 재조합한 vector는 GP2 293 포장세포에 도입하여 virus를 생산하였으며 이 virus를 이용하여 감염시킨 여러 표적세포에서 hTPO의 발현과 생물학적 활성을 확인하였다. 재조합 hTPO의 생물학적 활성은 시판되고 있는 재조합 hTPO에 비해 우월한 것으로 확인되었다. hTPO 형질전환 닭의 생산을 위하여 1,000배 이상 고농도로 농축된 virus를 stage X 단계의 계란의 배반엽 층에 미세주입하여 대리난각 방법으로 배양하였다. 미세주입한 132개의 계란 중 21일 후에 11개의 계란에서 병아리가 부화하였으며 그중 4마리가 형질전환 개체로 확인되었다. 그러나 생산된 4마리 중 3마리가 부화 후 1개월 이내에 원인불명으로 사망하였다. 본 연구의 의의는 상업적 이용 가능성이 있는 생물학적 활성을 가진 사람의 cytokine 단백질의 대량 생산을 위한 생체 반응기로서의 형질전환 닭 개발의 시례를 제공하는데 있다.

• Clinical and Experimental Pediatrics
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• 제51권9호
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• pp.987-991
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• 2008

목 적 : Kikuchi-Fujimoto disease는 histiocytic necrotizing lymphadenitis라고도 불리는 질환으로 발열과 경부 림프절염을 특징으로 하며 자연 호전되는 경과를 보이는 질환이다. 원인에 대해서는 아직 잘 밝혀지지 않았다. 자연 호전되는 경과 등은 이 질환의 원인으로 바이러스 감염을 추측하게 되고, 고려되는 바이러스로는 대표적으로 EBV, HHV6, HHV8, CMV 등이 있다. 본 연구는 그 중에서 EBV, HHV6를 선택하여 HNL와의 연관성을 밝히고자 하였다. 방 법 : 1999년에서 2005년 사이 부산대학교 병원에서 조직검사에서 KFD으로 진단된 환자 51명을 대상으로 나이, 성별, 발열기간, 침범된 림프절 등에 대하여 의무기록지를 바탕으로 후향적으로 분석하였고 이들의 조직을 이용하여 EBV에 대한 ISH을 시행하고, HHV6에 대한 면역조직화학염색을 각각 시행하였다. 결 과 : 대상환자는 남자가 24명, 여자가 27명이었고 평균 연령은 25.9세였다. EBV에 대한 ISH에서 양성을 나타낸 환자는 51명 중에서 8명으로 15.7%였으며 HHV6에 대한 면역조직화학염색에서 양성을 나타낸 환자는 15명으로 29.4%였다. EBV의 경우 혈청검사(VCA IgG와 VCA IgM)가 같이 시행된 경우가 23명이었고 이중 한 명에서 EBV VCA IgM이 양성이면서 EBV ISH에서도 양성이었다. 결 론 : 본 연구는 HNL의 원인으로서 EBV와 HHV6의 역할을 증명하지 못하였으나, 드물게 HNL의 원인으로 바이러스 감염이 관여할 것으로 추측된다.