Molecular Identification and Real-time Quantitative PCR (qPCR) for Rapid Detection of Thelohanellus kitauei, a Myxozoan Parasite Causing Intestinal Giant Cystic Disease in the Israel Carp

Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

Cytotoxicity of OKT9 ScFv-Diphtheria Toxin Fusion Immunotoxin on Human Brain Tumor Cell Lines

OBJECTIVE: Immunotoxin therapy is a novel approach for the treatment of tumor, and it has been successfully used in the central nervous system. The purpose of this study is to evaluate the cytotoxicity of OKT9 ScFv-Diphtheria toxin fusion immunotoxin on various human brain tumor cell lines. METHODS: Immunotoxin which was composed of OKT9 ScFv and Diphtheria toxin was made. Its cytotoxicity on glioblastoma cell lines(U87MG, U118MG) and medulloblastoma cell line(TE671) was tested and compared with anti-cancer chemotherapeutic agents. And we also examined the relationship between its cytotoxicity and transferrin receptor expression. RESULTS: It showed most cytotoxicity on U87MG cell line and nearly no effect on U118MG cell line, moderate cytotoxicity on TE671 cell line in sixteen hours exposure experiment. In continuous exposure experiment, it showed moderate cytotoxicity on U118MG cell line, but showed strong cytotoxicity on other cell lines comparable or higher than anti-cancer chemotherapeutic agents. The relationship between its cytotoxicity and transferrin receptor expression was tested using flow cytometry, but no direct relationship could be found. CONCLUSION: Collectively, the result shows the cytotoxic effects of OKT9 ScFv-Diphtheria toxin fusion immunotoxin against various human brain tumor cell lines in continuous exposure experiment. Therefore, we suggest that this immunotoxin could be developed as a potential immunotherapeutic agent in the treatment of various human brain tumor clinically.

Virtual Laparoscopic Cholecystectomy Simulator Using Force-Feedback Device / 대한의료정보학회지

For the minimally invasive surgery, endoscopic surgery has been being a useful technique in diagnosis and treatment of many kind of human diseases and injuries. But, performing endoscopic operations demand expert skills which allow surgeon to learn navigating complex organs in the body through limited view of endoscope as well as manipulating surgical instruments. So the virtual reality based endoscopic surgery simulator has been great increasingly developed with improvements in computing power, graphic hardware, and haptic device in the recent few years. A good balance between surgical realism and real-time interactive rates of simulation is one of most challenging problems in surgical simulation. And when coupled with precise computations of the forces and deformation of organs, it may be possible for the surgeon to feel haptic sensations close to reality. We describe the virtual laparoscopic cholecystectomy simulator using force-feedback device. A goal of this paper is to improve the realism of simulator through real-time interaction between force-feedback and visual-feedback. To calculate deformation of virtual objects separately from the force-feedback, because the latter must be updated much more frequently, the two feedbacks are implemented in each processor. And pre-calculated data set which used for reducing amount of calculation about deformation results in improvement of the real-time interactive rates between the two feedbacks. When virtual surgical tool, manipulated by the user, was collided with isosurface based organ, the surface was deformed in real-time and the graphics, including tissue deformation, performed at 20~30 frames per second. Simultaneously, force-feedback was transmitted to the user at about 500 times per second, therefore we could implement real-time interactive virtual laparoscopic surgery simulator.