• BMB Reports
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• v.42 no.4
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• pp.217-222
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• 2009

Controlled gene expression in specific cells is a valuable tool for gene therapy. We attempted to determine whether the lentivirus-mediated Tet-On inducible system could be applied to cancer gene therapy. In order to select the genes that induce cancer cell death, we compared the ability of the known pro-apoptotreic genes, Bax and tBid, and a cell cycle inhibitor, p21cip1/waf1, and determined that Bax was the most effective. For the cancer cell-specific expression of $rtTA2^S$-M2, we tested the matrix metalloproteinase-2 (MMP-2) promoter and determined that it is highly expressed in cancer cell lines, including SNU475 cells. The co-transduction of two lentiviruses that contain sequences for TRE-Bax and $rtTA2^S$-M2, the expression of which is controlled by the MMP-2 promoter, resulted in the specific cell death of SNU475, whereas other cells with low MMP-2 expression did not evidence significant cell death. Our data indicate that the lentivirus-mediated Tet-On system using the cancer-specific promoter is applicable for cancer gene therapy.

• Journal of Ecology and Environment
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• v.44 no.2
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• pp.72-80
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• 2020

Background: Antibiotics are widely used to treat animals from infections. After fertilizing, antibacterials can remain in the soil while adversely affecting the soil microorganisms. The concentration of oxytetracycline (OTC) in the soil and its effect on the soil microbial community was assessed. To assess the impact of OTC on the soil microbial community, it was added to the soil at concentrations of 50, 150, and 300 mg kg^-1 and incubated for 35 days. Results: The concentration of OTC added to the soil decreased from 150 to 7.6 mg kg^-1 during 30 days of incubation, as revealed by LC-MS. The deviations from the control values in the level of substrate-induced respiration on the 5th day of the experiment were, on average, 26, 68, and 90%, with OTC concentrations at 50, 150, and 300 mg kg^-1, respectively. In samples with 150 and 300 mg kg^-1 of OTC, the number of bacteria from the 3rd to 14th day was 2-3 orders of magnitude lower than in the control. The addition of OTC did not affect the fungal counts in samples except on the 7th and 14th days for the 150 and 300 mg kg^-1 contaminated samples. Genes tet(M) and tet(X) were found in samples containing 50, 150, and 300 mg kg^-1 OTC, with no significant differences in the number of copies of tet(M) and tet(X) genes from the OTC concentration. Conclusions: Our results showed that even after a decrease in antibiotic availability, its influence on the soil microbial community remains.

• Journal of the korean academy of Pediatric Dentistry
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• v.38 no.2
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• pp.170-178
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• 2011

The purpose of this study was to assess the prevalence of periodontopathic bacteria and resistance determinants from oral biofilm of children. Subgingival dental plaque was isolated from 87 healthy children, and PCR was performed to determine the presence of 5 periodontal pathogens including P. gingivalis, T. forsythia, T. denticola, F. nucleatum, A. actinomycetemcomitans, and nine resistance genes including tet(Q), tet(M), ermF, aacA-aphD, cfxA, $bla_{SHV}$, $bla_{TEM}$, vanA, mecA. 1. The prevalence of F. nucleatum, T. forsythia. and P. gingivalis was 95.4%, 55.2%, and 40.2%, respectively. In addition. the prevalence of A. actinomycetemc omitans was 5.7%, while T. denticola was 3.4%. 2. In analysis of antibiotic resistance determinants. cfxA, $bla_{TEM}$ and tet(M) were detected in all the samples tested. It was also found that the prevalence of tet(Q) showing tetracycline resistance. $bla_{SHV}$ associated with resistance to ${\beta}$-lactams, ermF exhibiting erythromycin resistance, and, vanA resulting vancomycin resistance was 88.5%, 29.9% 87.4%, and 48.5%, respectively. The aacA-aphD gene showing resistance to aminoglycosides and mecA gene harboring methicillin resistance exhibited the lowest prevalence with 9.2%. 3. In a correlation analysis between periodontopathic pathogens and antibiotic resistance determinants, it was found that there was a significant correlation between T. forsythia and $bla_{SHV}$. Also, P. gingivalis and vanA showed a correlation. Finally, tet(Q) and ermF showed a significant correlation (phi: 0.514) while mecA and vanA also showed a correlation(phi: 0.25).

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.136-139
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• 2009

Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

• Journal of Aquaculture
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• v.21 no.2
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• pp.76-81
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• 2008

We investigated the food-effect for ultra-small rotifer Synchaeta kitina cultured under a individual and community by several diets: 3 single trials(Tetraselmis suecica, TET; Isochrysis galbana, ISO; Marine Chlorella ellipsoidea, CHL) and 3 trials with a mixture of 2 species. The rotifer was cultured on the different feeding concentrations. In the individual cultures, the maximum number of offsprings and maximum lifespan of the female investigated to 5.8 inds. and 12.7 days in TET trial, respectively. Values of the developmental phases of the rotifer fed with T. suecica were higher than those of trials without T. suecica. Also it approached faster to maturation level. In the community cultures, the maximum density of TET+CHL trial elevated up to 1,569 inds./mL. But, CHL and ISO showed a poor growth rate and maximum density. The offsprings of the female increased continuously when fed by T. suecica trial, up to $10{\times}10^3$ cells/ind./day. As the quantity of supplied diet was lowered their lifespan were decreased. But, the maximum density and growth rate in the community cultures showed the highest value in the $10{\times}10^3$ cells/ind./day. The efficient food for mass culture of S. kitina was T. suecica, and optimum concentration of their food was 10,000 cells for an individual.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.50-50
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• 2003

모유에 존재하는 유지방구의 막을 구성하는 주된 당단백질인 하나인 lactadherin(과거에는 BA46로 일컬어짐)은 rotavirus에 의한 감염증상을 예방하는 것으로 보고되고 있다. 본 연구에서는 retrovirus vector system을 이용하여 Chinese Hamster Ovary (CHO) 세포에 tetracycline에 의해 발현이 제어되는 promoter 하의 lactadherin 유전자를 전이시킨 후lactadherin이 tetracycline에 의해 발현이 유도되는지의 여부를 실험하였다. 먼저 기초 실험으로 E. coli LacZ유전자를 이용하여 tetracycline에 의한 유도 여부를 조사하였다. Tet-On과 RevTRE-LacZ retrovirus를 co-infection시킨 NIH3T3 세포는 doxycycline (tetracycline 유도체)의 투여량에 비례하여 E. coli LacZ 유전자의 발현 정도가 증가하는 양상을 나타내었는데, 최대의 발현에 대한 doxycycline 농도는 1 $\mu\textrm{g}$/$m\ell$ 이상으로 나타났다. 이 예비실험의 결과를 바탕으로 Tet-On과 RevTRE-Ltd retrovirus vector를 이용하여 사람의 lactadherin 유전자의 유도적 발현을 검정하였는데, CHO 세포에서 lactadherin 유전자의 유도적 발현을 RT-PCR 기법을 이용하여 확인하였다. 표적세포 내에서 외부에서 도입된 유전자가 지속적으로 발현될 경우 심각한 생리적 부작용을 야기시킨다는 사실을 감안할 때, 본 실험의 결과는 유전자 치료와 형질전환동물의 생산에 크게 도움이 될 것으로 예상된다.

• Journal of the Korean Magnetics Society
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• v.19 no.3
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• pp.106-112
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• 2009

$Ni_{1-x}Mg_xFe_2O_4$ ferrite system was studied by using X-ray diffraction and $M{\ddot{o}}ssbauer$ spectroscopy. The samples were prepared by ceramic sintering method with Mg content x. The X-ray diffraction patterns of samples show phase of cubic spinel structure. There are no remarkable changes of lattice constants in $Ni_{1-x}Mg_xFe_2O_4$ ferrite system. The $M{\ddot{o}}ssbauer$ spectra were consisted of two sets of six lines, respectively, corresponding to $Fe^{3+}$ at tetrahedral and octahedral sites. The magnetic hyperfine field of samples was decreased as increasing Mg contents x in both sites and it was shown Yafet-Kittel magnetic structure. $NiFe_2O_4$ was shown complete inverse spinel, but $NiFe_2O_4$ was shown partial inverse spinel which absorption area ratio (oct/tet) was 1.449 in $M{\ddot{o}}ssbauer$ spectrum.

• Physical Therapy Rehabilitation Science
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• v.9 no.2
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• pp.105-112
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• 2020

Objective: To investigate the effects of training using a trunk control robot (TCR) system combined with conventional therapy (CT) on balance and gait abilities in persons with chronic stroke. Design: Two-group pretest-posttest design. Methods: Thirty-five subjects with chronic stroke were randomly assigned to either the TCR group (n=17) or the trunk extension-training (TET) group (n=18). Both groups performed CT for 30 minutes, after which the TCR group performed TCR training and the TET group performed trunk extension training for 20 minutes. Both groups performed the therapeutic interventions 3 days per week for 6 weeks. Balance ability was evaluated using the Berg Balance Scale (BBS), and the Timed Up-and-Go (TUG) test. Gait ability was measured using the 10 m Walk Test (10MWT) and the NeuroCom Smart Balance Master. Results: TCR group showed significant improvements in static balance (weight bearing) and dynamic balance (weight shifting speed, weight shifting direction, BBS, and TUG), 10MWT, gait speed, and step width (p<0.05); step length was not significant. The TET group showed a significant partial improvement of dynamic balance (weight shifting speed, weight shifting direction, BBS, and 10MWT (p<0.05), but the improvements in static balance, TUG, gait speed, and step width and step length was not significant. Additionally, significant differences in static balance, dynamic balance (weight shifting speed, weight shifting direction, BBS, and TUG), 10MWT, gait speed, and step width were detected between groups (p<0.05). Conclusions: TCR training combined with CT is effective in improving static and dynamic balance, as well as gait abilities in persons with chronic stroke.

• Korean Journal of Poultry Science
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• v.36 no.2
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• pp.177-186
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• 2009

It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

• Journal of the Mineralogical Society of Korea
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• v.20 no.4
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• pp.367-376
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• 2007

Aenigmatite, $Na_4(Fe^{2+},Ti,Fe^{3+}){_{12}}(Fe^{3+},Si){_{12}}O_{40}$, is a common constituent of sodium-rich alkaline igneous rocks and is classified a an open-branched single-chain silicate. $M\ddot{o}ssbauer$ spectroscopy of three natural aenigmatite specimens were done and the detailed crystal chemistry was obtained. Fitting of $M\ddot{o}ssbauer$ spectra led to the resolution of nine peaks. They consist of three doublets of $Fe^{2+}/oct$ and one merged peak at low velocity matching to two small peaks at high velocity which were assigned to $Fe^{3+}/tet\;and\;Fe^{2+}/oct$, respectively. Using the peak area for $Fe^{2+}\;and\;Fe^{3+}$ peaks, analytical data were recalculated. Precise assignment of $Fe^{2+}\;and\;Fe^{3+}$ ions in tetrahderal and octahedral sites revealed detailed crystal chemistry of aenigmatite. The existence of significant amounts of $Fe^{3+}/tet$ indicates that $Fe^{3+}$ has preference over $Al^{3+}$ for the tetrahedral sites. Crystal chemistry of aenigmatite (AEN1) yields the formula of $(Na_{3.97}Ca_{0.03})(Ca_{0.11}Mn_{0.59}Fe^{2+}{_{8.07}}Ti_{2.07}Mg_{0.70}Fe^{3+}{_{0.43}}Al_{0.04})(Fe^{3+}{_{0.56}}Al_{0.18}Si_{11.26})O_{40}$.