• BMB Reports
• /
• v.43 no.4
• /
• pp.250-256
• /
• 2010

Natural phosphodiester bond CpG-DNA that contains immunomodulatory CpG motifs (PO-DNA) upregulates the expression of proinflammatory cytokines and induces an Ag-driven Th1 response in a CG sequence-dependent manner in mice. In humans, only phosphorothioate backbone-modified CpG-DNA (PS-DNA) and not PO-DNA has immunomodulatory activity. In this study, we found that liposome-encapsulated PO-DNA upregulated the expression of human $\beta$-defensin-2 (hBD-2) and major histocompatibility class II molecules (HLA-DRA) in a CG sequence-dependent and liposome- dependent manner in human B cells. Of the three different liposomes, DOTAP has the unique ability to enhance the immunomodulatory activity of PO-DNA. In contrast, HLA-DRA and hBD-2 promoter activation can be induced by liposome-encapsulated PS-DNA in a CG sequence-independent manner, depending on the CpG-DNA species. Our observations demonstrate that, when encapsulated with a proper liposome in the immune system, natural PO-DNA has the potential to be a useful therapy for the regulation of the innate immune response.

• Korean Journal of Environmental Agriculture
• /
• v.9 no.1
• /
• pp.1-8
• /
• 1990

The effects of soil environmental conditions on the degradation rates of fenitrothion(O-O-dimethyl O-4-nitro-m-tolyl phosphorothioate) in soils under flooded conditions were examined in the laboratory. Fenitrothion was degraded rapidly and the half life period was within 4 days. Furthermore the degradation was mere rapid under flooded conditions than under upland conditions. The decomposition rate was varied with soils and soil temperatures. Fenitrothion degraded more slowly at 30ppm than at l0ppm. Repeated applications of fenitrothion in soils accelerated the degradation rates. The degradation remarkably increased with amendment of rice straw. However, degradation rates ,were virtually unaffected by the addition of the mixed-fertilizer, the fungicide IBP and the herbicide butachlor. The population of fenitrothion-degrading microbes, which were counted by MPN method, always corresponded with the degradation rates in the soils.

• The Korean Journal of Physiology and Pharmacology
• /
• v.14 no.6
• /
• pp.385-390
• /
• 2010

Excessive extracellular matrix (ECM) accumulation is the main feature of chronic renal disease including diabetic nephropathy. Plasminogen activator inhibitor (PAI)-1 is known to play an important role in renal ECM accumulation in part through suppression of plasmin generation and matrix metalloproteinase (MMP) activation. The present study examined the effect of PAI-1 antisense oligodeoxynucleotide (ODN) on fibronectin upregulation and plasmin/MMP suppression in primary mesangial cells cultured under high glucose (HG) or transforming growth factor (TGF)-${\beta}1$, major mediators of diabetic renal ECM accumulation. Growth arrested and synchronized rat primary mesangial cells were transfected with $1\;{\mu}M$ phosphorothioate-modified antisense or control mis-match ODN for 24 hours with cationic liposome and then stimulated with 30 mM D-glucose or 2 ng/ml TGF-${\beta}1$. PAl-1 or fibronectin protein was measured by Western blot analysis. Plasmin activity was determined using a synthetic fluorometric plasmin substrate and MMP-2 activity analyzed using zymography. HG and TGF-${\beta}1$ significantly increased PAI-1 and fibronectin protein expression as well as decreased plasmin and MMP-2 activity. Transient transfection of mesangial cells with PAI-1 antisense ODN, but not mis-match ODN, effectively reversed basal as well as HG- and TGF-${\beta}1$-induced suppression of plasmin and MMP-2 activity. Both basal and upregulated fibronectin secretion were also inhibited by PAI-1 antisense ODN. These data confirm that PAI-1 plays an important role in ECM accumulation in diabetic mesangium through suppression of protease activity and suggest that PAI-1 antisense ODN would be an effective therapeutic strategy for prevention of renal fibrosis including diabetic nephropathy.

• Environmental Analysis Health and Toxicology
• /
• v.24 no.4
• /
• pp.321-332
• /
• 2009

본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

• Korean journal of applied entomology
• /
• v.19 no.2 s.43
• /
• pp.73-78
• /
• 1980

Some experiment were conducted to evaluate the joint toxic action of carbofuran(2,3-Dihydro-2, 2-dimethyl-benzofuranyl methyl carbamate) and diazinon (0.0-Diethyl-0-(2-isopropyl-6-methyl-e-pyrimidinyl)-0-thionophosphate) against 3rd-instar larvae of the common cutworm, Agrotis fucosa (Butler). Acetone solutions of carbofuran or diazinon or mixture of these two insecticides, were topically applied to the mesonotum of the insect in a constant volume of $1.0{\mu}l$ per larva by means of Burkard microapplicator. Joint toxic action of carbofuran and diazinon for 3rd-instar larva of common cutworm was synergistic. The maximum synergistic effect might be obtained when the ratio of carbofuran to diazinon was 4.8 : 5.2.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.14
• /
• pp.5529-5533
• /
• 2014

Background: The aim of this study was to investigate the effect of a c-myc antisense oligodeoxynucleotide and 5-fluorouracil on the expression of c-myc, invasion and proliferation of HEPG-2 liver cancer cells. Materials and Methods: HEPG-2 cells were treated with lipiosome-mediated c-myc ADSON and 5-fluorouracil. The proliferation inhibition rate and invasion were measured by MTT and invasion assay, respectively. Cell apoptosis was detected by flow cytometry and expression of c-myc by RT-PCR and immunohistochemistry. Results: The proliferation inhibition rate was significantly higher in the antisense oligodeoxynucleotide added-5-fluorouracil group than single antisense oligodeoxynucleotide or 5-fluorouracil group (p<0.05). G0/G1 cells in the antisense oligodeoxynucleotide group and S cells in the 5-fluorouracil groups were significantly increased than that in the control group, respectively (P<0.01). The amplification strips of PCR products in 5-FU, ASODN and combination groups were significantly weaker than that in the control group (P<0.01). The percentage of c-myc-protein-positive cells were significantly lower in antisense oligodeoxynucleotide, 5-fluorouracil and combination groups than that in the control group (P<0.01). Conclusions: A liposome-mediated c-myc antisense oligodeoxynucleotide and 5-fluorouracil can inhibit the proliferation and invasion of liver cancer cells by reducing the expression of c-myc. A c-myc antisense oligodeoxynucleotide can increase the sensitivity of liver cancer cells to 5-fluorouracil and decrease the dosage of the agent necessary for efficacy, providing an experimental basis for the clinical therapy of liver cancer.

• Proceedings of the Korea Society of Environmental Toocicology Conference
• /
• 2002.10a
• /
• pp.156-156
• /
• 2002

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1995.04a
• /
• pp.79-79
• /
• 1995

Although synthetic antisense oligodeoxynucleotides (ODNs) have been used to dissect gene function in vitro, technical difficulties of targeted delivery prevented the use of this approach for investigating the effect of gene products in vivo. Here we report the use of local delivery of antisense transforming growth factor-${\beta}$l (TGF-${\beta}$1) oligonucleotides to decrease the fibrosis in the skin wound. Adult wounds heal with scar-tissue formation, whereas fetal wounds heal without scarring and with a lesser inflammatory and cytokine response. We reasoned that strategy emptying antisense TGF-${\beta}$1 ODNs complementary to TGF-${\beta}$1 mRNA might decrease the scarring in dermal wound of mouse. To evaluate this concept, we tested the effects of antisense ODNs targeted to TGF-${\beta}$1 mRNA by topical application of the chemical on the skin wound. Phosphorothioate antisense ODNs was employed to retard their degradation. When antisense TGF-${\beta}$1 ODNs were applied into the wound site, there was a maked reduction of scar compared with control wound site, These effects of antisense TGF-${\beta}$1 ODNs on the scar formation were associated with decreased expression of TGF-${\beta}$1 gene. However sense TGF-${\beta}$l ODNs had no effect on expression of TGF-${\beta}$1 gene. Also, control wounds healed with excessive fibrosis, whereas the antisense treated wounds healed with less fibrosis. In conclusion, our results indicate that antisense TGF-${\beta}$1 ODNs could be used for amelioating scar formation during wound healing.

• Applied Biological Chemistry
• /
• v.33 no.2
• /
• pp.133-137
• /
• 1990

The effects of soil environmental conditions on the rate of degradation of fungicide IBP (Iprobenfos, S-benzyl O, O-diisopropyl phosphorothioate) in the soils under flooded condintions were examined in the laboratory. IBP in soil was degraded more slowly under flooded conditions than under upland conditions. The degradation greatly varied among soils, and the degradation rate was negatively correlated with the content of soil organic matter. Degradation of IBP was influenced by the soil temperature and the amount of IBP applied. The rate of degradation in soil was remarkably inhibited by the amendment of rice straw but not affected by the treatment of mixed-fertilizer, and insecticide fenitrothion and herbicide butachlor. The degradation of IBP was assumed to be due to microorganisms, especially aerobic microbes, as no degradation was observed in sterilized soil.

• Korean Journal of Environmental Agriculture
• /
• v.1 no.1
• /
• pp.65-70
• /
• 1982

This experiment was conducted to see the effect of repeated application of IBP granular formulation(17%, 0,0-diisopropyl-S-benzyl thiophosphate) on the biodegradation of IBP and diazinon〔0,0-diethyl 0-(2-isopropyl-4-methyl-5-pyrimidinyl) phosphorothioate〕 in silt loam soil with 2.1% organic matter under flooded condition. The persistence of IBP in the soil was shortened by increasing the frequencies of application of the chemical. Enhanced degradation ability in the soil caused by repeated application of IBP was prolonged about 53 days, while the ability did not influence diazinon persistence in the soil. The half-lives of IBP in sterilized soil autoclaved at $121^{\circ}C$ for 30 minutes were about 3 times longer than those in viable soil, suggesting that microbial process was a major factor for IBP degradation in the soil. The total colony number of soil microbes showed little difference between the soils with and without repeated application of IBP. A possible concern of specific soil microorganisms on the pesticide degradation in soil was discussed.