• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.149-155
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• 2017

Bacterial blight in rice caused by Xanthomonas oryzae pv. oryzae (Xoo) greatly reduces the growth and productivity of this important food crop. Therefore, we sought to increase the resistance of rice to bacterial blight by using a NAC (NAM, ATAF, and CUC) transcription factor, one of the plant-specific transcription factors that is known to be involved in biotic/abiotic stress resistance. By isolating the OsNAC58 gene from rice and analyzing its biological functions related to Xoo resistance, phylogenetic analysis showed that OsNAC58 belongs to group III. To investigate the biological relationship between bacterial leaf blight (BLB) and OsNAC58 in rice, we constructed a vector for overexpression in rice and generated transgenic rice. The expression analysis resulting from use of RT-PCR showed that OsNAC58-overexpressed transgenic rice exhibited higher levels of OsNAC58 expression than wild types. Further, subcellular localization analysis using rice protoplasts showed that the 35S/OsNAC58-SmGFP fusion protein was localized in the nuclei. Thirteen OsNAC58-overexpressed transgenic rice lines, with high expression levels of OsNAC58, showed more resistant to Xoo than did the wild types. Together, these results suggest that the OsNAC58 gene of rice regulates the rice disease resistance mechanism in the nucleus upon invasion of the rice bacterial blight pathogen Xoo.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.169-176
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• 2008

Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.235-243
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• 2002

This study was designed to examine the ability of the bovine (MII) oocytes cytoplasm to support several mitotic cell cycles under the direction of differentiated somatic cell nuclei of bovine, porcine, mouse and human. Bovine GV oocytes were matured in TCM-199 supplemented with 10% FBS. At 20h after IVM, recipient oocytes were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst and their 1st polar body (PB) and MII plate were removed by enucleation micropipette under UV filter. Ear skin samples were obtained by biopsy from an adult bovine, porcine, mouse and human and cultured in 10% FBS added DMEM. Individual fibroblast was anlaysed chromosome number to confirm the specificity of species. Nuclear transferred (NT) units were produced by electrofusion of enucleated bovine oocytes with individual fibroblast. The reconstructed embryos were activated in 5 $\mu$M ionomycin for 5 min followed by 1.9 mM 6-dimethylaminopurine (DMAP) in CR1aa for 3 h. And cleaved NT embryos were cultured in CR1aa medium containing 10% FBS on monolayer of bovine cumulus cell for 8 days. Also NT embryo of 4~8 cell stage was analysed chromosome number to confirm the origin of nuclear transferred somatic cell. The rates of fusion between bovine recipient oocytes and bovine, porcine, mouse and human somatic cells were 70.2%, 70.2%, 72.4% and 63.0%, respectively. Also, their cleavage rates were 60.6%, 63.7%, 54.1% and 62.7%, respectively, there were no differences among them. in vitro development rates into morula and blastocyst were 17.5% and 4.3% in NT embryos from bovine and human fibroblasts, respectively. But NT embryos from porcine and mouse fibroblasts were blocked at 16~32-cell stage. The chromosome number in NT embryos from individual fibroblast was the same as chromosome number of individual species. These results show that bovine MII oocytes cytoplasm has the ability to support several mitotic cell cycles directed by newly introduced nuclear DNA.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.245-252
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• 2002

This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.3
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• pp.391-405
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• 2003

Craniosynostosis, known as a premature fusion of cranial sutures, is a developmental disorder characterized by precocious differentiation and mineralization of osteoblasts in the calvarial sutures. Recent genetic studies have demonstrated that mutation in the homeobox gene Msx2 causes Boston-type human craniosynostosis. Additionally, the phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. Furthermore transcription of osteocalcin, a mature osteoblast marker, is reciprocally regulated by the homeodomain proteins Msx2 and Dlx5. These facts suggest important roles of osteocalcin, Msx2 and Dlx5 genes in the calvarial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we have first analyzed by in situ hybridization the expression of osteocalcin, Msx2 and Dlx5 genes in the developing parietal bone and sagittal suture of mouse calvaria during the embryonic (E15-E18) stage. Osteocalcin mRNA was found in the periosteum of parietal bones from E15, and gradually more highly expressed with aging. Msx2 mRNA was intensely expressed in the sutural mesenchyme, osteogenic fronts and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and the periostem of parietal bones. To further examine the upstream signaling molecules of transcription factor Msx2 and Dlx5, we have done in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of BMP2-, BMP4-soaked beads onto the osteogenic fronts after 48 hours organ culture induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of $TGF{\beta}1$, GDF-6, -7, FGF-2, -4 and Shh did not induce the expression of Msx2 and Dlx5. Taken together. these data indicate that transcription factor Msx2 and Dlx5 play critical roles in the calvarial bone and suture development, and that BMP siganling is involved in the osteogenesis of calvarial bones and the maintenance of cranial sutures through regulating these two transcriotpn factors. Furthermore, different expression patterns between Msx2 and Dlx5 suggest their specific functions in the osteoblast differentiation.

• Journal of the Korean Association of Geographic Information Studies
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• v.17 no.3
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• pp.175-194
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• 2014

This study attempts to identify the spatial distributions of Achnanthes convergens, and elucidate the environmental factors that affect the Periphyton diatom habitat. Data in 250 points of Nakdong river basin are collected between April(primary) and September(secondary) 2012, with the National Institute of Environmental Research's support. We define "clean area" over 10% of Achnanthes convergens appearance, and the others as "non-clean areas". Spatial statistics of Kriging, Hotspot, LISA are used in this study. Results show that 1) 56 points are identified as clean areas in the primary survey, while 41 points are discovered in the following survey; 2) using water quality variables, density of turbidity(clean $101.83{\mu}s/cm$; non-clean $598.48{\mu}s/cm$) and conductivity(clean 1.95 NTU; nonclear 5.58 NTU) are five-fold lower in clean-areas; 3) Habitat and Riparian Factors in Nakdong basin illustrate that natural sand bar, diversity of velocity, sediment condition, levee material, riverside land affect Achnanthes convergens; 4) Hotspots of Achnanthes convergens are located in watersheds, including upper Andong Dam, upper Imha dam, Wi-cheon, Miryang river, Nam river and Hwang river whereas mainstream/downstream of Nakdong river and Keumho river watershed are shown as coldspots.

• Journal of the korean academy of Pediatric Dentistry
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• v.36 no.2
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• pp.245-252
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• 2009

The congenital missing of teeth is common, which takes place since the proliferation and differentiation are not allowed in that tooth bud fail to start development. The purpose of this study is to research incidence rate, number, and missing part of congenital missing teeth, and to study whether a person who has missing teeth has other abnormality of teeth or not. For this study, 1,520 subjects(aged 2.9$\sim$17) who had visited pediatric dentist department of Chonbuk national university dental hospital within 2 years were examined with an panoramic radiograph; exempting third molar missing state. The obtained results are as follows. 1. 8.88% among total subjects show missing teeth; male 9.05%, female 8.64% 2. The most frequently missing permanent teeth were the mandibular second premolars(22.3%). The most frequently missing primary teeth are mandibular lateral incisors(50%). 3. 43.3% patients have one permanent missing tooth, 34.3% have two, and 10.4% have more than six, respectively. In primary teeth, 86.7% patients have one missing tooth, and 13.3% have two missing teeth. 4. 18 patients(13.3%) have missing teeth as well as hyperdontia, while some patients have microdont, ectopic eruption, and fusion teeth.

• KSBB Journal
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• v.23 no.3
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• pp.231-238
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• 2008

The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

• Korean Journal of Biological Psychiatry
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• v.3 no.2
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• pp.222-239
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• 1996

The purpose of this study was to evaluate the effects of alcohol on the psychomotor performance and subjective assessment in healthy Korean adults with acetaldehyde dehydrogenase I(ALDH-I) isozyme variance. A total of 20 male subjects, half with active ALDH-I and the other half with inactive ALDH-I, were selected through both a self-reporting questionnaire examining alcohol sensitivity and the Higuchi's ethanol patch test detecting ALDH-I deficiency. In a doule-blind, placebo-controlled cross-over design, each subject consumed four doses of alcohol(0.25, 0.5, 0.75, 1.0g/kg) and placebo on five separate occasions at weekly intervals, Treatment order was fully balanced using a $5{\times}5$ Latin square, Psychomotor performance tests[coritical flicker fusion threshold(CFF) and choice reaction time(CRT)] and self-estimate questionnaires were conducted at baseline and at time points of 20, 40, 60, 90, 120, 150, 180 minutes after consuming the test drug for 20 minutes, Blood alcohol concentrations(BACs) using breath analyzer were measured at baseline and at time points of 10, 20, 30, 40, 50, 60, 75, 90, 105, 120, 150, 180 minutes after drinking, The BACs and the mean changes in the psychomotor performances and subjective assessments from pre-alcohol baseline, were compared between the two groups. The findings were summarized as follows : 1) BACs were tended to be higher in the inactive group than the active in all of the four alcohol doses. However significant group differences were only after the 0.5g/kg dose of alcohol. 2) The inactive group showed significant impairment in CFFT at most time points alter 0.75 and 1.0g/kg doses of alcohol. 3) In CRT, total reaction time(TRT) significantly prolonged in the inactive group than the active group at 20 minutes after 0.25 and 1.0g/kg doses of alcohol and at 40, 60, 90 minutes alter 0.75g/kg dose of alcohol. In the inactive group, recognition time component significantly increased at 20, 60, 90 minutes after 1.0g/kg dose of alcohol, while movement time component significantly increased at 40, 60 minutes after 0.75g/kg dose of alcohol. 4) Subjective evaluation of the effect of alcohol revealed that physical and mental conditions as well as a self-estimate of the effects of alcohol on performance were significantly worse in the inactive group than the active at some time points alter all of the lour alcohol doses, wihch were more pronounced after 0.75 and 1.0g/kg doses of alcohol. 5) Most of the group differences mentioned above, still remained statistically significant after BAC was entered as a covariate, These findings demonstrated that the alcohol sensitivity is higher in individuals with inactive ALDH-I than those with active ALDH-I both on the subjective assessments and the objective psychomotor performances. Furthermore, these results suggest thai the alcohol sensitivity may be determined by acetaldehyde concentration rather than BAC per se. In future studies, after more accurate genotyping for ALDH-I, the relationships between BAC, acetaldehyde concentration and alcohol sensitivities should be clearly defined.

• Korean Journal of Biological Psychiatry
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• v.3 no.2
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• pp.277-287
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• 1996

Objective : It has been proposed that cognition and related aspects of mental functioning are decreased in depression as well as in alcoholism. The objective of the study was to compare behavioral side effects of paroxetine and amitriptyline in depressed patients accompanied by alcoholism. The focused comparisons were drug effects concerning psychomotor performance, cognitive function, sleep and daytime sleepiness during the first 2 weeks of treatment. Methods : After an alcohol detoxification period(3 weeks) and a washout period(1 week), a total of 20 male inpatients with alcohol use disorder (DSM-IV), who also had a major depressive episode(DSM-IV), were treated double-blind with paroxetine 20mg/day(n=10) or amitriptyline 25mg/day(n=10) for 2 weeks. All patients were required to have a scare of at least 18 respectively on bath the Hamilton Rating Scale far Depression(HAM-D) and Beck Depression Inventory(BDI) at pre-drug baseline. Patients randomized to paroxetine received active medication in the morning and placebo in the evening whereas those randomized to amitriptyline received active medication in the evening and placebo in the morning. All patients performed the various tasks in a test battery at baseline and at days 3, 7 and 14. The test battery included : critical flicker fusion threshold for sensory information processing capacity : choice reaction time for gross psychomotor performance : tracking accuracy and latency of response to peripheral stimulus as a measure of line sensorimotor co-ordination and divided attention : digit symbol substitution as a measure of sustained attention and concentration. To rate perceived sleep and daytime sleepiness, 10cm line Visual analogue scales were employed at baseline and at days 3, 7 and 14. The subjective rating scales were adapted far this study from Leeds sleep Evaluation Questionnaire and Epworth Sleepiness Scale. In addition a comprehensive side effect assessment, using the UKU side effect rating scale, was carried out at baseline and at days 7 and 14. The efficacy of treatment was evaluated using HAM-D, BDI and clinical global impression far severity and improvement at days 7 and 14. Results : The pattern of results indicated thai paroxetine improved performance an mast of the lest variables and also improved sleep with no effect on daytime sleepiness aver the study period. In contrast, amitriptyline produced disruption of performance on same tests and improved sleep with increased daytime sleepiness in particular at day 3. On the UKU side effect rating scale, mare side effects were registered an amitriptyline. The therapeutic efficacy was observed in favor of paroxetine early in day 7. Conclusion : These results demonstrated thai paroxetine in much better than amitriptyline for the treatment of depressed patients accompained by alcoholism at least in terms of behavioral safety and tolerability, furthermore the results may assist in explaining the therapeutic outcome of paroxetine. For example, and earlier onset of antidepressant action of paroxetine may be caused by early improved cognitive function or by contributing to good compliance with treatment.