• Journal of Life Science
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• v.27 no.5
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• pp.501-508
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• 2017

Bone morphogenetic proteins (BMPs) are multifunctional cytokines that play important roles in a variety of cellular functions. Among BMP family members, BMP2 efficiently promotes osteoblast differentiation through Smad-mediated runt-related transcription factor 2 (Runx2) expression. Several recent studies suggest that BMPs are associated with clock genes, in particular Bmal1. Bmal1 protein heterodimerizes with Clock protein and then induces period 1 (Per1) expression. However, the role of Per1 on osteoblast differentiation remains unclear. In this study, we investigated whether Per1 is involved in osteoblast differentiation. MC3T3-E1 cells were treated with BMP2 for induction of osteoblastic differentiation. Osteogenic maker gene and Per1 mRNA expression were measured using real-time PCR. Interestingly, BMP2 treatment induced Per1 mRNA expression in MC3T3-E1 cells. To further investigate the function of Per1 on osteoblast differentiation, MC3T3-E1 cells were transiently transfected with pCMV-Per1. Per1 overexpression increased Runx2 mRNA and protein levels. Also, mRNA expression and promoter activity of osteocalcin were upregulated by Per1 overexpression. To investigate the effect of interaction between Per1 and osteogenic condition, MC3T3-E1 cells were cultured in osteogenic medium containing ascorbic acid and ${\beta}$-glycerophosphate. Osteogenic medium-induced ALP staining level and mineralization were synergistically increased by overexpression of Per1. Taken together, these results demonstrate that Per1 is a positive regulator of osteoblast differentiation.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.317-324
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans was associated with localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis. The cytolethal distending toxin (CDT) of A. actinomycetemcomitans was considered as a key factor of these diseases is composed of five open reading frames (ORFs). Among of them, An enzymatic subunit of the CDT, CdtB has been known to be internalized into the host cell in order to induce its genotoxic effect. However, CdtB can not be localized in host cytoplasm without the help of a heterodimeric complex consisting of CdtA and CdtC. So, some studies suggested that CdtC functions as a ligand to interact with GM3 ganglioside of host cell surface. The precise role of the CdtC protein in the mechanism of action of the holotoxin is unknown at the present time. The aim of this study was to generate recombinant CdtC proteins expression from A. actinomycetemcomitans, through gene cloning and protein used to investigate the function of Cdt C protein in the bacterial pathogenesis. Materials and Methods: The genomic DNA of A. actinomycetemcomitans Y4 (ATCC29522) was isolated using the genomic DNA extraction kit and used as template to yield cdtC genes by PCR. The amplifed cdtC genes were cloned into T-vector and cloned cdt C gene was then subcloned to pET28a expression vector. The pET28a-cdtC plasmid expressed in BL21 (DE3) Escherichia coli system. Diverse conditons were tested to opitimize the expression and purification of functional CdtC protein in E. coli. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and comfirmed the recombinant CdtC expression by SDS-PAGE and Western Blotting. The expression level of the recombinant CdtC was about 2% of total bacterial proteins. Conclusion: The lab condition of procedure for the purification of functionally active recombinant CdtC protein is established. The active recombinant CdtC protein will serve to examine the role of CdtC proteins in the host recognition and enzyme activity of CDT and investigate the pathological process of A. actinomycetemcomitans in periodontal disease.

• Journal of Aquaculture
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• v.19 no.3
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• pp.166-172
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• 2006

Induced androgenesis, a form of artificial parthenogenesis is an important tool for the generation and use of genetically isogenic or clonal fish strain. An optimized protocol for the genetic inactivation of mud loach (Misgurnus mizolepis) oocytes (i.e. production of androgenetic haploid) was developed using UV-irradiation. Various dose levels of UV significantly affected the fertilizing capacity of the eggs, hatchability of embryos and incidence of haploidy. Based on the extensive examinations of treatment conditions on embryo viability and haploid incidence, the optimum dose level of UV irradiation was turned out to be $10,800ergs/mm^2$ with 56.9% of hatching success and 94.6% of haploidy. The overall yield of putative androgen under optimized treatment condition was more than 50% out of total eggs inseminated. The success of androgenetic reproduction of haploid genome was verified by flow cytometry and PCR amplification of transgene that is exclusive to either one of parental sexes. However, a small portion $(8\sim11%)$ of presumed androgenetic haploid larvae was proven to contain residual DNA fragment(s) from maternal parent.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.390-399
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• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.2
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• pp.113-120
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• 2010

Air-cathode microbial fuel cell consisted of 4 unit cells were operated under batch condition and electricity generation and microbial community structure variation were investigated, depending on separator types and cathode characteristics: A) PEM(Proton Exchange Membrane)-30% Wet proofing Carbon Cloth(WC), B) AEM(Anion Exchange Membrane-WC, C) CEM(Cation Exchange Membrane)-WC, D) PEM-No Wet proofing Carbon Cloth(NC). Maximum power densities of PEM-WC, AEM-WC and CEM-WC were 510.9, 522.1 and 504.8 $mW/m^2$, respectively. But PEM-NC showed relatively lower maximum power density of 218.3 $mW/m^2$. And PEM-WC, AEM-WC and CEM-WC showed similar internal resistances(20.0-28.2 ${\Omega}$). PCRDGGE, PCA and diversity indices showed that uncultured bacteria which reported in previous MFC studies were detected in suspended growth bacteria and attached growth bacteria would be affected not by separator type but by cathode characteristic. Thus, cathode characteristic can be one of the critical factors for power generation in air-cathode MFC using PEM, AEM, and CEM as separator.

• Korean journal of applied entomology
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• v.54 no.4
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• pp.393-400
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• 2015

An insect parasitic mite was found on a larva from Japanese apricot seed. The mite was identified as Pyemotes moseri Yu et Liang (Acarina: Pyemotidae) new to Korea. The host larva was identified as Eurytoma maslovskii using mitochondrial DNA sequencing analysis. We conducted preliminary study on its reproduction and parasitization capacity in laboratory condition. A mated female mite reared on Eurytoma maslovskii larva. We counted and sexed newborn progenies and then eliminated them during periodical observations. To test parasitization capacity, a PCR tube containing mass reared P. moseri and Japanese apricot seeds (assumed bear larva of E. maslovskii) placed in a stainless bath filled with potting soil. One month later, we surveyed the seeds whether the E. maslovskii larva parasitized by mite or not. We repeated this experiment three times with five replications each. Average life span (days from parasitization to the end of reproduction) of gravid females was 24.4 days (n=8). A gravid female reproduced 104.0 female progenies (n=8). Although there were more than seven Japanese apricot seeds per bath containing larva or pupa, we found parasitization only in two seeds.

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.563-575
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• 2005

The aim of present study is to evaluate the influence of adjacent tooth to the microbiology of clinically healthy implant. Control group included patients who had clinically healthy implant and tooth with healthy $periodontium(PD{\leq}3mm)$, test group was composed of patients who had clinically healthy implant and tooth with periodontal pocket(PD>3mm). The criteria of clinically health implant are no pain or discomfort, the restorative suprastructure provide satisfactory fit and function, and the tissue around the fixtures were firm and probing with standard periodontal probe with a rounded tip 0.5mm in diameter resulted in penetration of no more than 5mm when using a force of 0.5N at any location. 38 patients, partially edentulous subjects with endosseous root-form implants were selected. All subjects were medically healthy and had not taken systemic antibiotics and professional plaque control 3 months before sampling. Number of control group is 25(mean age $52{\pm}13$, 26 teeth, 34 implants) and test group is 13(mean age $60{\pm}13$, 13 teeth, 17 implants). All teeth and implants of each patient were examined probing depth(PD), bleeding on probing(BOP), and plaque index(PI), and samples of subgingival plaque were obtained at each site with sterile curet or fine paper points, then the plaque transferred to PBS. Obtained samples were examined for the presence of P. gingivalis, T. forsythensis, and T. denticola by the polymerase chain reaction(PCR). The relationship among clinical parameters and the colonizations by the 3 bacterial species from natural teeth and implants region were analyzed by student t-test. The results of this study were as follows: 1. PD was different in teeth between 2 groups(p<0.05), but the other parameters were not. 2. Statistically significant difference was not found in clinical parameters of implants between 2 groups. 3. All bacterial prevalences of teeth were higher in test group than in control group, and prevalence of T. forsythensis had statistically significant difference between 2 groups(p<0.05). 4. Prevalences of P. gingivalis and T. forsythensis are higher in test group than control group, and that of T. denticola is higher in control group than in test group. But there were no statistically significant differences between 2 groups. In conclusion, there is no statistically significant difference in prevalence of implant microbiology between 2 groups. But if the number of samples increased, it will be possible to find out statistical significance in prevalence of P. gingivalis. It seems that pocket of adjacent tooth influences prevalence of P. gingivalis. These results mean that improvement of the periodontal condition before implantation is very important.

• Journal of Korean Society of Environmental Engineers
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• v.33 no.11
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• pp.847-854
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• 2011

To deal with issues from global climate changes, renewable bioenergy has become important. Algae have been regarded as a good resource for biorefinery and bioenergy, and also have potential capability to remove nutrient and non-decompositional pollutants for wastewater advanced treatment. Although algal-bacterial ecological interaction would be a crucially important factor in using algae for wastewater advanced treatment and resource recovery from wastewater, very little is known about ecological interaction between algae and bacteria in a real wastewater environment. In this study, under a real municipal wastewater condition, we characterized wastewater pollutant treatability and bacterial communities in response to growth of Ankistrodesmus gracilis SAG278-2, which can grow in wastewater and has a high lipid contents. The growth of algal population using the wastewater was inhibited by increase in wastewater bacteria while bacterial survival and cellular decay rate were not influenced by the algal growth. Removals of recalcitrant organic matters and total nitrogen were improved in the presence of algal growth. According to T-RFLP and statistical analysis, algal growth affected time-course changes in bacterial community structures. The following 16S rRNA gene amplicon, cloning results showed that the algal growth changes in bacterial community structure, and that bacterial populations belonging to Sediminibacterium, Sphingobacterium, Mucilaginibacter genera were identified as cooperative with the algal growth in the wastewater.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.180-189
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• 2017

Genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by global agricultural companies. Until now, GM crops have not been cultivated commercially in Korea. Commercialization of GM crops requires a compulsory assessment of environmental risk associated with the release of GM crops. This study was conducted to evaluate the frequency of pollen mediated gene flow from Bt transgenic rice (Agb0101) to japonica non-GM rice (Nakdongbyeo), indica non-GM rice (IR36), and weedy rice (R55). A total of 729,917, 596,318 and 230,635 seeds were collected from Nakdongbyeo, IR36, and R55, respectively, which were planted around Agb0101. Selection of the hybrids was determined by repeated spraying of herbicide and Cry1Ac1 immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using specific primer. The hybrids were found in all non-GM rice and out-crossing ranged from 0.0005% at IR36 to 0.0027% at Nakdongbyeo. All of hybrids were located within 1.2 m distance from the Agb0101 rice plot. The meteorological elements including rainfall and temperature during rice flowering time were found to be important factors to determine rice out-crossing rate. Consideration should be taken for many factors like the meteorological elements of field and physiological condition of crop to set up the safety management guideline to prevention of GM crops gene flow.

• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.2
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• pp.219-225
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• 2019

The purpose of this study was to investigate the odontoblast gene expression related to the subculture speed of supernumerary dental pulp stem cells (sDPSCs). The stem cell is undifferentiated cells which has the ability to differentiate into various cells. Specific stimulation or environment induces cell differentiation, and these differentiation leads to bone or muscle formation. 20 sDPSCs were obtained from 20 children under aseptic condition. During the culture through the 10th passage, the third passage cells which showed short subculture period and 10th passage cells which showed long subculture period were earned. Each cell was divided into differentiated group and non-differentiated group. Quantitative real-time polychain reaction (q-RT-PCR) was performed for each group. The genes related to odontoblast differentiation, Alkaline Phosphatase (ALP), Osteocalcin (OCN), Osteonectin (ONT), Dentin sialophosphoprotein (DSPP) and Dentin matrix acidic phosphoprotein 1 (DMP-1), were measured. Differentiated cells showed more gene expression levels. Undifferentiated cells showed higher gene expression level in 10th passages but differentiated cells showed higher gene expression level in 3rd passages. Cells that showed faster subculture period showed relatively lower gene expression level except for OCN and DSPP.