• Journal of Korean Neurosurgical Society
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• v.29 no.4
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• pp.471-476
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• 2000

Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

• Korean Journal of Microbiology
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• v.53 no.4
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• pp.286-291
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• 2017

In eukaryotic cells, histone modification is an important mechanism to regulate the chromatin structure. The methylation of the fourth lysine on histone H3 (H3K4) by Set1 complex is one of the various well-known histone modifications. Set1 complex has seven subunits including Swd2, which is known to be important for H2B ubiquitination dependent on H3K4 methylation. Swd2 was reported to regulate Set1's methyltransferase activity by binding to near RNA recognition motif (RRM) domain of Set1 and to act as a component of CPF (Cleavage and Polyadenylation Factors) complex involved in RNA 3' end processing. According to the recent reports, two functions of Swd2 work independently of each other and the lethality of Swd2 knockout strain was known to be caused by its function as a component of CPF complex. In this study, we found that Swd2 could influence the Set1's stability as well as histone methyltransferase activity through the association with RRM domain of Set1. Also, we found that ${\Delta}swd2$ mutant bearing truncated-Set1, which cannot interact with Swd2, lost its lethality and grew normally. These results suggest that the dual functions of Swd2 in H3K4 methylation and RNA 3' end processing are not independent in Saccharomyces cerevisiae.

• Journal of Radiation Industry
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• v.10 no.4
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• pp.211-217
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• 2016

We examined damages from gamma-irradiaion and determined the optimal gamma-ray dose for mutation breeding in lentil (Lens culinaris L.) bean. Four individual lines (L-C, L-2, L-8 and L-9), that have remarkable adaptability in South Korea were gamma-irradiated at doses of 50, 70, 100, 200, 300, 400, and 500 Gy. The germination rate of seed decreased as the dose increased over 50 Gy in all lines. However, $LD_{50}$ and $RD_{50}$ were different among lines. The median lethal doses($LD_{50}$) were approximately 127 (L-C), 74 (L-2), 95 (L-8), and 144 (L-9) Gy. The median reduction doses($RD_{50}$) for plant height, number of leaves, root length, and flash weight were 156, 176, 150, and 180 Gy for L-C, 253, 198, 127, and 142 Gy for L-2, 188, 175, 200, and 190 Gy for L-8, and 162, 210, 224, and 184 for L-9, respectively. The growth characteristics of the $M_1$ generation decreased as the dose increased over 70 Gy. The optimal doses of gamma irradiation for mutation breeding of lentil were determined to be 70 Gy (L-2, L-8) and 100 Gy (L-C, L-9). We performed the comet assay to observe nuclear DNA damage induced by gamma-irradiation. In comet assay, a clear difference was identified over 100 Gy treatments. With increasing doses of gamma-ray in the range of 50 to 500 Gy, the rate of head DNA was decreased significantly from 97.5% to 81.6%. Tail length was consecutively increased from $1.9{\mu}m$ to $17.4{\mu}m$. Our result provides basic information for construction of mutant pools in lentils.

• Journal of Radiation Industry
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• v.7 no.2_3
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• pp.101-108
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• 2013

To develop new varieties of rapeseed (Brassica napus L.), the seeds of three varieties, 'Naehan', 'Tammi', and 'Halla' were irradiated with proton ion beams and gamma rays with 0 to 2,000 Gy. We had selected 9 lines in $M_5$ generation, and their useful characteristics were investigated by progressing from $M_6$ to $M_7$ generation for checking uniformity and stability. The 9 lines selected in $M_5$ generation were maintained their characteristics in terms of flowering date, maturing date, and plant height through $M_6$ to $M_7$ generations. Especially, 2 lines of NP600-1-1-198-2-1 and NP1000-13-2-362-4-1 selected in $M_5$ generation derived from 'Naehan' had characteristics of early maturity and shorter stem than original variety, and they also were maintained characteristic of early maturity such as 10~11 days earlier flowering date and 6~9 days earlier maturing date through $M_6$ to $M_7$ generations. For stem length, they showed characteristics of shorter stem in 2 lines of NP600-1-1-198-2-1 line and NP1000-13-2-362-4-1 line about 16%, 25% shorter stem than original variety respectively through $M_6$ to $M_7$ generations. Furthermore, some characteristics of 2 lines compared to the original variety were similar or higher in weight of 1,000 seeds, number of branches per plant, number of siliqua per panicle, number of seeds per silique, oil contents, and oleic acid contents. The line with large and plump flowers selected in $M_5$ generation also showed large and dark yellow flowers through $M_6$ to $M_7$ generations. The lines with High oleic acid and low saturated fatty acid contents selected in $M_5$ generation were maintained characteristics through $M_6$ to $M_7$ generation and these useful characteristics were expected for developing a new variety for bio diesel uses.

• Journal of Radiation Industry
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• v.5 no.3
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• pp.211-220
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• 2011

This study was carried out to develop salt tolerant varities of rapeseed (Brassica napus L.) which can be grown in the high salty reclaimed land. The seeds of three varieties 'Naehan', 'Tammi', and 'Halla' were treated by proton ion beams and gamma rays with 0 to 2,000 Gy. For the selection of salt tolerant lines, emergence and survival rate, and growth characteristics of $M_2$ to $M_4$ generations were investigated in the Saemangeum reclaimed fields with the different salt concentrations. The lines with potential salt tolerance were selected in the $M_4$ generation and tested indoor for their growth characteristics. There was no significant changes in the soil pH for $M_4$ generation during growth period. However, soil EC was higher in early spring than sowing period (mid October). In $M_4$ generation test, the seeds of original and selected line showed high rates of emergence and survival, as determined one month after sowing. After wintering, however, the original varieties showed the significant reduction in the survival rate, while the selected lines showed a higher survival rate and good growth, leading to the completion of their life cycle. Consequently we selected 9 lines from $M_4$ generation with better performance in growth and yield. Soil EC was $2.8{\sim}4.3dS{\cdot}m^{-1}$ during $M_4$ generation growth period. The laboratory test of the lines selected from $M_4$ generation was made for their salt tolerance potential. The selected lines showed higher chlorophyll and proline contents than the original varieties. There was also no significant difference in the emergence rate of seed between the original and selected varieties. In 200 mM natural sea salt, the $N{\gamma}600-21-1-641$ line derived from 'Naehan' was the highest in growth rate, leaf chlorophyll and proline contents. $T{\gamma}800-20-2-461$ line derived from 'Tammi' didn't show significant difference in growth rate compared to original variety in 200 mM and withered in 250 mM like other lines as time passed. $H{\gamma}200-7-1-740$ line showed similar growth and chlorophyll content compared to its original variety.

• Korean Journal of Breeding Science
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• v.43 no.2
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• pp.145-153
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• 2011

To develop new varieties of rapeseed (Brassica napus L.), the seeds of three varieties 'Naehan', 'Tammi', and 'Halla' were treated with proton ion beams and gamma rays (0~2,000 Gy), and then the characteristics of the mutants induced were examined up to $M_5$ generation to select the lines with fixed useful traits. In $M_5$ generation, we had selected several lines that were highly fixed for some useful traits such as plant height, maturity and flower size; one line with both earlier maturity and shorter stem than wild type, one line with only earlier maturity, two lines with shorter stem, one line with large flower, and one line with chlorophyll mutation. Among them, NP600-1-1-198-2 (induced from variety 'Naehan' was treated with proton ion beams 600 Gy) was superior for its distinction from the original variety, uniformity and stability. The unique characteristics of NP600-1-1-198-2 were dark green leaves, green stem, yellow flower, and black seed coat. Its flowering date was April 14, eight days earlier than its original variety, while seed maturity date was June 16 (five days earlier) and plant height 105 cm (shorter by 10 cm). NP600-1-1-198-2 has 52 silique per panicle, 6.2 cm silique length, 23 seeds per silique, 4.2 g per 1000 seeds and 45.9% oil content. The seed oil contained 67.8% of oleic acid, 16.7% of linoleic acid and 7.3% of linolenic acid but no erucic acid.

• Journal of Veterinary Science
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• v.22 no.6
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• pp.41.1-41.14
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• 2021

Background: Our previously prepared ceftiofur (CEF) hydrochloride oily suspension shows potential wide applications for controlling swine Streptococcus suis infections, while the irrational dose has not been formulated. Objectives: The rational dose regimens of CEF oily suspension against S. suis were systematically studied using a pharmacokinetic-pharmacodynamic model method. Methods: The healthy and infected pigs were intramuscularly administered CEF hydrochloride oily suspension at a single dose of 5 mg/kg, and then the plasma and pulmonary epithelial lining fluid (PELF) were collected at different times. The minimum inhibitory concentration (MIC), minimal bactericidal concentration, mutant prevention concentration (MPC), post-antibiotic effect (PAE), and time-killing curves were determined. Subsequently, the area under the curve by the MIC (AUC_0-24h/MIC) values of desfuroylceftiofur (DFC) in the PELF was obtained by integrating in vivo pharmacokinetic data of the infected pigs and ex vivo pharmacodynamic data using the sigmoid E_max (Hill) equation. The dose was calculated based on the AUC_0-24h/MIC values for bacteriostatic action, bactericidal action, and bacterial elimination. Results: The peak concentration, the area under the concentration-time curve, and the time to peak for PELF's DFC were 24.76 ± 0.92 ㎍/mL, 811.99 ± 54.70 ㎍·h/mL, and 8.00 h in healthy pigs, and 33.04 ± 0.99 ㎍/mL, 735.85 ± 26.20 ㎍·h/mL, and 8.00 h in infected pigs, respectively. The MIC of PELF's DFC against S. suis strain was 0.25 ㎍/mL. There was strong concentration-dependent activity as determined by MPC, PAE, and the time-killing curves. The AUC_0-24h/MIC values of PELF's DFC for bacteriostatic activity, bactericidal activity, and virtual eradication of bacteria were 6.54 h, 9.69 h, and 11.49 h, respectively. Thus, a dosage regimen of 1.94 mg/kg every 72 h could be sufficient to reach bactericidal activity. Conclusions: A rational dosage regimen was recommended, and it could assist in increasing the treatment effectiveness of CEF hydrochloride oily suspension against S. Suis infections.

• Animal Bioscience
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• v.34 no.5
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• pp.801-810
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• 2021

Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.507-514
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• 2010

Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

• Journal of Dairy Science and Biotechnology
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• v.36 no.4
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• pp.208-219
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• 2018

Emergence of drug resistant strains of Salmonella enterica threatens milk processing and related dairy industries, thereby increasing the need for development of new anti-bacterials. Developments of antibacterial drugs are largely aimed to target the bacterial envelope, but screening their efficacy on bacterial envelope is laborious. This study presents a potential biosensor for envelope-specific stress in which a gfp reporter gene fused to spy gene encoding a periplasmic chaperone protein Spy (spheroplast protein y) that can sense envelope stress signals transduced by two major two-component signal transduction systems BaeSR and CpxAR in Salmonella enterica serovars Enteritidis and S. Gallinarum. Using spy-gfp operon fusions in S. Enterititis and S. Gallinarum, we found that spy transcription in both serovars was greatly induced when Salmonella cells were forming the spheroplast and were treated with ethanol or a membrane-disrupting antibiotic polymyxin B. These envelope stress-specific inductions of spy transcription were abrogated in mutant Salmonella lacking either BaeR or CpxR. Results illustrate that induction of Spy expression can be efficiently triggered by two-component signal transduction systems sensing envelope stress conditions, and thereby suggest that monitoring the spy transcription by spy-gfp operon fusions would be helpful to determine if developing antimicrobials can damage envelopes of S. Enteritidis and S. Gallinarum.