• Korean Journal of Animal Reproduction
• /
• v.25 no.1
• /
• pp.71-77
• /
• 2001

This experiment was designed to improve the production efficiency of germ-line chimeric mice from phospholipase C (PLC)-$\beta$3 or peroxiredoxin (Prx) E -targeted ($\Delta$) ES cells by the investigating the manipulation conditions and characteristics of Jl ES cells. Four targeting clones were isolated to investigate the karyotypical and morphological stability prior to injection. All clones ($\Delta$PLC$\beta$-3 C3, $\Delta$Prx II C3, C10 and I5) showed more than 80% euploidism, however, most of $\Delta$PLC$\beta$-3 C3 clones were extensively differentiated compared to the other clones. Nine of 13 $\Delta$Prx II chimeras appeared to have at least 80% chimerism, whereas $\Delta$PLC$\beta$-3 C3 chimeras had 20% chimerism at most. Therefore, the morphological stability of ES cells under stable euploidism might mainly affect the production rate of high-coat chimeric mice. To increase the collection rate of injectable blastocysts (IBs), 5 to 10 week -aged C57BL/6J female mice were sacrificed at 3.5 days post-coitum. Ten week-aged mice were the most optimal IB donors by showing the highest collection rate (2.94/mouse) of injectable blastocysts without increase of non-injectable embryos (0.29/mouse). Foster mothers might be another factor because ICR x C57BL/6J F1 foster mother showed more increased productivity in litter size (2.8 vs. 5.6) and chimera (0 vs. 35.3%) than those of ICR foster mothers. In conclusion, the efficient production of germ-line chimeras mainly depends on the maintenance of ES cell morphology during targeting procedure, and the establishment of manipulation conditions might be a key point to maximize it.

• Journal of Microbiology
• /
• v.44 no.1
• /
• pp.64-71
• /
• 2006

To investigate the effects of the nucleotide sequences in Shine-Dalgarno (SD) and the spacer region (SD-ATG) on bovine growth hormone (bGH) gene expression, the expression vectors under the control of the T7 promoter (pT7-7 vector) were constructed using bGH derivatives (bGH1 & bGH14) which have different 5'-coding regions and were induced in E. coli BL21 (DE3). Oligonucleotides containing random SD sequences and a spacer region were chemically synthesized and the distance between the SD region and the initiation codon were fixed to nine bases in length. The oligonucleotides were annealed and fused to the bGH1 and bGH14 cDNA, respectively. When the bGH gene was induced with IPTG in E. coli BL21(DE3), some clones containing only bGH14 cDNA produced considerable levels of bGH in the range of $6.9\%\;to\;8.5\%$ of total cell proteins by SDS-PAGE and Western blot. Otherwise, the bGH was not detected in any clones with bGH1 cDNA. Accordingly, the nucleotide sequences of SD and the spacer region affect on bGH expression indicates that the sequences sufficiently destabilize the mRNA secondary structure of the bGH14 gene. When the free energy was calculated from the transcription initiation site to the +51 nucleotide of bGH cDNA using a program of nucleic acid folding and hybridization prediction, the constructs with values below -26.3 kcal/mole (toward minus direction) were not expressed. The constructs with the original sequence of bGH cDNA also did not show any expression, regardless of the free energy values. Thus, the disruption of the mRNA secondary structure may be a major factor regulating bGH expression in the translation initiation process. Accordingly, the first stem-loop among two secondary structures present in the 5'-end region of the bGH gene should be disrupted for the effective expression of bGH.

• Clinical and Experimental Pediatrics
• /
• v.56 no.11
• /
• pp.490-495
• /
• 2013

Purpose: The use of cyclosporine and mini-dose methotrexate (MTX) is a common strategy for graftversus- host disease (GVHD) prophylaxis in allogeneic transplants. We investigated whether patients who receive fewer than the planned MTX doses are at increased risk for GVHD. Methods: The study cohort included 103 patients who received allogeneic transplants at the Department of Pediatrics of The Catholic University of Korea College of Medicine, from January 2010 to December 2011. MTX was administered on days 1, 3, 6, and 11 after transplant at a dose of 5 $mg/m^2$ each. Within the cohort, 76 patients (74%) received all 4 doses of MTX [MTX(4) group], while 27 patients (26%) received 0-3 doses [MTX(0-3) group]. Results: Although there was no difference in neutrophil engraftment between the 2 groups, platelet engraftment was significantly faster in the MTX(4) group (median, 15 days), compared to the MTX(0- 3) group (median, 25 days; P =0.034). The incidence of grades II-IV acute GVHD was not different between the MTX(4) and MTX(0-3) groups (P =0.417). In the multivariate study, human leukocyte antigen mismatch was the most significant factor causing grades II-IV acute GVHD (P =0.002), followed by female donor to male recipient transplant (P =0.034). No difference was found between the MTX(4) and MTX (0-3) groups regarding grades III-IV acute GVHD, chronic GVHD, and disease-free survival. Conclusion: Our results indicate that deviations from the full dose schedule of MTX for GVHD prophylaxis do not lead to increased incidence of either acute or chronic GVHD.

• Journal of radiological science and technology
• /
• v.21 no.1
• /
• pp.79-87
• /
• 1998

The total-bodies of 10 week-old Sprague-Dawley rats were irradiated with single doses 4.5 and 7.5 Gy, respectively. The effects on plasma and sciatic nerve platelet-derived growth factor(PDGF) concentrations and sciatic nerve PDGF ${\alpha}$ -and ${\beta}$ -receptors densities were examined up to 10 days post-treatment. There was no consistent significant variation in the plasma and sciatic nerve PDGF concentrations in time over the period of study between 4.5 and 7.5 Gy groups. Plasma PDGF concentrations were significantly reduced to 58% of control values between 5 and 10 days with 4.5 Gy and to 51% of control values as percentage of control values between 5 and 10 days with 7.5 Gy after irradiation, respectively(p<0.05). Sciatic nerve PDGF concentrations were increased to 118% of control values at 1 day with 4.5 Gy and to 130% of control values at 1 day with 7.5 Gy after irradiation, respectively(p>0.05). After irradiation, the levels of PDGF ${\alpha}$ -receptor protein density were reduced to 33% of control values at 2 days with 4.5 Gy and to 50% at 2 days with 7.5 Gy, while the levels of PDGF ${\beta}$-receptor protein density were reduced to maximally 26% of control values at 2 days with 4.5 Gy and to 27% at 2 days with 7.5 Gy, respectively, but both initial decreased levels of those were increased subsequently after 2 days following irradiation. These results suggest that the radiation-induced alteration of plasma and sciatic nerve PDGF concentrations, and sciatic nerve PDGF ${\alpha}$ -and ${\beta}$ -receptors densities may be involved in the pathogenesis of bone marrow stem cell and peripheral neuron damages.

• Korean Journal of Plant Resources
• /
• v.26 no.4
• /
• pp.441-449
• /
• 2013

This study was conducted to select some useful plants as functional material candidates. A total of 38 plants were preliminarily screened for the anti-inflammatory and antioxidant activities. The preliminarily selected 8 plants were further investigated to verify the in vitro inhibitory effect on inflammation and oxidative stress. Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) inhibited the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Eupatorium japonicum (leaf) suppressed the expression of cyclooxygenase-2 (COX-2), whereas Boehmeria platanifolia (root) and Prunus yedoensis (branch) inhibited the transcription of nuclear factor-kappa B (NF-${\kappa}B$). Treatment with the extracts ($2.5{\sim}20{\mu}g/ml$) of Abutilon theophrasti (leaf, flower/seed) and Hemistepta lyrata (stem) did not show toxicity on RAW 264.7 cell proliferation, but treatment with $2.5{\mu}g/ml$ of Boehmeria platanifolia (root) exhibited cell toxicity. Carpinus coreana (branch) and Prunus yedoensis (branch) showed potent scavenging activities on peroxynitrite. Akebia quinata (flower), Carpinus coreana (branch), and Prunus yedoensis (branch) effectively inhibited reactive oxygen species (ROS). Abutilon theophrasti (leaf), Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) exhibited strong inhibitory capacity with regard to nitric oxide (NO) production. The results suggested that Abutilon theophrasti (leaf) has in vitro anti-inflammatory and antioxidant activities, and that is a useful functional material candidate.

• Journal of Life Science
• /
• v.20 no.8
• /
• pp.1221-1229
• /
• 2010

Nerium indicum, an India-Pakistan-originated shrub belonging to the oleander family, is reported to possess many pharmacological activities including cardiac muscle stimulation, and anti-diabetes, anti-angiogenesis, anti-cancer and neuro-protective activities. However, the anti-inflammatory properties of N. indicum were unclear. In this study, we investigated the effects of ethanol extract of the N. indicum leaf and stem (ENIL and ENIS) on the expression of anti-inflammatory mediators in U937 human pre-monocytic cell models. In U937 cells stimulated with phorbol 12-myristate-13-acetate (PMA), pre-treatment with ENIS significantly inhibited the expression of both cyclooxygenase-2 (COX-2) mRNA and protein, which are associated with inhibition of the release of prostaglandin $E_2\;(PGE_2)$, whereas the inhibitory effects appeared weakly in ENIL. Moreover, ENIS significantly attenuated PMA-induced IkappaB ($I{\kappa}B$) degradation and suppressed elevated nuclear factor kappa B (NF-${\kappa}B$) nuclear translocation. Taken together, these findings provide important new insights that N. indicum exhibits anti-inflammatory properties by suppressing the transcription of pro-inflammatory cytokine genes through the NF-kB signaling pathway.

• Journal of The Korean Association For Science Education
• /
• v.36 no.5
• /
• pp.757-768
• /
• 2016

This study developed an SSI (Socio-Scientific Issue) discussion program that applies a creative technique called six thinking hats, and then investigated the differences in argumentation patterns and effects on the decision-making abilities of each character feature of students between SNS debate and existing face to face debate. There were three SSI themes - Designer Babies, embryonic stem cell study, and legitimacy of abortion. Students were divided into two groups, the debate group using SNS and face to face debate group. The character patterns of students were divided to 'extraversion,' 'agreeableness,' and 'conscientiousness' through test sheets for character features for each student. Both groups were educated for creative discussion methods using six thinking hats and then, the class progressed. As a result of analyzing argumentation patterns used in SNS debate and face to face debate, the most used argumentation pattern was the "cause pattern." But comparing to face to face debate, other patterns (mark, inference, authority, motive) were also used in SNS debate. The study analyzed three factors of decision-making ability for each character feature of students such as complexity, perspectives, and inquiry. As a result, for 'complexity' factor, there was a significant difference between SNS debate group and face to face debate group only in the student group of Agreeableness. For 'perspectives' factor, there were significant differences between SNS debate group and face to face debate group in all three characters. Finally, for inquiry, there were no significant differences between SNS debate group and face to face debate group in all three characters. Accordingly it would be necessary to apply SNS debate using the six thinking hats in SSI education to enhance perspectives.

• Pediatric Infection and Vaccine
• /
• v.23 no.1
• /
• pp.31-39
• /
• 2016

Purpose: The aim of this study was to investigate the prognostic factors for Pneumocystis jirovecii pneumonia (PCP) and to evaluate the influence of PCP prophylaxis in pediatric patients. Methods: From January 2002 to April 2015, patients aged <18 years with a diagnosis of confirmed PCP at our institute were reviewed retrospectively. Clinical characteristics and outcomes were compared according to the groups with or without PCP prophylaxis. Risk factors associated with PCP-related death were analyzed by logistic regression analysis. Results: During study period, a total of 24 patients were diagnosed with PCP by immunofluorescence assay and/or PCR. The median age of the patients was 5 years (range, 3 months-18 years) and 23 (96%) had immunocompromised conditions including hematologic disorders with or without hematopoietic stem cell transplantation (n=15), solid organ transplantation (n=4), and primary immune deficiency (n=4). Most common presenting symptoms were tachypnea and cough (92%, each). At the time of diagnosis, 79% (19/24) and 25% (6/24) suffered from respiratory failure and multi-organ dysfunction syndrome (MODS), respectively. Mechanical ventilation was required in 8 (33%) patients and 5 (21%) patients died of PCP. Multivariate analysis showed that MODS at initial presentation was an indicator of poor prognosis (OR, 17.1 [95% CI 1.13-257.67]; P=0.04). Compared to the patients without PCP prophylaxis, the frequency of MODS at diagnosis, need for mechanical ventilation and length of hospital days were significantly less common in the children who received PCP prophylaxis. Conclusions: MODS at presentation was a significant predictor for poor outcome and PCP prophylaxis could alleviate the clinical courses of pediatric PCP. Prospective study will be mandatory to determine the risk factors for development and deterioration of PCP in children.