• Journal of Life Science
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• v.19 no.3
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• pp.378-383
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• 2009

Agrobacterium tumefaciens-mediated transformation procedure for the phalaenopsis orchid, established by using Protocorm-like bodies (PLBs), was aimed at the introduction of target genes into individuals with divergent genetic backgrounds. PLBs obtained from the axillary bud of a peduncle were maintained on a hyponex medium supplemented with 1 g/l of activated charcoal, 30 g/l of sucrose and 0.1 mg/l thiamine. The multiplication rate of PLBs was about 90% in case of subculture PLBs to be cut transversely into 1/3 part from top position. The PLBs were inoculated with Agrobacterium strain EHA105 harboring both $\beta$-glucuronidase (GUS) and hygromycin-resistant genes for 20 minutes after dipping treatment. Transformation efficiency was the highest with a Agrobacterium culture medium and dipping treatment of O.D. 0.8. Newly induced PLBs were put on selection medium containing 1 mg/l hygromycin for 2 months. Hygromycin-resistant phalaenopsis plants that regenerated after the selection culture of PLBs showed histochemical blue staining due to GUS. Transgene integration of the hygromycin-resistant plants was confirmed by PCR and Southern blot using GUS specific primers and probe.

• The Journal of Pediatrics of Korean Medicine
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• v.30 no.4
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• Korean Journal of Plant Tissue Culture
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• v.28 no.5
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• pp.243-248
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• 2001

This study was peformed to find out the optimal conditions for the selection of transformed cells using already established transgenic plants. Several transgenic poplar (Populus alba$\times$P giandulosa) lines carrying npt II or hpt gene as a selectable marker were tested against kanamycin or hygromycin. Two culture explants, leaf discs and nodes, were compared regarding their sensitivity to the antibiotics. When leaf discs of untransformed control plants were cultured on callus inducing media in the presence of varying levels of kanamycin or hygromycin, only those cultured on the media containing lower than 50 mg/L kanamycin or 2 mg/L hygromycin formed callus. However, much higher concentration of kanamycin was needed to suppress the growth of axillary buds of untransformed plants. On the other hand, hygromycin at the concentration of 5 mg/L effectively suppressed shoot growth of untransformed plants. Root induction from untransformed plants could also be suppressed at the concentration of 50 mg/L kanamycin or 5 mg/L hygromycin. The transgenic plants showed resistance to 100 mg/L kanamycin or 50 mg/L hygromycin in the growth of callus, shoots, and roots. Hygromycin appeared to be more efficient in selecting untransformed cells than kanamycin.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Horticultural Science & Technology
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• v.16 no.2
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• pp.239-241
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• 1998

The experiments were carried out to improve culture efficiency of rhizome and mericlone propagation through settlement of problems occurring during culture period of temperate Cymbidium species. Shooting efficiency from rhizome of C. forrestii 'Nokwoon' was improved, when cultured in $H_3P_4$ medium (Hyponex 3+peptone 4g/L) supplemented with 170mg/L $NaH_2PO_4{\cdot}H_2O$ and 0.4mg/L Thiamin e HCl, but the other varieties were not influenced to shooting efficiency by additives. Medium in which rhizome of C. nishiuchianum 'Hodukjiwha' was cultured became less browned in $H_3P_4$ medium added with 150mg/L PVP, but the other treatments of antioxidants was failed to prevent the medium browning. Re-formation of rhizome from young shoots of C. forrestii 'Sojub', 5.5cm in length occured in $H_3P_4$ enriched with 2.0 mg/L NAA and 1.0 mg/L BA under darkness, but axillary buds were elongated in the medium with 1.0 mg/L NAA and 3.0 mg/L BA under light condition. On the other hand, rhizomes from young shoot of C. forrestii 'Seosinmae' and 'Songmae', 5.5cm and 2.5cm in length respectively were reformed in 2.0 mg/L NAA and 5.0mg/L kinetin under darkness, but multishoot from young shoot were emerged in 2.0mg/L NAA and 3.0mg/L BA.

• Neonatal Medicine
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• v.18 no.2
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• pp.240-247
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• 2011

Purpose: The rationale for skin surface cultures is that bacterial colonization precedes infection and, as a result, that identification of a potential pathogen is predictive of later systemic infection in preterm infants. We aimed to analyze results of surveillance surface cultures in extremely low birth weight (ELBW) infants and seek for effective ways of performing surveillance surface cultures. Methods: We analyzed the surveillance surface cultures of 113 ELBW infants over a 4-year period. Surveillance cultures were obtained routinely from five sites: axilla, external ear canal, nasopharynx, throat (or tracheal aspirate if intubated) and anus. Each surface culture obtained during the 13 days, prior to the date of the blood culture, was compared with the blood culture obtained when sepsis was suspected. The sensitivity, specificity and positive predictive value (PPV) of the surveillance cultures were calculated among 1894 blood-surface culture pairs by surface sites, recovered organisms and interval between surveillance samples and blood cultures. Results: The overall sensitivity, specificity and PPV of surface cultures were 45.9, 22.4 and 6.8%, respectively. The PPV was highest for the throat/tracheal cultures (11.0%) and lowest for the anal cultures (2.3%). As the time of culturing progressed toward the day of blood culturing, the sensitivity and specificity of the surface cultures significantly increased. Only axillary and throat/tracheal cultures were useful in predicting the microorganisms causing sepsis. Conclusion: Surface cultures could help to predict sepsis pathogens and infection surveillance in preterm infants could be continued with a reduced number of cultured sites focusing on the axilla and throat/trachea.

• Clinical and Experimental Pediatrics
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• v.52 no.8
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• pp.898-903
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• 2009

Purpose : This study investigated the predictive factors for identifying infection-prone febrile infants younger than three months. Methods : We conducted a retrospective study of 167 infants younger than three months with an axillary temperature >$38^{\circ}C$ who were hospitalized between 2006 and 2008. If they met any of the following criteria, positive blood culture, CSF WBC ${\geq}11/mm^3$ or positive CSF culture, urinalysis WBC ${\geq}6$/HPF and positive urine culture, WBC ${\geq}6$/HPF on microscopic stool examination or positive stool culture, they were considered at high risk for severe infection. Infants with focal infection, respiratory infection or antibiotic administration prior to admission to the hospital were excluded. We evaluated the symptoms, physical examination findings, laboratory data, and the clinical course between the high risk and low risk groups for severe infection. Results : The high-risk group included 77(46.1%) infants, and the most common diagnosis was urinary tract infection (51.9%). Factors, such as male sex, ESR and CRP were statistically different between the two groups. But, a multilinear regression analysis for severe infection showed that male and ESR factors are significant. Conclusion : We did not find the distinguishing symptoms and laboratory findings for identifying severe infection-prone febrile infants younger than three months. However, the high-risk group was male and ESR-dominated, and these can possibly be used as predictive factors for severe infection.

• Journal of Korean Society of Forest Science
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• v.82 no.1
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• pp.26-33
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• 1993

In vitro shoot proliferation and rooting were tested for 2-0 seedlings of half-sib families of 4 plus oaks trees. Nodal segments having axillary buds from 37 families(16 of Quercus acutissima, 10 of Q. variabilis, 7 of Q. serrata, and 4 of Q. mongolica) were cultured on WPM(Woody Plant Medium) supplemented with 0.5 mg/l BA (6-benzyladenine) and 0.01 mg/l NAA(${\alpha}$-naphthalene acetic acid) and subcultured at 2-3 weeks of intervals fur 6 months. In vitro rooting was carried out on GD(Gresshoff and Doy) medium supplemented with 0.5mg/l IBA(indole butyric acid). The capacity for shoot proliferation and rooting was highly varied with families. Generally, white oaks(Q. serrata and Q. mongolica) showed poor response than black oaks(Q. acutissima and Q, variabilis) in shoot proliferation and rooting. Among the total of 37 families, 7 of Q. acutissima, each 2 of Q. variabilis, Q. serrata, and Q. mongolica revealed abilities for continuous shoot proliferation, and the others failed to proliferate. Rooting of the selected oak trees also greatly varied among the families. In Q. acutissima, rooting ratio ranged from 10.0%(CB 25. KG 4) to 89.8%(CB 18). Although 26.7% of KG 16 in Q. variabilis, 3.3% of JN 15 in Q. serrata were rooted, Q. mongolica was not rooted at all in this experimental conditions. No relationship between shoot growth and the rooting ability was observed. Present results suggest the possibility of large-scale micropropagation, but further studies on family differences, shoot-tip necrosis, and callusing of rooting junction are still required to develop reliable micropropagation systems.

• The Journal of Pediatrics of Korean Medicine
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• v.29 no.4
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• pp.39-66
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• 2015

Objectives The purpose of this study is to investigate the effects of Sesamum indicum extracted (SEI) on atopic dermatitis in an in-vitro and in-vivo experiment using a MC/9 murine mast cells and a NC/Nga mouse. Methods In-vitro experiment, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression were evaluated by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA and manifestations of NFAT-1, NFAT-2, c-jun, c-fos, NF-${\kappa}B$ p65 transcription factors by western blotting. In-vivo experiment, we measured WBC, Eosinophil, Neutrophil, and serum IL-5, IL-13 in NC/Nga atopic dermatitis mouse, IL-5, IL-13, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant by ELISA, the absolute cell numbers of CD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the axillary lymph node (ALN), peripheral blood mononuclear cells (PBMCs) and dorsal skin tissue, IL-5, IL-13 by Real-time PCR, the distribution of tissue inflammation and cellular infiltration by H&E and toluidine blue. Results SEI decreased IL-4, IL-5, IL-6, IL-13, GM-CSF, TNF-${\alpha}$ mRNA expression, IL-13, MIP-$1{\alpha}$ production and the expression of transcription factors including NFAT-1, c-jun, NF-${\kappa}B$ p65 in MC/9 murine mast cells. SEI orally administration decreased cell number of WBC, Eosinophil, the level of serum IgE, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, B220+CD23+ in the ALN. SEI orally administration also increased absolute cell number of CD8+/CD3+ and decreased Gr-1+/CD11b+ in PBMCs, decreased CD4+ in dorsal skin tissue, inhibited IL-5, IL-13 mRNA expression. Infiltration levels of inflammatory immune cells, mast cells and thickness of epidermis decreased in dorsal skin tissue. Conclusions SEI can regulate allergic inflammatory response suppressed the gene expression and production of cytokines that mediate allergic reactions, and will be able to be effectively utilized in the treatment of atopic dermatitis future.

• Korean Journal of Agricultural Science
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• v.14 no.2
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• pp.205-212
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• 1987

To examine the morphogenetic response, stem segments of 8 Populus spp. and 3 different explants of P. nigra var. italica were cultured on MS (Murashige and Skoog 1962) and WPM (Woody Plant Medium) medium containing various phytohormones. The results obtained were as follows: 1. Shoot regeneration and development from stem segment of 8 Populus spp. showed a quite difference according to the section and the species. All of the species of Leuce and Tacamahaca section did not form adventitious buds, while most of explants showed axillary or dormant bud elongation after 4 weeks. But P. nigra var. italica of Aigeiros section showed a successful adventitious bud formation (mean 5.4 buds per explant). 2. Leaf, petiole, and internode segment of P. nigra var. italica showed a quite differences according to media and ex plants upon the morphogenetic response. Adventitious bud formation from leaf was more abundant and readily initiated on the abaxial side than on the adaxial side. Mean number of 103 adventitious buds per explant was obtained from abaxial side of leaf segment cultured on WPM medium containing $0.2mg/{\ell}$ BAP for 5 weeks. 3. 2,4-D (2,4-dichlorophenoxy acetic acid) supplemented to media appeared to be negative upon the adventitious bud formation of P. nigra var. italica, while it promoted callus formation from all explants. Especially, NAA (${\alpha}$-naphtalene acetic acid) or NAA combination with BAP (6-benzylaminopurine) promoted root regeneration from the all explant of P. nigra var. italica in this study.