• Clinical and Experimental Pediatrics
• /
• v.45 no.2
• /
• pp.247-255
• /
• 2002

Purpose : This study was undertaken to obtain basic data about the megakaryocyte colony formation of fetal liver cells by using immunocytochemical staining and ex vivo culture with growth factors. Methods : The mononuclear cells were isolated from fetal liver and bone marrow with idiopathic thrombocytopenic purpura(ITP) and pancytopenia. These mononuclear cells were cultured in $MegaCult^{TM}-C$(Stem Cell Tech, Canada) media in the presence of growth factors and CFU-Megakaryocyte( CFU-Mk) colonies were counted on day 12. The expansion of CD34+ and CD41+ cell was analyzed by flow cytometry after 5 days incubation using flask culture. Results : The numbers of CFU-Mk colonies of mononuclear cells obtained from fetal liver in the 11th week gestational age were more than those in the 19th week specimens; growth factors could not enhance the colony expansion in all cases. Total numbers of CFU-Mk colony of fetal liver cells were higher than bone marrow from ITP or pancytopenia groups. The numbers of pure or large CFU-Mk colonies of fetal liver cells were also higher than bone marrow specimens. The rate of CD34+ cell expression of fetal liver was increased after flask culture and the enhancement effect of epression was seen only in cases which added thrombopoietin. The rate of CD41+ cell expression of fetal liver was increased after incubation, but the enhancement effect of growth factors was unclear. Conclusion : This study revealed good results about the megakaryocyte colony assay of fetal liver mononuclear cells using $MegaCult^{TM}-C$ media. This study suggests that the fetal liver could be a good source of megakaryocytic progenitor cells for clinical application in hematopoietic stem cell transplantation.

• Korean Journal of Plant Resources
• /
• v.23 no.5
• /
• pp.415-422
• /
• 2010

Red ginseng(RG, steamed and dried root of Panax ginseng C. A. Meyer, family Araliaceae) and fermented red ginseng(FRG, fermented red ginseng by yeast and lactic acid bacteria) are known to show different pharmacological effects by changed composition of saponins through fermentation. We examined the effects of RG and FRG on $\beta$-hexosaminidase secretion, ICAM-1 expression, the mitogen-induced proliferation of lymphocyte from mice in ex vivo systems and HaCaT cell(keratinocyte) proliferation to compare the anti-allergic and anti-inflammatory effects between both groups. RG groups showed inhibition of $\beta$-hexosaminidase secretion and ICAM-1 expression at $1{\mu}g/ml$, $10{\mu}g/ml$ and the same effects were observed at all concentrations in FRG groups. In our study, RG increased LPS-induced B cell proliferation at $1{\mu}g/ml$ and ConA-induced B cell proliferation at $100\;{\mu}g/ml$ but FRG decreased LPS- and ConA-induced lymphocytes at $100\;{\mu}g/ml$. We showed that FRG increased the proliferation of HaCaT at 1, $10{\mu}g/ml$ but not by RG. These findings suggest that RG and FRG might have anti-inflammatory and anti-allergic effects, which can be needed to proper clinical concentration to applied to various allergic diseases and inflammation.

• Korean Journal of Microbiology
• /
• v.43 no.4
• /
• pp.298-303
• /
• 2007

Antithrombotic activities of water extract of cheongkookjang and cheongkookjang fermented with green tea or mugwort were evaluated on some antithrombosis related activities in vitro and thrombotic death inhibition in vivo. Cheongkookjang made of white soybean (Glycine max) or black small soybean (Rhynchosia nulubilis) showed potent antioxidative activities. Addition of green tea or mugwort during cheongkookjang fermentation increased the antioxidative activity, cheongkookjang with green tea showed more drastic increase compared with cheongkookjang with mugwort. Nitrite scavenging effects of the cheongkookjang extracts were prominent but the addition of green tea or mugwort seldom increased the scavenging effects. All the cheongkookjang extracts showed strong inhibitory activities on platelet aggregation. The inhibitory activities of cheongkookjang were increased considerably by addition of green tea or mugwort even with low concentration. Plasmin unit as fibrinolytic activity was not affected considerably by addition of green tea. Addition of mugwort decreased the activity transiently at low concentration ($0.3{\sim}1.0%$) but increased again slowly at higher concentration ($1{\sim}3%$). In vitro thrombotic death inhibition test, the antithrombotic activity of cheongkookjang made of black small bean with green tea was higher by about 1.5 times compared to that without green tea. As results, cheongkookjang might inhibit antithrombosis not only by fibrinolytic action but also by inhibition of platelet aggregation and antioxidative action. The addition of functional materials such as green tea or mugwort could increase the antithrombotic function, even at low concentration.

• Korean Journal of Poultry Science
• /
• v.28 no.2
• /
• pp.135-142
• /
• 2001

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

• Tuberculosis and Respiratory Diseases
• /
• v.54 no.5
• /
• pp.542-550
• /
• 2003

Background : Synthetic glucocorticoids are widely used in many chronic inflammatory diseases because of their excellent anti-inflammatory activity. Enhancing the transcription of $I{\kappa}B$ and preventing activated NF-${\kappa}B$ from binding to ${\kappa}B$ sites are thought to be the underlying mechanisms. But these data are largely derived from in vitro studies using cell lines. In this study, after administrating a steroid to volunteers, we evaluated the effect on the NF-${\kappa}B$ system. Methods : Prednisolone(0.5mg/kg/d) was orally administered to 5 healthy volunteers for 7 days. Before and after the administration, we sampled their peripheral blood monocytes, and performed western blot analysis both with stimulation, using IL-$1{\beta}$, LPS, TNF, and without stimulation(baseline). We also performed EMSA after stimulation with LPS. Results : After ingestion of the steroid, baseline expressions of $I{\kappa}B{\alpha}$ were increased in two of the subjects, while suppressed degradations of $I{\kappa}B{\alpha}$ to stimulations were observed in all five. In addition, the binding capacity of NF-${\kappa}B$ after the administration was decreased. Conclusion : Steroid plays such roles as enhancing the transcription of $I{\kappa}B{\alpha}$, suppressing the DNA binding capacity of NF-${\kappa}B$, and suppressing the degradation of $I{\kappa}B{\alpha}$.

• Restorative Dentistry and Endodontics
• /
• v.41 no.2
• /
• pp.91-97
• /
• 2016

Objectives: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human ${\beta}$-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. Materials and Methods: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and $300{\mu}g/mL$), CH ($100{\mu}g/mL$), and Nys ($20{\mu}g/mL$) for 7 days at $37^{\circ}C$. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. Results: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (${\geq}100{\mu}g/mL$). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (${\geq}100{\mu}g/mL$) showed significant fungicidal activity compared to CH group (p < 0.05). Conclusions: Synthetic HBD3-C15 peptide (${\geq}100{\mu}g/mL$) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

• Korean Journal of Optics and Photonics
• /
• v.31 no.1
• /
• pp.1-6
• /
• 2020

The chest wall, an organ directly affected by environmental particles through respiration, consists of ribs, a pleural layer and intercostal muscles. To diagnose early and treat disease in this body part, it is important to visualize the details of the chest wall, but the structure of the pleural layer cannot be seen by chest computed tomography or ultrasound. On the other hand, optical coherence tomography (OCT), with a high spatial resolution, is suited to observe pleural-layer response to talc, one of the fine materials. However, intensity-based OCT is weak in providing information to distinguish the detailed structure of the chest wall, and cannot distinguish the reaction of the pleural layer from the change in the muscle by the talc. Polarization-sensitive OCT (PS-OCT) takes advantage of the fact that specific tissues like muscle, which have optical birefringence, change the backscattered light's polarization state. Moreover, the birefringence of muscle associated with the arrangement of myofilaments indicates the muscle's condition, by measuring retardation change. The PS-OCT image is interpreted from three major perspectives for talc-exposure chest-wall imaging: a thickened pleural layer, a separation between pleural layer and muscle, and a phase-retardation measurement around lesions. In this paper, a rabbit chest wall after talc pleurodesis is investigated by PS-OCT. The PS-OCT images visualize the pleural layer and muscle, respectively, and this system shows different birefringence of normal and damaged lesions. Also, an analyisis based on phase-retardation slope supports results from the PS-OCT image and histology.

• Korean Journal of Plant Resources
• /
• v.23 no.2
• /
• pp.157-164
• /
• 2010

Rosa rugosa has been used as a folk medicine with various pharmacological properties for a long time in Asia. We investigated effects of fructus extracts of Rosa rugosa (RRF) and semen extracts of this herb (RRS) on bone forming cells (osteoblastic and pre-osteoclastic cells) to evaluate the pharmacological possibilities in a variety of bone-related disease. RRF showed significant effect on proliferation of osteoblastic cells in dose-dependent manners at 72 hrs and $100\;{\mu}g/m{\ell}$ of RRS was effective at 48 and 72 hrs. RRF and RRS did not decreased production of TNF-$\alpha$ but NO by pre-osteoclastic cells under inflammation circumstance indeced by LPS. We also investigated the effects of RRF and RRS on the mitogen-induced lymphocyte proliferation in the old and young mice in ex vivo systems. RRF and RRS significantly enhanced proliferative effects of untreated and ConA-treated splenocytes from the old and young mice. But, RRS at $500\;{\mu}g/m{\ell}$ increased LPS-induced TNF-$\alpha$ production in pre-osteoclastic cells and reduced LPS-stimulated lymphoblastogenesis in the old and young at $1000\;{\mu}g/m{\ell}$. These results indicate that RRF has beneficial effects on osteoarthritis and give further possibilities for the immunomodulating effects not only in old that has more frequent bone related diseases but also in young.

• Korean Journal of Food Science and Technology
• /
• v.51 no.4
• /
• pp.379-392
• /
• 2019

The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

• The Korean Journal of Nuclear Medicine
• /
• v.39 no.6
• /
• pp.421-429
• /
• 2005

Purpose: It has been postulated that dopamine release in the striatum underlies the reinforcing properties of nicotine. Substantial evidence in the animal studies demonstrates that nicotine interacts with dopaminergic neuron and regulates the activation of the dopaminergic system. The aim of this study was to visualize the dopamine release by smoking in human brain using PET scan with $[^{11}C]raclopride$. Materials and Methods: Five male non-smokers or ex-smokers with an abstinence period longer than 1 year (mean age of $24.4{\pm}1.7$ years) were enrolled in this study $[^{11}C]raclopride$, a dopamine D2 receptor radioligand, was administrated with bolus-plus-constant infusion. Dynamic PET was performed during 120 minutes ($3{\times}20s,\;2{\times}60s,\;2{\times}120s,\;1{\times}180s\;and\;22{\times}300s$). following the 50 minute-scanning, subjects smoked a cigarette containing 1 mg of nicotine while in the scanner. Blood samples for the measurement of plasma nicotine level were collected at 0, 5, 10, 15, 20, 25, 30, 45, 60, and 90 minute after smoking. Regions for striatal structures were drawn on the coronal summed PET images guided with co-registered MRI. Binding potential, calculated as (striatal-cerebellar)/cerebellar activity, was measured under equilibrium condition at baseline and smoking session. Results: The mean decrease in binding potential of $[^{11}C]raclopride$ between the baseline and smoking in caudate head, anterior putamen and ventral striatum was 4.7%, 4.0% and 7.8%, respectively. This indicated the striatal dopamine release by smoking. Of these, the reduction in binding potential in the ventral striatum was significantly correlated with the cumulated plasma level of the nicotine (Spearman's rho=0.9, p=0.04). Conclusion: These data demonstrate that in vivo imaging with $[^{11}C]raclopride$ PET could measure nicotine-induced dopamine release in the human brain, which has a significant positive correlation with the amount or nicotine administered bt smoking.