• Journal of Embryo Transfer
• /
• v.16 no.3
• /
• pp.193-201
• /
• 2001

This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

• Development and Reproduction
• /
• v.12 no.1
• /
• pp.87-95
• /
• 2008

Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.

• The Journal of Korean Academy of Prosthodontics
• /
• v.47 no.4
• /
• pp.424-434
• /
• 2009

Statement of problem: Problems such as loosening and fractures of retained screws and fracture of implant fixture have been frequently reported in implant prosthesis. Purpose: Implant has weak mechanical properties against lateral loading compared to vertical occlusal loading, and therefore, stress analysis of implant fixture depending on its material and geometric features is needed. Material and methods: Total 28 of external hexed implants were divided into 7 of 4 groups; Group A (3i, FULL $OSSEOTITE^{(R)}$Implant), Group B (Nobelbiocare, $Br{\aa}nemark$ $System^{(R)}$Mk III Groovy RP), Group C (Neobiotec, $SinusQuick^{TM}$ EB), Group D (Osstem, US-II). The type III gold alloy prostheses were fabricated using adequate UCLA gold abutments. Fixture, abutment screw, and abutment were connected and cross-sectioned vertically. Hardness test was conducted using MXT-$\alpha$. For fatigue fracture test, with MTS 810, the specimens were loaded to the extent of 60-600 N until fracture occurred. The fracture pattern of abutment screw and fixture was observed under scanning electron microscope. A comparative study of stress distribution and fracture area of abutment screw and fixture was carried out through finite element analysis Results: 1. In Vicker's hardness test of abutment screw, the highest value was measured in group A and lowest value was measured in group D. 2. In all implant groups, implant fixture fractures occurred mainly at the 3-4th fixture thread valley where tensile stress was concentrated. When the fatigue life was compared, significant difference was found between the group A, B, C and D (P<.05). 3. The fracture patterns of group B and group D showed complex failure type, a fracture behavior including transverse and longitudinal failure patterns in both fixture and abutment screw. In Group A and C, however, the transverse failure of fixture was only observed. 4. The finite element analysis infers that a fatigue crack started at the fixture surface. Conclusion: The maximum tensile stress was found in the implant fixture at the level of cortical bone. The fatigue fracture occurred when the dead space of implant fixture coincides with jig surface where the maximum tensile stress was generated. To increase implant durability, prevention of surrounding bone resorption is important. However, if the bone resorption progresses to the level of dead space, the frequency of implant fracture would increase. Thus, proper management is needed.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.28 no.5
• /
• pp.512-519
• /
• 2014

This study investegated the effect of Liriope platyphylla (LP) on allergic reactions and its mechanism of action. We investigated the effect of LP on Evans Blue (EB) extravasation induced by anti-dinitrophenyl (DNP)-IgE in rats. We tested whether the ethanol extract of LP reduced ear skin thickness and historical changes induced by topical application of 2,4-dinitrofluorobenzene (DNFB) to ears of mice. We evaluated compound 48/80-induced release of histamine in rats peritoneal mast cell (RPMCs). We also investigated the regulatory effect of LP on the level of inflammatory mediators in PMACI-induced human mast cell (HMC-1); cytokine IL-6, IL-8, TNF-${\alpha}$ in HMC-1, MAPKs (ERK, JNK and p38) in HMC-1. The ethanol extract of LP (81.3 mg/100 g body weight) significantly inhibited the PCA reaction compared with the control (P < 0.05). However, LP did not prevent topical applications of DNFB-induced ear skin thickening and histological changes. In RPMCs, histamine release induced by compound 48/80 was significantly attenuated by LP at $100{\mu}g/ml$ (P < 0.05). LP extract ($100{\mu}g/ml$) significantly reduced the PMACI-induced IL-6, IL-8, and TNF-${\alpha}$ secretion via inhibition of ERK phosphorylation in HMC-1. In conclusion, the ethanol extract of LP inhibited mast cell-derived, immediate-type allergic reactions, and the result suggest the potential of LP for preventing allergic inflammatory disorders.

• Food Science and Biotechnology
• /
• v.17 no.1
• /
• pp.1-7
• /
• 2008

Soybeans with brown, black, and yellow seed coats were compared to total phenolic contents and antioxidant activities including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals. Also, 3 seed coats were examined for inhibitory activities on tyrosinase and lipoxygenase-1 on the basis of spectrophotometric and polarographic methods. Among seed coat extracts, 80% methanol extract of brown soybean seed coat showed the highest total phenolic contents ($68.9{\pm}3.29\;mg$ GAE/g) as well as exhibited potent scavenging effects on the DPPH ($IC_{50}=4.3\;{\mu}g/mL$) and ABTS ($IC_{50}=3.7\;{\mu}g/mL$) radicals. In a polarographic experiment, this extract was potentially inhibited the oxidation of L-tyrosine and L-3,4-dihydroxy-phenylalanin (L-DOPA) catalyzed by mushroom tyrosinase with $IC_{50}$ values of 12.4 and $63.7\;{\mu}g/mL$, respectively. It was also detected inhibition of the tyrosinase catalyzed oxidation of L-DOPA with an $IC_{50}$ value of 120.3 mg/mL in UV spectrophotometric experiment. In addition, this extract inhibited the linoleic acid peroxidation catalyzed by lipoxygenase-1 with an $IC_{50}$ value of $4.0\;{\mu}g/mL$. These results suggest that brown soybean may possess more beneficial effect on human health than black and yellow soybeans.

• Clinical and Experimental Reproductive Medicine
• /
• v.31 no.1
• /
• pp.67-74
• /
• 2004

Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

• Korean journal of communication and information
• /
• v.53
• /
• pp.53-72
• /
• 2011

has become the representative program of EBS after first airing in 2004. After seeing as a media text, this study was fulfilled on the basis of Bill Nichols' "Rhetoric" discussion, in order to analyze the structural method and social meaning of the program. As a result of the study, we were first able to see that was taking form as a documentary to make a constructive statement on current affairs related to Korean society, by mobilizing 'knowledge' related to these issues. Second, the "rhetoric" plays an important role in the program's process to generate meaning, and despite the text being relatively short, the rhetorical process undergone for the generation of meaning has been fully reflected. Moreover, the program was successfully structuring a new method of generating meaning by enthusiastically raising questions and choosing a unique method of vision mix. Third, by looking at the study's results in-depth, we can see that was creating a ideological place for open discussion, which was providing viewers with the satisfaction of overcoming our society's dominating ideology and forming new ones. In that sense, the easily accessible 'knowledge'that surrounds us holds somewhat of a political meaning. On top of that, it can be seen that the program especially, is a media text that holds a cultural political meaning, which is an important factor to consider in the Korean broadcasting industry. In conclusion, we found out that this program uses 'knowledge' to see the possibility of 'knowledge'journalism by making constructive statements through a 'rhetorical' process.

• Reproductive and Developmental Biology
• /
• v.35 no.1
• /
• pp.9-15
• /
• 2011

The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.30 no.6
• /
• pp.504-508
• /
• 2004

Surgical process of oral and maxillofacial area as well as dental treatment are stressful situations to the patients. It is well known that serum level of stress hormones including adrenocorticotropic hormone(ACTH) and cortisol increase when the body is exposed to stress. However, there have been few studies on demographic and clinical factors related with stress. Therefore, the purpose of this study was to supply data to cope with stress more comprehensively and efficiently by analyzing the effect of factors related with stress in addition to surgical procedure. Prospective cohort study method was selected. Serum levels of ACTH and cortisol were measured by sampling bloods pre-operatively and post-operatively from 45 patients who had been operated at the Bundang Jesaeng Hospital department of oral and maxillofacial surgery. To evaluate factors associated with stress, patients were classified according to gender, age, method of payment(insurance or self), experiences of operation, kind of operations(expected operations or unexpected operations). Relative risk was calculated to assess relationships between changes of serum level of ACTH and cortisol and factors related with stress, whereas Chi-square analysis was executed to evaluate statistical significance. With regard to serum level of ACTH, relative risk was 1.3 in the group of the patients who were less than 40 years old. With regard to serum level of cortisol, relative risk was 1.8 for women compared with men, 1.4 in the group of the patients who were less than 40 years old and 1.6 in the group of the patients who had not experienced any other operations. In addition to surgical procedure, factors related with stress included gender, age, method of payments, experiences of operation and kind of operations. Therefore, we should provide comprehensive schemes to reduce stress of the patients going through oral and maxillofacial surgery.

• Journal of Applied Biological Chemistry
• /
• v.51 no.1
• /
• pp.20-27
• /
• 2008

The contents of lignan, protein, and oil of nine Sesamum indicum cultivars were analyzed. To determine the lignan contents, sesamin 1 and sesamolin 2 were isolated from the ethylacetate extracts of the Suwon cultivar, and the structures were fully characterized by spectral and physical methods. The seeds of nine cultivars were screened for two lignans, which were determined by HPLC using a $C_{18}$ reversed phase column coupled with a photodiode array detector. Suwon cultivar showed the highest concentration (1: $6.24{\pm}0.04$ and 2: $3.58{\pm}0.01\;mg/g$), whereas Soonheuk displayed the lowest (1: $0.91{\pm}0.01$ and 2: $0.73{\pm}0.01\;mg/g$). The average content ratio of sesamin 1 (3.64 mg/g) was significantly higher than that of sesamolin 2 (2.57 mg/g). The protein content ranged from $21.52{\pm}0.35$ to $31.22{\pm}0.25%$, Suwon containing the highest level and Soonheuk had the lowest. Kwangbaek showed the highest oil level ($49.84{\pm}0.40%$), while Soonheuk exhibited the lowest ($42.52{\pm}0.05%$). Sesamin 1 exhibited a stronger radical-scavenging activity in the ABTS ($IC_{50}:\;63.2{\pm}2.4\;{\mu}M$) than its DPPH radical-scavenging activity ($IC_{50}>200\;{\mu}M$). These results lead to the conclusion that lignan content is affected by protein and oil contents. Concentration of the chemical components in the cultivars could be a key factor in the selection process of a high quality species.