• Title/Summary/Keyword: In vitro Development

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Improvement of Seed Germination in a Spontaneous Autotetraploid of Poncirus and Chlorophyll Fluorescence of Seedlings in Salt Stress (동질 사배체 탱자의 종자 발아 증진과 염류 과잉에 따른 엽록소 형광 반응)

  • Chae, Chi Won;Yun, Su Hyun;Park, Jae Ho;Kim, Min Ju;Han, Seung Gab;Kang, Seok Beom;Koh, Sang Wook;Han, Sang Heon
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1079-1087
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    • 2013
  • Speed germination success and robust vegetative growth of citrus rootstock through improved sowing methods and fertilizer inputs offer the usage of root system for the citrus. The current study evaluated the influence of seed coat removal and different fertilizer concentrations on plant germination and plant growth of spontaneous rootstock siblings. Decoated and coated seeds of diploid and tetraploid plants were sown in tubes. Commercial fertilizer concentrations of 0, 2, 4, 6, 8 and $10g{\cdot}l^{-1}$ were added. The experimental layout followed a randomized block $2{\times}6$ factorial design (seed coat removal ${\times}$ fertilizer concentration) for each rootstock. Fertilizer concentrations were 0, 10, 20 and $30g{\cdot}l^{-1}$ of the fertilizer for the resistance of the strength on the salt level. The germination rate of seeds without testa sown in vitro was improved (67-80%) compared to that of nontreated seeds. The eventual tree height of the seeds without testa in the diploid group was increased due to higher fertilization compared to that in the nontreated group. The removal of seed testa promoted the seed germination of both diploid and tetraploid trifoliate orange and resulted in greater height. Their vegetative development was also increased due to the increased fertilization of the rootstock. The Fv/Fm value for the diploid plants was 0.4 and 0.8 for the tetraploid ones under salt stress after 11 days of treatment. The removal of seed testa may improve the seed germination of trifoliate orange. Tetraploid trifoliate orange appears to possess resistance to salt stress compared to the diploid variety.

DEVELOPMENT OF HIGH SENSITIVE MODEL OF CARIES ACTIVITY TEST FOR EARLY DIAGNOSIS OF DENTAL CARIES (치아우식증의 조기진단을 위한 고감도 우식활성검사 모형개발)

  • Lee, Sang-Ho;Lee, Chang-Seop
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.169-179
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    • 2000
  • The purpose of this study is to develop the system which convert the optical difference of teeth texture between intact enamel and incipient caries lesion into shade difference by laser fluorescence and to develop new and simple caries activity test using laser fluorescence. The experimental design of this study consists of three parts. In first part, a new method for the in vitro assessment of changes in initial enamel caries lesion of Bovine teeth using laser fluorescence is tested. In second part, in vivo assessment undertaken. Number of teeth which showed incipient carious lesion on buccal surface examined by laser fluorescence was compared with the caries activity test of $Cariescreen^{(R)}$ test and other oral environmental test of dDfFtT. In third part, new caries activity test measured by laser fluorescence was developed on the basis of above results and evaluated the sensitivity, specificity, and diagnostic power. Optical density measured by laser fluorescence was increased as increasing the depth of incipient carious lesion and showed high correlation$(\gamma=0.7015)$ with lesion depth. Optical density showed direct proportion to lesion depth. Linear equation was obtained between the optical density and the lesion depth by regression analysis. The result of caries activity test with laser fluorescence showed high correlation with those of $Cariescreen^{(R)}$ test and dDfFtT examination. Caries activity test with laser fluorescence showed 48% of sensitivity, 52% of specificity, and 45% of diagnostic power on the basis of dDfFtT examination, and also showed 48% of sensitivity, 51% of specificity, and 36% of diagnostic power on the basis of $Cariescreen^{(R)}$ test. In regard above result, caries activity test with laser fluorescence considered to be reliable for caries activity test compared with other oral environmental test. and it was also considered to be practical because it would be simple, inexpensive, and time saving method.

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Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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Discontinuous Percoll Gradients Enrich X-Bearing Porcine Sperms and Female Embryos (불연속 Percoll 원심분리에 의한 돼지 X-정자와 자성배아에 관한 연구)

  • Shim, Dae-Yong;Yoo, Seong-Jin;Kang, Han-Seung;Yoo, Jeong-Min;Lee, Chae-Kwan;Kang, Sung-Goo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.47-52
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    • 2001
  • Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

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Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Development of Pharmaceutical Dosage Forms with Biphasic Drug Release using Double-Melt Extrusion Technology (이중 고온용융 압출 성형된 이중 방출능을 가지는 제형의 개발)

  • Kim, Dong-Wook;Kang, Chin-Yang;Kang, Changmin;Park, Jun-Bom
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.9
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    • pp.228-234
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    • 2016
  • The aim of this study was to develop pharmaceutical dosage forms with a bi-phasic drug using a double extrusion approach. Hot melt extrusion was performed using a co-rotating twin-screw extruder. The. 1st melt extrusion was performed using polymer with a relatively higher Tg, such as HPMC and the 2nd melt extrudate was obtained using the 1st extrudate and polymers with a lower Tg, such as HPMC-AS and PEO. In addition, the formulation with all the content in the same proportion as the double extudate was produced using single extrusion for comparison. Physical characterization was performed on the formulations employing differential scanning calorimetry (DSC). In vitro release tests were studied using a USP Type-I apparatus at $37{\pm}0.5^{\circ}C$ and 100 rpm. The similarity factor (f2) was also used to check the difference statistically. The DSC results indicated that the crystallinity of ibuprofen was changed to an amorphous state after extrusion in both double and single melt extrusion. Double melt extrudate with ibuprofen showed the desired release in acidic media (pH 1.2) in the first two hours and basic (pH 6.8) during six hours. Double melt extrudate with glimepiride showed faster release in 60 min of over 80%, whereas the single extrudate with glimepiride showed retarded release due to the interaction with HPMC. The similarity factor(f2) value was 28.5, which demonstrates that there were different drug release behavior between the double and single extrusion. Consequently, the double melt extrudated formulation was robust and gave the desired drug release pattern.

Feeding Effects of Whole Crop Rice based TMR on Growth Performance and Carcass Characteristics of Hanwoo Steers (사료용 벼 위주 TMR 급여가 거세 한우의 생장 능력 및 도체 특성에 미치는 영향)

  • Kim, Jong Geun;Zhao, Guoqiang;Liu, Chang;Nan, Wei Sheng;Kim, Hak Jin;Kim, Kyoung Hoon;Ahn, Eok Geun;Min, Hyung-Gyu
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.2
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    • pp.97-104
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    • 2019
  • This experiment was conducted to investigate the effect of whole crop rice (WCR) based TMR on growth performance and carcass characteristics of Hanwoo steers. WCR "Yeongwoo"was harvested at yellow ripen stage and ensiled for 60 days. The crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), in vitro dry matter digestibility (IVDMD) and total digestible nutrient (TDN) content was 8.4 %, 28.0 %, 53.8 %, 72.4 % and 66.8 %, respectively. For silage quality, pH was 4.37 and lactic and butyric acid content were 2.84 and 0.04 % in DM. Sixteen Hanwoo steers (8-mon-old) were allocated into either a control (commercial TMR) and WCR-TMR (WCR-based TMR) group. The TMR were fed according to the feeding stage phase: growing (Initiate~14 month), early fattening (15 month~21 month) and late fattening (22 month~30 month). The body weight of control group increased (P<0.05) until early fattening stage, but late growing stage of WCR-TMR group was higher than that of control (P<0.05). Average daily gain (ADG) was significantly greater (P<0.05) in WCR-TMR group (total 0.78 kg/head) compared to control (total 0.66 kg/head) except for late fattening stage. The marketing weight and carcass weight were higher in WCR-TMR group (726 vs 765 kg; 417.8 vs 450.4 kg). The back fat thickness (11.75 vs 13.00 mm), Longissimus dorsi area (88.00 vs $89.88cm^2$) and yield index (65.87 vs 64.30) were not different between the two groups (P>0.05) and also no difference in meat yield grade (A : B : C = 2 : 4 : 2). Marbling score (4.00 vs 4.13), meat color (4.75 vs 4.75), fat color (3.13 vs 2.88), texture (1.25 vs 1.50) and maturity (2.00 vs 2.00) were not significant difference between the two groups and meat quality grade ($1^{{+}{+}}:1^+:1:2:3=0:2:4:2:0$) was also not different. In conclusion, TMR feeding based on WCR silage showed superiority in carcass yield and ADG compared to control TMR. It is considered that the use of WCR for feed is a necessary option for the substitution of the imported forages and the government's policy for rice production adjustment.

Comparison of the accuracy of intraoral scanner by three-dimensional analysis in single and 3-unit bridge abutment model: In vitro study (단일 수복물과 3본 고정성 수복물 지대치 모델에서 삼차원 분석을 통한 구강 스캐너의 정확도 비교)

  • Huang, Mei-Yang;Son, Keunbada;Lee, Wan-Sun;Lee, Kyu-Bok
    • The Journal of Korean Academy of Prosthodontics
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    • v.57 no.2
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    • pp.102-109
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    • 2019
  • Purpose: The purpose of this study was to evaluate the accuracy of three types of intraoral scanners and the accuracy of the single abutment and bridge abutment model. Materials and methods: In this study, a single abutment, and a bridge abutment with missing first molar was fabricated and set as the reference model. The reference model was scanned with an industrial three-dimensional scanner and set as reference scan data. The reference model was scanned five times using the three intraoral scanners (CS3600, CS3500, and EZIS PO). This was set as the evaluation scan data. In the three-dimensional analysis (Geomagic control X), the divided abutment region was selected and analyzed to verify the scan accuracy of the abutment. Statistical analysis was performed using SPSS software (${\alpha}=.05$). The accuracy of intraoral scanners was compared using the Kruskal-Wallis test and post-test was performed using the Pairwise test. The accuracy difference between the single abutment model and the bridge abutment model was analyzed by the Mann-Whitney U test. Results: The accuracy according to the intraoral scanner was significantly different (P < .05). The trueness of the single abutment model and the bridge abutment model showed a statistically significant difference and showed better trueness in the single abutment (P < .05). There was no significant difference in the precision (P = .616). Conclusion: As a result of comparing the accuracy of single and bridge abutments, the error of abutment scan increased with increasing scan area, and the accuracy of bridge abutment model was clinically acceptable in three types of intraoral scanners.

In Vitro Differentiation of Human Amniotic Membrane-derived Stem Cells into Hepatocyte-like Cells (양막 유래 줄기세포의 간세포로의 분화 유도)

  • Kook, Min-Ji;Park, Soo-Yeon;Kang, Hyun-Mi;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.10 no.1
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    • pp.63-73
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    • 2006
  • This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

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Anti-allergic activities of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (유산균 발효에 의한 율피(Castanea crenata inner shell) 열수추출물의 아토피 피부 질환에 관한 효과 연구)

  • Choi, Mi-Ok;Kim, Bae-Jin;Jo, Seung-Kyeung;Jung, Hee-Kyoung;Lee, Jin-Tae;Kim, Hak-Yoon;Kweon, Dae-Jun
    • Food Science and Preservation
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    • v.20 no.4
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    • pp.583-591
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    • 2013
  • Atopic dermatitis (AD) is a common chronic inflammatory disease associated with a cutaneous hypersensitivity reaction to an allergen. Although the incidence of AD is increasing these days, therapeutics has yet to be developed for its treatment. The aim of this study was conducted in order to compare and investigate the characteristic between the Castanea crenata inner shell extract (CS) and the Castanea crenata inner shell extract fermented by Lactobacillus bifermentans (FCS) for an anti-atopic medication. The total polyphenol and flavonoid contents were similar to CS and FCS. In the DPPH and superoxide anion radical scavenging, the CS and FCS had the potential for antioxidant activities. Both of them did not exhibit cytotoxicity to HS68 cells. The evaluation of the anti-inflammatory activity in Raw264.7 cells demonstrated that the FCS has inhibited the LPS-induced production of nitric oxide as compared to the CS. The anti-atopic dermatitis test was done through the induction of DNCB in AD hairless mice. The FCS has inhibited the development of the atopic dermatitis-like skin lesion by transdermal water loss, melanin and erythema of the skin as compared to the CS. Moreover, the pro-inflammatory cytokine IL-$1{\beta}$ and TNF-${\alpha}$ production in hairless mice were inhibited by the FCS treatment. It indicates that the fermentation of the Castanea crenata inner shell has the potential for the treatment of atopic dermatitis.