• Title/Summary/Keyword: cultured

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Induction of Apoptosis in HT-29 Human Colorectal Cancer by Aloin (인간 대장암 세포 HT-29에서 Aloin에 의한 Apoptosis 유도)

  • Yoo, Eun-Seon;Woo, Joong-Seok;Kim, Sung-Hyun;Lee, Jae-Han;Han, So-Hee;Jung, Soo-Hyun;Park, Young-Seok;Kim, Byeong-Soo;Kim, Sang-Ki;Park, Byung-Kwon;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.495-501
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    • 2019
  • Aloin [1,8-Dihydroxy-10-(${\beta}$-D-glucopyranosyl)-3-(hydroxymethyl)-9(10H)-anthracenone], is a natural anthraquinone from aloe. It has been shown to have antioxidant and anticancer effects in various types of human cancer cells, but the anticancer effects of aloin in human colorectal cancer cells HT-29 have not been elucidated. In this study, possible mechanisms by which aloin exerts its apoptotic action in cultured human colorectal cancer HT-29 cells were investigated. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay shows that treatment with aloin (0, 100, 200, 300 and $400{\mu}M$) reduced cell viability in a concentration-dependent manner in HT-29 and showed no effects on cell proliferation in A375SM and AGS cells. In addition, it was confirmed that apoptotic body was significantly increased as shown by 4',6-diamidino-2-phenylindole (DAPI) staining, and increased apoptosis rate by flow cytometry in HT-29 cells treated with aloin (0, 200 and $400{\mu}M$). We confirmed by western blotting that aloin activated Bax (pro-apoptotic), cleaved-poly (ADP-ribose) polymerase (PARP) and caspase-3, -8 and Bcl-2 (anti-apoptotic) were not changed compared with the control. Aloin induced up-regulation of phospho-p38 and down-regulation of phospho-extracellular signal-regulated kinase (ERK)1/2. Therefore, aloin suppressed the growth inhibitory effects by the induction of apoptosis in human colorectal cancer cells and has potential as a cancer preventive medicine.

Simultaneous Detection of Seven Phosphoproteins in a Single Lysate Sample during Oocyte Maturation Process (난자성숙 과정의 단일 시료에서 일곱 가지 인산화 단백질의 동시 분석 방법)

  • Yoon, Se-Jin;Kim, Yun-Sun;Kim, Kyeoung-Hwa;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.187-197
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    • 2009
  • Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

Effect of Different Levels of Applications of Illite on the Growth of Red Pepper (Capsicum annuum L.) in Bed Soil (상토에서 일라이트의 혼합비율에 따른 고추 육묘시 생육효과)

  • Lee, Seok-Eon;Kim, Hong-Ki;Kwon, Sang-Moon;Kim, Hee-Jung;Yoo, Ri-Bi;Baek, Ki-Tae;Lee, Moon-Soon;Woo, Sun-Hee;Park, Man;Chung, Keun-Yook
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.6
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    • pp.852-857
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    • 2010
  • This study was performed to explore the effect of the clay mineral illite on the improvement of bed soil and plant growth. Red pepper (Capsicum annuum L.) was used as a model vegetable crop. The experiment was performed during the whole six weeks in the glass house of the Chungbuk National University. Its seedlings were cultivated in the bed soil normally used for horticultural purpose. Of the seedlings cultured, the healthy and regular size of seed were selected and cultivated in the pots. They were treated with two forms of illite, particulate (PA) and powder (PW), at the following application rates: standard application[P1 (PA1, PW1), 1:20 (w/w)], two times[P2 (PA2, PW2), 1:10 (w/w)], and four times[P4 (PA4, PW4), 1:5 (w/w)] of standard application. Untreatment (P0) was used as a control pot. At six weeks of cultivation, their growth lengths were correspondingly increased as the application rate was increased ranging from P0, P1, P2, and to P4. Their growth length was a little greater on the application of powder illite (PW) than on the particulate illite (PA). Based on the plant analysis for the root, leaf, stem of red pepper, the uptake amounts of K, Ca, and Mg, were correspondingly increased, as the application rate was increased ranging from P0, P1, P2, and to P4, respectively. At the same application rate, their amounts taken up in the respective parts were higher on the application of PW illite than on the PA one. Especially the amounts of Ca and Mg were higher in the stem, leaf than root. Consequently, it appears that the illite treatment, especially, PW form of illite, enhance the growth of red pepper in the glass house during the whole six weeks of experiment.

Development of Vivorium, a new indoor horticultural ornamental plants via plant tissue culture techniques (식물조직배양 기술을 이용한 새로운 실내 원예 장식품인 비보리움(Vivorium)의 개발)

  • Hwang, Min Hee;Kim, Do Yeon;Cho, In Sun;Kim, Mi Hyung;Kwon, Hyun Sook;Kim, Jong Bo;Kim, Su Jung;Kim, Sun Hyung
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.179-185
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    • 2021
  • Indoor gardening includes wall greening, terrariums, and flower arrangements. Among these types of indoor gardens, the terrarium is easy to access for the general public, but in Korea, because of the focus on esthetics, the original purpose of creating terrariums, which was to grow plants sustainably in an enclosed space, has been lost. In addition, miniaturization of plants is required to grow plants in an enclosed space. Since the available plant species suitable for a terrarium are limited, only plants such as succulents, cacti, and moss have been used. In this study, Bronze (X Graptosedum) was used, and these problems were solved using the following three methods: placement and growth of virus-free plants in the terrarium; extending the diversity of plants with minimal size that can be planted in terrariums; and reducing the price of in vitro plants with minimal size by achieving large-scale production. In particular, tissue-cultured succulents were developed into a Vivorium by replacing the tissue culture container and renewing the composition of the plant. This paper suggests a new indoor horticultural field, Vivorium, that can improve the current limitations of terrariums and make them more accessible to the general public. The introduction and popularization of new indoor gardening fields with the increase in single-person households and indoor activities in the Pandemic era can also improve psychological stability among people and in the society.

Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV (시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이)

  • Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.387-401
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    • 2021
  • Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Assessing Effects of Calcium Chloride (CaCl2) Deicing Salt on Salt Tolerance of Miscanthus sinensis and Leachate Characterizations (염화칼슘 제설제 처리농도에 따른 참억새의 내염성 및 침출수 평가)

  • Ju, Jin-Hee;Yang, Ji;Park, Sun-Young;Yoon, Yong-Han
    • Journal of the Korean Institute of Landscape Architecture
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    • v.47 no.4
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    • pp.61-67
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    • 2019
  • The purpose of this research is to evaluate the salt tolerance of the Miscanthus sinensis and to characterize the content of pigments in the leachate in relation to calcium chloride ($CaCl_2$) deicing salt. Miscanthus sinensis were cultured at five different concentrations of calcium chloride deicing salt, 0, 1, 2, 5, and $10g{\cdot}L^{-1}$ (referred to Cont. C1, C2, C5, and C10) for four months. The salt tolerance and leachate while growing Miscanthus sinensis on soil which was artificially contaminated by calcium chloride deicing salt. Soil chemical properties (pH, E.C., $Ca^{2+}$, $Na^+$, $K^+$, and $Mg^{2+}$) and plant growth parameters (plant height, leaf length, leaf width, number of leaves, shoot fresh weight, root fresh weight, shoot dry weight, an root dry weight) were evaluated. Soil pH decreased, while electrical conductivity significantly decreased ($p{\leq}0.05$) with a higher concentration of deicing salt $0g{\cdot}L^{-1}$ (Cont.). The increase in the concentration of chloride-based exchangeable cations, along with the increase in the deicing salt treatments, were observed in $Ca^{2+}$ > $Na^+$ > $K^+$ > $Mg^{2+}$. Notably the $Ca^{2+}$ exchangeable cations were 83~90% higher than the others. The growth of Miscanthus sinensis significantly increased ($p{\leq}0.05$) with the concentration of deicing salt higher than $1g{\cdot}L^{-1}$ (C1) when compared to 0 g/L (Cont.), except for the $10g{\cdot}L^{-1}$ (C10) treatment. The results determined that the contamination of soil by deicing salt could negatively impact the soil and Miscanthus sinensis was a tolerant species for the deicing salts. Further research will be focused on soil improvement additives and the stable stimulated plant growth of Miscanthus sinensis and a formulation on that basis for the soil-plant continuum.

In vitro micropropagation of M.26 (Malus pumila Mill) apple rootstock and assessment of the genetic diversity of proliferated plantlets using simple sequence repeat markers (사과 대목 M.26 (Malus pumila Mill)의 기내 대량번식 및 simple sequence repeat 마커를 이용한 증식된 식물체의 유전적 다양성 평가)

  • Cho, Kang Hee;Han, Bong Hee;Han, Jeom Hwa;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan;Kim, Mi Young;Kim, Myung-Su
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.382-391
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    • 2018
  • The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

In vitro Multiplication of Hosta Tratt. Species Native to Korea by Shoot-tip Culture (경정배양에 의한 한국 자생 비비추속 식물의 기내증식)

  • Choi, Han;Yang, Jong Cheol;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Seung Youn
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.53-62
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    • 2019
  • The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

Mycelial and cultural characteristics of Pleurotus ostreatus 'Baekseon', a novel white cultivar for bottle culture (병재배용백색느타리신품종 『백선』의 균사배양및생육특성)

  • Choi, Jong In;Lee, Yun Hae;Gwon, Hee Min;Jeon, Dae Hoon;Lee, Yong Seon;Lee, Young Sun
    • Journal of Mushroom
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    • v.17 no.3
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    • pp.113-118
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    • 2019
  • Oyster mushrooms are an economically important crop, accounting for 35% of the total mushroom production in Korea. In this study, we developed a new cultivar of Pleurotus ostreatus, known as 'Baekseon,' which is characterized by a white pileus with a white stipe. It was bred by mating monokaryons isolated from white mutant oyster mushrooms that were naturally generated from 'Gonji-7ho' and 'Wonhyeong-1ho' at the Mushroom Research Institute, GARES, Korea in 2018. The optimum temperature for mycelial growth on potato dextrose agar medium was approximately $28-31^{\circ}C$, and the optimum temperatures for primordia formation and growth of fruit bodies on sawdust media were $22^{\circ}C$ and $20^{\circ}C$, respectively. The time required for the bottle-cultured mushrooms to complete spawn running, primordia formation, and growth of fruit bodies was 30 days, 4 days, and 4 days, respectively. The fruit bodies were bundle-shaped, the pilei were round type and white, and the stipes were white. The stipes were slender and longer than those of the control ('Miso'). In the productivity test, the yield per bottle was 185 g/1100 mL, which was 45% greater than that of the control ('Miso'). In the farm test, the yield per bottle for Farm A (Pyeongtaek) and Farm B (Yeoju) was 184 g/1100 mL and 178 g/850 mL, respectively. With regard to the physical properties of fruit bodies, the springiness, cohesiveness, gumminess, and brittleness of stipe tissue were 80%, 57%, 720 g, and 57 kg, respectively. These values were lower than those of the control ('Miso'). To test the shelf life, the fruit bodies were wrapped with antifogging film and stored at $4^{\circ}C$ for 28 days and then at room temperature for 4 days; such conditions were sufficient for maintaining edibility.