• Title/Summary/Keyword: in vitro cultivation

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In vitro Antimutagenic and Genotoxic Effects of Sophora Radix Extracts (고삼추출물의 in vitro 항돌연변이원성과 유전독성 연구)

  • Cho, Hyeon-Jo;Yoon, Hyunjoo;Park, Kyung-Hun;Lee, Je-Bong;Shim, Chang-Ki;Kim, Jin Hyo;Jeong, Mi Hye;Oh, Jin-Ah;Kim, Doo-Ho;Paik, Min-Kyoung
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.335-342
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    • 2013
  • Sophorae radix extract (SRE) has been registered as an environment-friendly organic material that is widely used in the cultivation of crops in Korea. Matrine, the active ingredient in SRE, was reported as a toxic substance in the nervous system in mice. However, no information is available on its toxic effects in other organisms. Therefore, antimutagenicity and two kinds of genotoxicity tests (bacterial reverse mutation and chromosome aberration test) of two samples of SRE were investigated in this study. Antimutagenicity test was experimented by using bacterial reverse mutation test. In the reverse mutation test, Salmonella Typhimurim TA98, TA1535 and TA1537 were used to evaluate the mutagenic potential of SRE. Bacterial reverse mutation test was also performed on positive and negative control groups in the presence of the metabolic activation system (with S-9 mix) and metabolic non-activation system (without S-9 mix). In the chromosome aberration test, Chinese hamster lung cells were exposed to SRE for 6 or 24 hours without S-9 mix, or for 6 hours with S-9 mix. Negative and positive control groups were experimented for chromosome aberration test. As a result, the number of mutated colonies induced by 4-NQO were reduced by SRE treatment in all strains, indicating that SRE may have antimutagenic effects. Reverse mutation was not shown at all concentrations of SRE, regardless of application of the metabolic activation system. In the chromosomal aberration test, one of the SRE sample gave a suspicious positive result at 250 ${\mu}g/ml$ in the presence of S-9 mix. For the more adequate evaluation of the genotoxic potential of SRE samples, other in vivo genotoxicity study is needed.

Aggregation of Human Eyelid Adipose-derived Stem Cells by Human Body Fluids

  • Song, Yeonhwa;Yun, Sujin;Yang, Hye Jin;Yoon, A Young;Kim, Haekwon
    • Development and Reproduction
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    • v.16 no.4
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    • pp.339-351
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    • 2012
  • Fetal bovine serum (FBS) is the most frequently used serum for the cultivation of mammalian cells. However, since animal-derived materials might not be appropriate due to safety issues, allogeneic human serum (HS) has been used to replace FBS, particularly for the culture of human cells. While there has been a debate about the advantages of HS, its precise effect on human adult stem cells have not been clarified. The present study aimed to investigate the effect of HS on the human eyelid adipose stem cells (HEACs) in vitro. When HEACs were cultivated in a medium containing 10% HS, many cells moved into several spots and aggregated there. The phenomenon was observed as early as 9 days following 10% HS treatment, and 12 days following 5% HS plus 5% FBS treatment. However, the aggregation was never observed when the same cells were cultivated with 10% FBS or bovine serum albumin. To examine whether cell density might affect the aggregation, cells were seeded with different densities on 12-well dish. Until the beginning of aggregation, cells seeded at low densities exhibited the longest culture period of 16 days whereas cells seeded at high densities showed the shortest period of 9 days to form aggregation. The number of cells was $15.1{\pm}0.2{\times}10^4$ as the least for the low density group, and $29.3{\pm}2.8{\times}10^4$ as the greatest for the high density group. When human cord blood serum or normal bovine serum was examined for the same effect on HEACs, interestingly, cord blood serum induced the aggregation of cells whereas bovine serum treatment has never induced. When cells were cultivated with 10% HS for 9 days, they were obtained and analyzed by RT-PCR. Compared to FBS-cultivated HEACs, HS-cultivated HEACs did not express VIM, and less expressed GATA4, PALLD. On the other hand, HS-cultivated HEACs expressed MAP2 more than FBS-cultivated HEACs. In conclusion, human adult stem cells could move and form aggregates by the treatment with human body fluids.

Pseudomonas azotoformans HC5 Effective in Antagonistic of Mushrooms Brown Blotch Disease Caused by Pseudomonas tolaasii (버섯 세균갈색무늬병균(Pseudomonas tolaasii)에 항균활성을 가지는 미생물 Pseudomonas azotoformans HC5)

  • Lee, Chan-Jung;Yoo, Young-Mi;Han, Ju-Yeon;Jhune, Chang-Sung;Cheong, Jong-Chun;Moon, Ji-Won;Gong, Won-Sik;Suh, Jang-Sun;Han, Hye-Su;Cha, Jae-Soon
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.219-224
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    • 2014
  • A gram-negative bacterium was isolated from spent substrate of Agaricus bisporus and showed marked antagonistic activity against Pseudomonas tolaasii. The bacterium was identified as Pseudomonas azotoformans by based on the cultural, biochemical and physiological characteristics, and 16S rRNA gene sequence. The isolated bacterium was saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for P. tolaasii cell was not sufficient for inhibition in vitro. Control efficacy of Pseudomonas azotoformans HC5 to brown blotch of P. tolaasii was 73, 78, and 71% on A. bisporus, Flammulina velutipes, and Pleurotus ostreatus, respectively. In the future, the suppressive bacterium may be useful for development of a biocontrol system.

Optimization of Culture Condition for Enhancing the Probiotics Functions (프로바이오틱스의 기능성 향상을 위한 배양법)

  • Chang, Bo Yoon;Han, Ji Hye;Cha, Bum-Suk;Ann, Sung-Ho;Kim, Sung Yeon
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.295-301
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    • 2015
  • The functions of probiotics, particularly Lactic acid bacteria, have been studied in a range of human diseases, including cancer, infectious diseases, gastrointestinal disorders, and allergies. Among the many benefits associated with the consumption of probiotics, modulation of immune activity has received the most attention. This study aimed at investigating the improved immune stimulatory and stability of L. plantarum when cultivated on modified basal media supplemented with the Undaria pinnatifida co-cultured with L. plantarum. An in vitro test showed that U. pinnatifida media cultured L. plantarum is strong enough to survive in the gastric juice (gastric and bile acid). Mouse macrophage-derived cell lines RAW 264.7 was used to measured immune stimulating activity of L. plantarum. When U. pinnatifida media cultured by L. plantarum was NO and $TNF-{\alpha}$ production is significantly increased compared to basal media cultured L. plantarum. These results show that U. pinnatifida could be applied for a component for cultivation of L. plantarum. This optimized U. pinnatifida medium can be used the improving of stability and immune function on production of probiotics.

Hair Revitalization Effects of Streptococcus sp. Strain Culture Medium Filtrate (HAIRCLETM) (Streptococcus sp. 배양 여과물(HAIRCLETM)의 모발 개선 효과)

  • Park, Hye Rim;Kim, Ha Yeon;Kim, Jin Young;Oh, Sinseok;Kang, Pilsun;Lee, Gang Hyuk;Lim, Jung-Hyurk;Shin, Song Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.2
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    • pp.169-180
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    • 2022
  • Streptococcus sp. is generally known as a strain that produces hyaluronic acid. In this study, we focused on ingredients other than hyaluronic acid among several metabolites produced by Streptococcus sp. during cultivation. The hair experimental sample (HAIRCLETM) was prepared with Streptococcus sp. culture filtrate under the condition that hyaluronic acid was not produced and the effect on the hair was identified. Tensile strength of hair was tested with a universal testing machine (UTM). Surface properties of a hair were examined with an atomic force microscope (AFM). Our results suggest that HAIRCLETM could prevent damage to a hair such as destroyed-cuticle, increased-faction force, and broken-hair. Furthermore, hair-related experiments confirmed that HAIRCLETM could promote the expression of hair growth factors VEGF, HGF, and Wnt10b and mitigate male hair loss by inhibiting androgen receptors and TGF-β2 expression. In addition, it was confirmed that the expression of barrier-related factors (INV, CLDN-1) was increased for oxidative stress, and the COX-2 expression, an inflammatory factor, was inhibited, thereby improving damaged scalp barriers and alleviating induced inflammation.

Selection of Salt-Tolerant Silage Rice Through in vitro Screening and Saltol QTL Analysis (기내 선발과 Saltol QTL 분석을 통한 내염성 증진 사료용 벼 선발)

  • Cho, Chuloh;Kim, Kyung Hwa;Ahn, Eok-Keun;Park, Hyangmi;Choi, Man-Soo;Chun, Jaebuhm;Seo, Mi-Suk;Jin, Mina;Kim, Dool-Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.214-221
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    • 2020
  • Salinity is one of the major abiotic stressors that inhibits the growth, yield, and productivity of crop plants. Therefore, it is necessary to develop crops with increased salt tolerance for cultivation in saline soils such as is found in reclaimed land. The objective of this study was to develop a salt-tolerant silage rice line that grows on reclaimed land. In order to develop this salt-tolerant silage rice, we transferred Saltol, a major QTL associated with salt tolerance, from IR64-Saltol, a salt-tolerant indica variety, into Mogyang, a susceptible elite japonica variety. To determine the effect of salt stress, Mogyang and IR64-Saltol cultivars were grown on a medium containing various concentrations of NaCl in in vitro conditions. Shoot length was found to decrease with increasing salt concentrations, and root growth was almost arrested at NaCl concentrations over 50 mM in the Mogyang cultivar. Based on these preliminary results, we screened five salt-tolerant lines showing superior growth under salt stress conditions. Polymerase chain reaction and sequencing results showed that the introgression types of Saltol QTL were derived from the IR64-Saltol cultivar in almost all selected lines. Based on the observed growth and physiological characteristics, the new Saltol introgression lines showed higher salt tolerance compared to the Mogyang parental cultivar. The salt-tolerant lines identified in this study could be used as a genetic resource to improve rice salt tolerance.

Evaluation of Characteristics, Winter Survival and Forage Production for Warm Season Grass in the Mid-Southern Regions of Korea (중남부지역에서 난지형 목초의 생육특성, 월동성 및 사초생산성 평가)

  • Park, Hyung Soo;Jung, Min Woong;Jung, Yong Bok;Lim, Young Chul;Choi, Ki Choon;Kim, Ji Hye;Lee, Ki Won;Choi, Gi Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.1
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    • pp.1-8
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    • 2014
  • Field studies were conducted from the years 2009 to 2012 in order to determine the cultivation limit as well as to evaluate the characteristics and forage production of warm season grass in Korea. Two bermudagrass [Cynodon dactylon (L.) Pers.] cultivars, two bahiagrass (Paspalum notatum Flugge) cultivars and a Kleingrass [Panicum coloratum L.] cultivar were compared for forage production and quality in the mid-southern regions of Korea. The experimental design was a randomized block design (RBD) with three replications. The number of days to seedling emergence for bremudagrass and bahiagrass was observed as approximately 12 days and 28 days after seeding, respectively. In Kwangju, the heading dates of bahiagrass and kleingrass were 21 August and 10 July, respectively,. Warm season grass did not winter in the mid-regions (Kimjea, Cheonan) of Korea. All of the Bermudagrass cultivars had higher dry matter (DM) than bahiagrass at the first harvest. The dry matter yield of kleingrass was usually greater than the other entries at all study sites. Peak forage DM production of bermudagrass and bahiagrass cultivars occurred in June and July, respectively. The contents of crude protein (CP) and total digestibility nutrient (TDN) for bermudagrass cultivars were usually greater than the other entries at all study sites. Further, acid detergent fiber (ADF) and in vitro DM digestibility (IVDMD) were similar across all cultivars.

Dual Coating Improves the Survival of Probiotic Bifidobacterium Strains during Exposure to Simulated Gastro-Intestinal Conditions (위장관내 조건에서 이중코팅 처리 된 프로바이오틱 비피도박테리움의 생존력 향상)

  • Kang, Joo Yeon;Lee, Do Kyung;Park, Jae Eun;Kim, Min Ji;Lee, Joong-Su;Seo, Jae-Gu;Chung, Myung Jun;Shin, Hea Soon;Ha, Nam Joo
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.275-281
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    • 2013
  • Probiotics have been reported to benefit human health by modulating immunity, lowering cholesterol, improving lactose tolerance, and preventing some cancer. Once ingested, probiotic microorganisms have to survive harsh conditions such as low pH, protease-rich condition, and bile salts during their passage through the gastro-intestinal (GI) tract colonize and proliferate to exert their probiotic effects. The dual coating technology, by which the bacteria are doubly coated with peptides and polysaccharides in consecutive order, was developed to protect the ingested bacteria from the harsh conditions. The aim of the study was to evaluate the viable stability of a doubly coated blend of four species of Bifidobacterium by comparing its bile/acid resistance and heat viability in vitro with that of the non-coated blend. After challenges with acid, bile salts, heat, and viable cell counts (VVCs) of the dual coated and non-coated blend were determined by cultivation on agar plates or flow cytometric measurement after being stain with the BacLigtht kit$^{TM}$. The results showed that the dual coated blend was much higher resistant to the acidic or bile salt condition than the non-coated blend and heat viability was also higher, indicating that the dual coating can improve the survival of probiotic bacteria during their transit through the GI tract after consumption.

Effect of tetracycline-HCl root conditioning on gingival epithelial cell attachment to root surface (염산테트라싸이클린으로 처리한 치근면의 치은상피세포부착에 관한 연구)

  • Hwang, Na-Young;Park, Byung-Ki;Kim, Sang-Mok;Kuk, Jung-Ki;Park, Ju-Chul;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.13-23
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    • 2002
  • The ultimate goal of periodontal therapy is directed to arresting the progression of the disease, and regenerating the fibrous attachment. In order to achieve such treatment aim, the plaque and calculus must be eliminated and the physiological conditions of the root surface must be changed to facilitate the attachment and migration of the new fibroblasts, The method of changing the proper root surface conditions to promote the healing of periodontal tissue involves mechanical procedures, such as scaling and root planing, and chemical procedures such as tetracycline-HCl. However, the formation of a long junctional epithelium was most frequently observed type of healing. Thus, the aim of this study was to examine in vitro the influence of surface conditioning of dentin by TC-HCl on human gingival epithelial cell attachment. Human gingival epithelial cells were obtained from healthy retromolar pad area(under the age 23 years). Seventy two teeth extracted from severe periodontitis were used as study material. To evaluate the epithelial cell attachment to dentin, the prepared specimen was divided to four groups. For the control group, only scaling and root planing were carried out, and for the test group, 1 to 3, the concentration of the TC-HCl was 50, 125 and 250mg/ml respectively. After cell cultivation time of 1-, 3-. 24 hour, for the indirect quantitative assessment of gingival epithelial cell attached to dentin sample, the absorbance of epithelial cell unattached to dentin was measured. The results were as follows; 1. There was no statistically significant difference between scaling and root planing group and TC-HCl 50mg/ml 125mg/ml and 250mg/ml group about absorbance of unattached epithelial cell to dentin sample(p>0.5). 2. As time passes, the absorbance of unattached gingival epithelial cell to dentin sample was decreased statistically significant(p<0.05). 3. There was no statistically significant difference among the TC-HCl group(p>0.05) We concluded that there was similar effect on gingival epithelial cell attachment between TC-HCl conditioning on root surface and only scaling and root planing treatment

Screening of Antifungal Activities of Plant Extracts against Phytopathogenic Fungi (식물추출물의 식물병원성 곰팡이 포자에 대한 발아억제 활성)

  • Park, Sang-jo;Rhu, Young Hyun;Bae, Soo Gon;Seo, Dong Hwan
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.343-351
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    • 2017
  • Plant extracts were screened for antifungal activity against major plant pathogens, Botrytis sp., Collectotrichum sp., Alternaria sp. and Cylindrocarpon sp. using 96-well microdilution method. Among the 662 methanol extracts from 401 plant species, 36 extracts showed complete inhibition of spore germination against at least one of four pathogenic fungi. Extracts of Morus alba twig and Sophora flavescens root showed minimum inhibition concentration (MIC) at $1,250{\mu}g/ml$ against Botrytis sp.. Extracts of Chloranthus japonicus root showed MIC at $1,250{\mu}g/ml$ against Collectotrichum sp.. Extracts of Glycyrrhiza uralensis aerial part, Inula helenium root and Menispermum dauricum root showed MIC between 625 and $1,250{\mu}g/ml$ against Alternaria sp.. G. uralensis aerial part and I. helenium root showed MIC at $1,250{\mu}g/ml$ against Cylindrocarpon sp.. Specifically, the extracts of Agrimonia pilosa root, Angelica tenuissima root, Asarum sieboldii root, Campsis grandifolia leaf and twig, Cnidium officinale root, Dictamnus dasycarpus root, G. uralensis aerial part, I. helenium root and M. alba twig completely inhibited spore germination at lower than $5,000{\mu}g/ml$ against all of four pathogenic fungi. Two methanol extracts from G. uralensis aerial part and M. alba twig may used as a candidate to develop into effective disease management materials in plant cultivation.