• Title/Summary/Keyword: In vitro growth

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Fish Safety and Antimicrobial Activity of Natural Sulfur Solution on Aquatic Microorganisms (Saprolegnia parasitica) Isolated from Misgurnus mizolepis (미꾸라지(Misgurnus mizolepis)에서 분리된 수생균 (Saprolegnia parasitica)에 대한 천연유황수의 항균 활성 및 처리에 대한 어류 안전성)

  • Yi, Seung-Won;Lee, Seung-Hyeop;Lee, Sang-Jong;Kim, Mi-Hee;Lee, Hye-Hyun;Chu, Saet-Byul;Kim, Kyung-Hee;Lee, Hee Jung
    • Korean Journal of Environmental Biology
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    • v.35 no.2
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    • pp.116-122
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    • 2017
  • Basic dyes such as malachite green and methylene blue have been used as disinfectants to control water fungal infections since the 1930s. However, after succeeding reports of carcinogenicity and bioaccumulation of the dye, their use was forbidden in lieu of public health. This study undertook to evaluate the therapeutic effect of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica infection, and its safety in fish. In vitro antifungal evaluation of EM-PSS inhibited the growth of S. parasitica mycelia at concentrations of 50 ppm or higher. The acute toxicity test of EM-PSS to the mud fish (Misgurnus mizolepis) measured a no effect concentration (NOEC) at 100 ppm, the lowest effect concentration (LOEC) at 125 ppm, and the half-lethal concentration ($LC_{50}$) at 125 ppm in juvenile and 250 ppm in the immature stage. In addition, the ecotoxicity test of EM-PSS using Daphnia magna inhibited swimming of D. magna at concentrations of 100 ppm or less. Lastly, the EM-PSS prevented infection of S. parasitica to mud fish, at concentrations of 50 ppm. Furthermore, at 100 ppm concentration, the EM-PSS showed no acute toxicity on mud fish, nor any eco-toxic effects on D. magnano. Therefore, we conclude that carcinogenic disinfectants such as malachite green and methylene blue could be replaced by EM-PSS to remove S. parasitica in mud fish farming, and might be a potential eco-friendly disinfectant in aquaculture.

Biocompatibility and Histopathologic Change of the Acellular Xenogenic Pulmonary Valved Conduit Grafted in the Right Ventricular Outflow Tract (우심실 유출로에 이식한 무세포화 이종 폐동맥 판막도관의 생체 적합성 및 조직병리학적 변화양상에 대한 연구)

  • 허재학;김용진;박현정;김원곤
    • Journal of Chest Surgery
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    • v.37 no.6
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    • pp.482-491
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    • 2004
  • Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.

Effect of $Cd^{2+}$ on the Oocyte Maturation and Developmental Stages of Brown Frog Embryo, Rana dybowskii in vitro ($Cd^{2+}$이 북방산개구리의 난자성숙과 배아발달에 미치는 영향)

  • Ko Sun-Kun
    • Korean Journal of Environment and Ecology
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    • v.20 no.3
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    • pp.345-351
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    • 2006
  • This study investigates the toxic effects of $Cd^{2+}$on frog (Rana dybowskii) by the determination of oocyte maturation and development of embryo exposure to different concentrations of the toxicant. The results show that $Cd^{2+}$ concentration of 0.1ppm suppressed the maturation of the oocytes. To examine the reversibility of the inhibitory effects, the oocytes were exposed to the $Cd^{2+}$ only for 3 hours, and then transferred to plain medium and cultured further for 17 hours. The oocytes were recovered from the toxic effect of the $Cd^{2+}$ when they were exposed to 1ppm, but not to 2.5ppm of the $Cd^{2+}$. The development of 2 cell embryos to 32 cell was completely suppressed at 0.1ppm and the longer the embryos were exposed to the $Cd^{2+}$, the more damage appeared to the embryos and the cytolysis of the 32 cell was induced by $Cd^{2+}$ at 0.1ppm. On the other hand, the embryos of blastula stage were cultured 96 hours in presence of the $Cd^{2+}$ at various concentrations and were examined. The rates of mortality and malformed larvae were investigated by probit analysis. From the results of LC$_{50}$ of 0.1ppm and EC$_{50}$ of 0.08ppm, Tl of 5.0 was derived, which indicates $Cd^{2+}$ is to be considered a teratogenic compound. Such specific malformations occurred in 14.3% as spine deformations at the 0.05ppm, in 75.0% as tail deformations at the 0.1ppm, in 66.7% as abdominal deformations at the 0.01ppm and in 26.0% as profound deformations at the 0.1ppm of $Cd^{2+}$ concentration which living embryos were exposed to. $Cd^{2+}$ suppressed growth to head-tail length at 0.1ppm. In conclusion, The study results reveal that $Cd^{2+}$ must be considered highly toxic effect to oocyte maturation and embryonic development.

Anti-inflammation Activities of Cultured Products from Suspension Culture of Aloe vera Callus (Aloe vera Callus 현탁배양 생성물의 항염증 활성)

  • Kim, Myung Uk;Cho, Young Je;Lee, Shin Young
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.157-163
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    • 2013
  • Cultured products (callus and exopolysaccharide) were obtained from suspension culture of Aloe vera callus, and the extracts of callus were further prepared with cold water or 60% ethanol solution. The ethanol extract of callus (AC) and exopolysaccharide (ACP) of 10 mg/mL exhibited the relatively higher suppression activity of 43.2-52.1% against hyaluronidase activity. Thus, their anti-inflammatory effects were further investigated using animal cell (Raw 264.7) in vitro. Though AC shows a slight suppression effect of cell survival rate (97%) using MTT assay in the presence of $400{\mu}g/mL$ AC- dimethyl sulfoxide (DMSO), cell growth promotion was observed in the other samples of lower levels. It indicates that the ethanol extract of Aloe callus rarely affect cell survival rate in the ranges ($200-400{\mu}g/mL$) used in the study. Using Griess reagent, the suppression of NO production by the aloe callus extract was analyzed by measuring the amount of the nitrite produced in Raw 264.7 culture activated by lipopolysaccharide (LPS). As a result, supplementation of AC-distilled water (DW) and AC-DMSO produced higher levels of NO than the positive control LPS. However, the NO suppression effect by ACP-DW was so intense that lower amount ($80-100{\mu}g/mL$) suppressed NO production to the level of the control. The effect was attributed to the expression of the iNOS. Then, Raw 264.7 cells were stimulated with the LPS and expression of COX-2 protein level was analyzed depending on the Aloe suspension culture product treatment. The results showed that the ACP-DW supplemented medium did not express COX-2 by itself, and LPS stimulated COX-2 expression was slightly decreased. On the other hand, realtime-PCR analysis of the expression of inflammatory cytokine showed that IL-$1{\beta}$ and TNF-${\alpha}$ expression was highly suppressed in the ACP- distilled water supplemented medium.

Evaluation of time-dependent antimicrobial effect of sodium dichloroisocyanurate (NaDCC) on Enterococcus faecalis in the root canal (이염화이소시아뉼산나트륨 제재의 근관 내 사용 시 시간에 따른 E. faecalis에 대한 항균성 평가)

  • Kim, Hye-Jeong;Park, Se-Hee;Cho, Kyung-Mo;Kim, Jin-Woo
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.121-129
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    • 2007
  • The purpose of this study was to assess the antibacterial effect of sodium dichloroisocyanurate (NaDCC), sodium hypochlorite (NaOCl), and chlorhexidine (CHX) on Enterococcus faecalis and to evaluate and to compare the time-dependant antimicrobial effect of NaDCC with NaOCl and CHX in the root canal in vitro before and after instrumentation. Extracted Human single teeth were prepared by serial instrumentation technique. The samples were autoclaved and contaminated for 3 days with E. faecalis monocultures. The teeth were then divided into 4 groups Each group was irrigated and inserted with 2% NaOCl, 2% NaDCC, 2% CHX and steri)ized saline. After 6, 12, 24, 72h, and 1 week incubation, sterilized paper point was inserted into the root canal. Paper points containing root canal contents were then placed on the agar plate. And then each root cana) was prepared with #4 and #5 GG (Gates-Glidden) drill. The debris were collected in the sterilized microtube and the plates were incubated at $37^{\circ}C$ in an increased $CO_2$ atmosphere. After 24h incubation the growth of bacteria around the paper points were measured. NaOCl and NaDCC solution shows similar antimicrobial effect for E. faecalis at 6, 12, 24, 72h and 1 week. In centrol group, irrigated with sterilized saline, no antimicrobial effect was observed. The results are in agreement with other investigators, who have shown the bactericidal property and possibility of NaDCC as a root canal irrigation solution. Thus it seems that NaDCC solutions can be clinically applied into the root canal within 1 week after dilution.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Isolation and Identification of Competitive Fungi on Medium for Black Wood Ear Mushroom in Korea and In Vitro Selection of Potential Biocontrol Agents (목이버섯 배지 오염 곰팡이균의 분리, 동정 및 생물학적 방제제 선발)

  • Seoyeon Kim;Miju Jo;Sunmin An;Jiyoon Park;Jiwon Park;Sungkook Hong;Jiwoo Kim;Juhoon Cha;Yujin Roh;Da Som Kim;Mi jin Jeon;Won-Jae Chi;Sook-Young Park
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.66-77
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    • 2024
  • Black wood ear mushroom (Auricularia auricula-judae) is one of the most economically important mushrooms in China, Japan, and Korea. The cultivation of wood ear mushrooms on artificial substrates is more efficient in terms of time and cost compared with their natural growth on trees. However, if the substrate cultivation is infected by fast-growing fungi, the relatively slow-growing ear mushroom will be outcompeted, leading to economic losses. In this study, we investigated the competitive fungal isolates from substrates infected with fast-growing fungi for the cultivation of ear mushrooms in Jangheung and Sunchon, Korea. We collected 54 isolates and identified them by sequencing their internal transcribed spacer region with morphological identification. Among the isolates, the dominant isolates were Trichoderma spp. (92.6%), Penicillium spp. (5.6%), and Talaromyces sp. (1.8%). To find an appropriate eco-friendly biocontrol agent, we used five Streptomyces spp. and Benomyl, as controls against Trichoderma spp. and Penicillium spp. Among the six Streptomyces spp., Streptomyces sp. JC203-3 effectively controlled the fungi Trichoderma spp. and Penicillium spp., which pose a significant problem for the substrates of black wood ear mushrooms. This result indicated that this Streptomyces sp. JC203-3 can be used as biocontrol agents to protect against Trichoderma and Penicillium spp.

In Vitro Intrinsic Radiosensitivity Of Human Squamous Cell Carcinoma in Primary Culture (인체 상피 세포암의 일차 배양을 이용한 방사선 민감도 측정)

  • Choi Eun Kyung;Yang Kwang Mo;Yi Byong Yong;Chang Hyesook;Kim Sang-Yoon;Nam Joo-Hyun;Yu Eunsil;Lee Inchul
    • Radiation Oncology Journal
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    • v.12 no.1
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    • pp.27-31
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    • 1994
  • There are a number of reports suggesting that there may be a correlation between the clinical response to radiotherapy in various tumors and the clonogenic survival of cell lines derived from these tumors following exposure to 2 Gy(SF2). Authors conducted this study to determine SF2 for cells in primary culture from surgical specimens. The tumor tissues with squamous cell carcinoma of uterine cervix and head and neck were obtained. The tumor tissues were disaggregated to single cells by incubating with collagenase type w for 2 hours with constant stirring. Single cell suspensions were inoculated in four 24-well plates precoated with cell adhesive matrix. After 24 hours of incubation at 37$ ^{\circ}C $, rows of four wells were then irradiated, consisting of control set and five other sets each receiving doses of 1,2,3,4, and 6 Gy. After incubation for a total of 13 days, the cultures were stained with crystal violet and survival at each dose was determined by quantitative image analysis system, To determine whether cell growth was of epithelial origin, immunocytochemical staining with a mixture of cytokeratin and epithelial monoclonal antibodies were performed on cell cultures. During the period of this study, we received 5 squamous cell carcinoma specimens of head and neck and 20 of uterine cervical carcinoma. Of these, 15 yielded enough cells for radiosensitivity testing. This resulted an overall success rate of 60$ \% $. The mean SF2 value for 15 tumours was 0.55$\pm$0.17 ranging from 0.20 to 0.79. These results indicate that there is a broad range of sensitivities to radiation in same histologic type. So with a large patient population, we plan to determine whether a different SF2 value is associated with tumours that are controlled with radiotherapy than those that are not.

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Antitumor Activity of CBT-AK5 Purified from Lactobacillus casei against Sarcoma-180 Infected ICR Mice (유산균의 배양물로부터 분리된 물질에 대한 암 유발 ICR Mice의 항암 효과)

  • Yeo, Moon-Hwan;Kim, Dong-Myung;Kim, Yong-Hwan;Kim, Jung-Hwa;Baek, Hong;Chung, Myung-June
    • Journal of Dairy Science and Biotechnology
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    • v.26 no.2
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    • pp.23-30
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    • 2008
  • This study implicated that the CBT-AK5 purified from Lactobacillus casei (LAFTI L26) which showed antitumor activity in ICR mice. Hence, ICR mice were inoculated intraperitoneally Sarcoma-180 as well as CBT-AK5. Then we observed the life span and tumor increment of those ICR mice. Here our studies showed effect on two different way of treatment as intraperitoneally and orally treated in Sarcoma-180 infected ICR mice. We found that intraperitoneally treatment of Sarcoma-180 and CBT-AK5 is more effective than orally fed. The life span of the ICR mice were highly reduced after the inoculation of Sarcoma-180. Those effects like increment of body weight, the growth of ascites and solid were inhibited significantly after the treatment of CBT-AK5 in Sarcoma-180 infected ICR mice. Finally these studies suggested that CBT-AK5 isolated form Lactobacillus casei showed excellent antitumor activity against Sarcoma-180 infected ICR mice.

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The Expression of IL-8 and GRO$\alpha$/MGSA in HUVEC Stimulated by the TNF-$\alpha$ and IL-1 (TNF-$\alpha$와 IL-1 자극에 의한 제대정맥내피세포에서의 IL-8 및 GRO/MGSA의 발현)

  • Song, Jeong-Sup;Shin, Moon-Sun;Ahn, Joong-Hyun;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.3
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    • pp.338-349
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    • 1999
  • Polymorphonuclear leukocytes(PMN) are the predominant inflammatory cells recruited in acute lung injury such as adult respiratory distress syndrome, pneumonia and also chronic lung disease such as idiopathic pulmonary fibrosis and pulmonary emphysema. Interleukin-8(IL-8) is an 8,000 D protein produced by many cells and has potent neutrophil chemoattractant and activating properties. The GRO, also called melanoma growth-stimulatory activity(MGSA), referring to a peptide of 73 amino acids, was reported to be mitogenic for cultured human melanoma cells. Mature GRO/MGSA has marked sequence similarity to IL-8. In view of the structural similarities to IL-8, it was of particular interest to test GRO for neutrophil activating and chemotactic properties. We found a significant release of IL-8 and GRO/MGSA from the cultured human umbilical vein endothelial cell(HUVEC) which was stimulated either with TNF$\alpha$ or IL-1$\beta$ and also found the expression of IL-8 and GRO/MGSA mRNA. Neutrophil chemotactic activity was enhanced in accordance with the increased IL-8 and GRO/MGSA. Our study also suggest that the IL-8 is more important in the increased neutrophil chemotactic activity than GRO/MGSA when endothelial cell is stimulated with TNF$\alpha$ or IL-1$\beta$ in vitro.

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