• Title/Summary/Keyword: subculture

Search Result 304, Processing Time 0.025 seconds

A Rapid Serotyping of Hydrophobic Strains of Mycobacterium acrofulaceum by Fluorescent Anti-Complement Technique (형광항보체법(螢光抗補體법)에 의한 Mycobacterium scrofulaceum 소수성균주(疏水性菌株)의 형별동정(型別同定))

  • Choi, Chul-Soon;Kim, Yong-Jae;Yang, Yong-Tae
    • The Journal of the Korean Society for Microbiology
    • /
    • v.15 no.1
    • /
    • pp.9-17
    • /
    • 1980
  • In the ecology and epidemiologic studies on various serotypes of atypical mycobacteria(AM), Schaefer's bacterial agglutination test(BA) provided the basis of the serologic procedures. Recently, attempts have been made to modify and to simplify the Schaefer's BA such as a slide agglutination test(Engel & Beerwald, 1970), a "simplified" BA(Reznikov & Leggo, 1972), an agglutination inhibition test(Richards & Eacret, 1972) and "micromethod"(Thoen et al., 1975). The BA, however, was not widely applied as a routine laboratory test mainly because it requires much times and labors to perform and partley because it is not applicable to hydrophobic strains either often encountered in the isolation of AM in the clinical bacteriology or stock strains maintained in the laboratory. On the contrary, fluorescent antibody technique with mycobacteria may have advantages over the BA because it is far more simpler in serologic procedures and is applicable to all strains of mycobacteria regardless of smooth or rough types of cultures. At the present, it is well known that the type-specific antigens are lacking on the surface of rough type of AM compared to that on smooth type of strain, but the antigenicity on the surface of the hydrophobic strains of AM which resulted from a series of subculture and the strain in the laboratory for 3 to 6 months has not been clarified. In this study, an attempt to serotype the hydrophobic strains of M. scrofulaceum serotype 41, 42 and 43 by fluorescent anti-complement(FAC) technique was made. The FAC technique with mycobacteria was also described in detail. In the summary, the complement fixing antibody titres of reference sera to smooth types of homologous serotype was highest, but the antibody titres of reference sera to hydrophobic strains of serotypes, 41, 42 and 43 gave two-to 8-folds lower than those to smooth type of strains. Although the sensitivity of type-specific antigens on the hydrophobic strains to reference sera was much lower, using the two units of reference sera determined by titration with hydrophobic strains, three serotypes, i. e., 41, 42 and 43 were specifically differentiated one another by FAC technique. This result indicated that the hydrophobic strains which were maintained in the laboratory at least for 6 months still retain type-specific antigen detectable by FAC technique.

  • PDF

Comparison of In Vitro Cell Transformation Assay Using Murine Fibroblasts and Human Keratinocytes

  • Ahn, Jun-Ho;Park, Sue-Nie;Yum, Yung-Na;Kim, Ji-Young;Lee, Michael
    • Toxicological Research
    • /
    • v.24 no.1
    • /
    • pp.37-44
    • /
    • 2008
  • The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

Plant Regeneration from Somatic Embryo with Structural Diversity from Leaf Explant Culture of Ostericum koreanum Kitagawa (강활(Ostericum koreanum Kitagawa)의 잎절편체 배양으로 부터 발생된 여러가지 형태의 체세포배를 통한 식물체 재생)

  • 조덕이;소웅영
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.1
    • /
    • pp.51-56
    • /
    • 1998
  • This study was carried out in order to establish plant regeneration via somatic embryogenesis from leaf explant of Ostericum koreanum Kitagawa and to elucidate the effects of NAA and cytokinins (kinetin, BA) on the abnormalities of somatic embryo and the relationship between thecotyledon numberand germinability. Calli were formed on leaf explants cultured on MS agar medium supplemented with various concentrations (0, 0.1, 0.5, 1, 2 mg/L) of NAA and cytokinins. The calli were white, watery and soft, became browning during cultures. Somatic embryos were formed from pale yellowish calli derived browning calli. High frequency somatic embryos were observed on MS medium containing 1 mg/L NAA and 0.1 mg/L BA after 60 days of culture. The mature somatic embryos germinated into plantlets without subculture after 2 weeks. The frequency of normal somatic embryo with two cotyledons was 39.8%. On the other hand, cotyledonary abnormalities of somatic embryos were observed at considerable frequency: 33.6% of somatic embryo with one cotyledon, 15.3% cotyledons with three, 8.2% four cotyledons and 3.1% jar shaped cotyledon. Germination frequency of somatic embryos with two cotyledons was 97.4%, and that of the embryos with abnormal cotyledon was almost similar to that of embryos with two cotyledons, except jar shaped somatic embryos (33.3%).

  • PDF

Micropropagation by Apical Meristem Culture of Wasabia japonica Matsum (고추냉이의 頂端分裂組織培養에 의한 微細增殖)

  • 은종선;고정애;김영선;김명준
    • Korean Journal of Plant Tissue Culture
    • /
    • v.24 no.1
    • /
    • pp.43-48
    • /
    • 1997
  • Apical meristems of Wasabia japonica were cultured on Murashige and Skoog's medium supplemented with cytokinins alone or together with 1.0 mg/L IAA. Shoot initials could be induced from leaf primordia on apical meristems. Calli and roots were formed on the medium containing cytokinins and 1.0 mg/L IAA in combination after 30 days of culture, but there were no callus proliferation. Shoot organogenesis began after 60 days of culture and these small shoots elongated when transferred to a medium containing 1.0 mg/L BA or kinetin. Shoots were formed directly without callus induction from apical meristems all the explants on the medium containing cytokinins variously, and most of the shoots proliferated multiple shoots which could be divided to obtain plantlets. Shoot multiplication rate in response to cytokinins was best on the medium containing 1.0 mg/L BA or 2.0 mg/L zeatin. Divided plantlets rooted well on MS medium containing 0.01 mg/L IBA after 15~30 days of subculture and the rooted plantlets developed into whole plants with multiple shoots. After rooting, the regenerated plants were washed and transferred to the pots containing sterilized soil.

  • PDF

Superoxide Dismutase Activity in Suspension Cultured Cells of Tomato (Lycopersicon esculentum Mill) (토마토(Lycopersicon esculentum Mill) 현탁배양세포에서 Superoxide Dismutase 활성)

  • 유순희;허경혜;권석윤;이행순;방재욱;곽상수
    • Korean Journal of Plant Tissue Culture
    • /
    • v.24 no.1
    • /
    • pp.57-61
    • /
    • 1997
  • We investigated changes in the superoxide dismutase (SOD) activity and SOD isoenzyme pattern in suspension cultures of tomato (Lycopersicon esculentum), which were compared with those of intact tomato plants. two grams (fr wt) of cells subcultured at 15-day intervals were inoculated into 50 mL MS medium containing l mg/L 2,4-D and 30 g/L sucrose in a 300 mL flask and maintained at $25^{\circ}C$ in the dark (100 rpm). The cell growth reached a maximum at 20 days after subculture (DAS), followed by a rapid decrease with further cultures. The cell colour changed from white to black from 23 DAS. The intracellular SOD activity (units/g cell dry wt) was significantly increased from 23 DAS and reached a maximum at 28 DAS (52,400 units), followed by a decrease with further cultures, whereas the extracellular SOD activity showed a maximum at 25 DAS (27,800 units/50 mL medium). The total SOD activity per flask showed a maximum at 25 DAS (35,700 units), in which the extracellular SOD activity occupied about 75%. The tomato cultured cells had four SOD isoenzymes and their patterns were well correlated with SOD activity without a qualitative change during the cell cultures. The intact tomato plants had an additional CuZnSOD isoenzyme, showing the different isoenzyme patterns from cultured cells.

  • PDF

Genotypic Effect of Watermelon (Citrullus lanatus Thunb.) on Organogenesis from Shoot Tip Culture of Seedlings (수박의 유묘 정단 배양시 유전적 차이가 기관 형성에 미치는 영향)

  • 이현기;백기엽;서영기;리왕영
    • Korean Journal of Plant Tissue Culture
    • /
    • v.21 no.4
    • /
    • pp.239-246
    • /
    • 1994
  • The genotypic (2n, 3n, 4n) response of watermelon in vitro shoot tip culture was evaluated. Different genotypes had similar response in terms of shoot formation and growth. Shoot formation was better at lower concentration of 0.3 mg/L BA and higher concentration of 5-10.0 mg/L 2iP and kinetin, but growth of newly formed shoot was inhibited. With further subculture, kinetin did not promote shoot formation Better shoot formation was observed at 0.3-0.5 mg/L BA. Combination of 0.3 mg/L BA and 0.3-0.5 mg/L BA was effective in shoot multiplication, growth and induction of more internodes. Varrying levels of light intensity and agar concentration did not affect the performance of tetraploid plants. Higher light intensity and agar concentrations decreased the number of shoot formed in triploid plane. Growth in both genotype, however was inhibited. Higher light intensity was found to promote leaf senescence in all genotypes. All growth inhibitors decreased the number of shoots formed and slowed plant growth there by prolonging duration of cultures. Growth inhibitors were to observed to decrease incidence of hyperhydricity in culture. No difference in shoot formation was observed in each of the concentrations used in Ancymidol, TIBA, CCC and PP333. Shoot formation and growth was more inhibited in ABA treatments. Leaf expansion and growth was poor in all treatments.

  • PDF

Somatic embryo induction and plant regeneration from cold-stored embryogenic callus of K. septemlobus (저온저장 음나무 배발생 캘러스로부터 체세포배 유도와 식물체 재생)

  • Lee, Na Nyum;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
    • /
    • v.42 no.4
    • /
    • pp.388-395
    • /
    • 2015
  • Somatic embryogenesis is as an excellent technology for potential use in plant mass production, germplasm conservation, or genetic engineering. We examined the effect of cold storage using 3 embryogenic callus lines with different levels of embryogenesis competence derived from immature zygotic embryo cultures of Kalopanax setemlobus. Somatic embryo induction, germination and plant conversion were evaluated after 1, 3 and 6 months storage at $4^{\circ}C$ in the dark. Most cold-stored embryogenic calli formed somatic embryos normally even after 6 months; however, the induction rate was gradually decreased by increasing the storage period. The most competent line tended to show a slight decline in somatic embryo induction rate, as compared with other lines after cold storage. In general, cold storage resulted in reduced somatic embryo germination and plant regeneration, although 93% somatic embryo germination and 91% plant conversion were achieved regardless of the storage period. Cold storage led to cell browning and degradation. Additionally, the cell structures were confirmed by the aceto-carmine and evans blue dye evaluation. Collectively, our results showed that embryogenic callus of K. septemlobus could be preserved at $4^{\circ}C$ without subculture for 6 months, and suggested the need for storage of relatively more competent embryogenic calli lines to support somatic embryo induction.

Ultrastructural Localization of Acid Phosphatase in Entamoeba histolytica and Entamoeba gingivalis (이질아메바 (Entamoeba histolytica)와 치은아메바 (Entamoeba gingivalis)의 Acid Phosphatase 활성에 관한 세포화학적 연구)

  • Cho, Kee-Mok;Cha, Hai-Young;Soh, Chin-Thack
    • Applied Microscopy
    • /
    • v.3 no.1
    • /
    • pp.1-16
    • /
    • 1973
  • A combined cytochemical and electron microscopic study was carried out for the demonstration of acid phosphatase activities in trophozoites of E. histolytica. and E. gingivalis. E. histolytica(YS-27) strain was isolated from liver abscess of 72-year-old man in September 1969, and E. gingivalis (YS-215) strain was collected from gingival crevice of 41-year-old man in January 1972. The amoeba strains were maintained by subculture on diphasic medium, and used throughout the study. The results are summarized as follows; 1. In E. histolytica, the reaction products were distributed evenly over the entire surface of plasma membrane, whereas E. gingivalis showed no activity of acid phosphatase on the plasma membrane, except in the portion of the uroid-like structure. 2. In the cytoplasm, various reaction precipitates were observed in vacuoles of both amoebae; vacuole limiting membrane, vacuole membrane and its contents and lysosome-like structure. Strong enzyme active contents but membrane reaction negative vacuoles were conspicuous in E. gingivalis. Endoplasmic reticulum showed a moderate activity. 3. Granule-like acid phosphatase reaction product was demonstrated in the nucleoplasm of E. gingivalis, but it was negative in E. histolytica.

  • PDF

Influence of Medium and Plant Growth Regulator on Micropropagation Efficiency in Blueberry (블루베리의 미세번식에서 배지와 식물생장조절제의 영향)

  • Kim, Hwa Young;Kang, Sun Pil;Hong, Sae Jin;Eum, Hyang Lan
    • Journal of Bio-Environment Control
    • /
    • v.24 no.3
    • /
    • pp.167-172
    • /
    • 2015
  • The aim of this study was to develop an effective production system of blueberry plants by using tissue culture technique. Murashige and skoog medium (MS) and woody plant medium (WPM) were compared for shoot formation of highbush blueberries. Also medium supplemented with zeatin/2-isopentenyl adenine (2iP)/benzyl aminopurine (BA) (1, 2/10, 15/4, $6mg{\cdot}L^{-1}$)and zeatin/2iP/BA (0.5/10, 15/$0.05mg{\cdot}L^{-1}$) as plant growth regulators to determine the effect of shoot formation and shoot proliferation, respectively. The shoot explants cultured on WPM showed higher shoot formation rates, more number of nodes, and longer root length than those on MS medium during the primary culture. Shoots were not formed when the explants were cultured on the medium without plant growth regulators or on only BA. The shoot explants cultured on the medium supplemented with 2iP showed low rates of shoot formation. On the other hand, zeatin was the most effective for shoot formation and growth of the explants. Also influence of different cytokinins (zeatin, 2iP) on the shoot proliferation of subcultured shoot explants was studied. There was no significant difference among the different concentrations of zeatin in the rate of shoot formation and number of shoots. However at higher concentration of zeatin, number of nodes was increased, and shoot length was shorted. The proper concentrations of zeatin for shoot propagation in subculture were found to be $0.5mg{\cdot}L^{-1}$ and $1mg{\cdot}L^{-1}$.

A Study on the Effects of Early Adolescents' Fandom Activities on Their Life Satisfaction: The Moderating Effect of Self-esteem (초기청소년의 팬덤활동이 삶만족도에 미치는 영향: 자아존중감의 조절효과)

  • Na, Jae-Eun
    • The Journal of the Korea Contents Association
    • /
    • v.17 no.8
    • /
    • pp.396-405
    • /
    • 2017
  • There have been constant studies on adolescents' fandom activities since the 2000s. Since their fandom activities are a subculture of adolescents, it is necessary to recognize it as such and to help them in a way that minimizes its adverse effects. In this regard, this study examined the effects of fandom activities on life satisfaction and examined whether or not self-esteem has a moderating effect. To this end, the study used the 6th year data of first graders of elementary school from Korea Youth Panel Survey conducted by Korea Youth Policy Institute and analyzed the data of a total of 1,088 of them. The results of this study are as follows: First, a negative correlation was found between their fandom activities and life satisfaction and between their fandom activities and self-esteem. A positive correlations was found between their fandom activities and life satisfaction, Second, it was found that adolescents' fandom activities negatively affected life satisfaction while self-esteem had a moderating effect on the relationship between fandom activities and life satisfaction. The significance of this study lies in the fact that it identified the variable that can moderate the negative role of adolescents' fandom activities, which provided a basis for future in-depth studies on the relationship between adolescents' fandom activities, individual internal variables, and life satisfaction.