WorldWideScience

Sample records for solid culture medium

  1. Growth evaluation of Lentinula edodes in solid medium cultures for mycelium production as inoculum

    OpenAIRE

    Villegas E Valeska; Pérez Ana Milena; Arredondo Clara

    2007-01-01

    Shitake (Lentinula edodes) Pegler jumbo strain growth was evaluated in different solid mediums and growth substrates for spawn production. Mycelium growth was tested in three culture mediums (MYA, OMYA, PDYA) at two pHs (5, 5.5), using two eucalyptus sawdust percentages (0.3%, 0.2%). Analysing variance revealed significant differences in culture medium (P0.05). The liquid inoculation technique was used for evaluating mushroom spawn production using five different combinations of eucalyptus sa...

  2. Growth evaluation of Lentinula edodes in solid medium cultures for mycelium production as inoculum

    Directory of Open Access Journals (Sweden)

    Valeska Villegas E

    2007-07-01

    Full Text Available Shitake (Lentinula edodes Pegler jumbo strain growth was evaluated in different solid mediums and growth substrates for spawn production. Mycelium growth was tested in three culture mediums (MYA, OMYA, PDYA at two pHs (5, 5.5, using two eucalyptus sawdust percentages (0.3%, 0.2%. Analysing variance revealed significant differences in culture medium (P0.05. The liquid inoculation technique was used for evaluating mushroom spawn production using five different combinations of eucalyptus sawdust and wheat grain, finding significant differences between treatments, the best combination for shiitake growth being 80% wheat grain and 20% eucalyptus sawdust.

  3. Influence of Dy in solid solution on the degradation behavior of binary Mg-Dy alloys in cell culture medium.

    Science.gov (United States)

    Yang, Lei; Ma, Liangong; Huang, Yuanding; Feyerabend, Frank; Blawert, Carsten; Höche, Daniel; Willumeit-Römer, Regine; Zhang, Erlin; Kainer, Karl Ulrich; Hort, Norbert

    2017-06-01

    Rare earth element Dy is one of the promising alloying elements for magnesium alloy as biodegradable implants. To understand the effect of Dy in solid solution on the degradation of Mg-Dy alloys in simulated physiological conditions, the present work studied the microstructure and degradation behavior of Mg-Dy alloys in cell culture medium. It is found the corrosion resistance enhances with the increase of Dy content in solid solution in Mg. This can be attributed to the formation of a relatively more corrosion resistant Dy-enriched film which decreases the anodic dissolution of Mg. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Bioleaching of fly ash from municipal solid waste incineration using kitchen waste saccharified solution as culture medium

    International Nuclear Information System (INIS)

    Wei, S.; Juan, W.; Qunhui, W.

    2013-01-01

    Summary: Reduced sugar in saccharified solution from kitchen waste was used as the carbon source. Domesticated A. niger AS 3.879C , which can withstand 20% of kitchen waste, was used as the inoculum in the bioleaching process of municipal solid waste incineration fly ash. The effect of reduced sugar concentration, fly ash concentration, and medium volume on the heavy metal extraction and yield of fly ash as well as the optimum bioleaching conditions; the inoculation amount of AS 3 .879C 1% (v/v), reduced sugar concentration of 80 g/l, fly ash concentration of 20 g/l, medium volume of 200 ml, and the addition of fly ash (20 g/l) after culturing for 4 days at 30 degree C and 140 r/min were obtained. Under the optimum condition, the extraction yield of the seven tested heavy metals are in the order of Cd > Zn > Cu > Mn > Pb > Cr > Fe; the extraction yield of Cd and Zn reached 88.7% and 73.1% respectively. Fly ash satisfied the Standard for Pollution Control on the Security Landfill Site for Hazardous Wastes (GB 18598-2001) after heavy metal extraction. (author)

  5. Direct blood culturing on solid medium outperforms an automated continuously monitored broth-based blood culture system in terms of time to identification and susceptibility testing

    Directory of Open Access Journals (Sweden)

    E.A. Idelevich

    2016-03-01

    Full Text Available Pathogen identification and antimicrobial susceptibility testing (AST should be available as soon as possible for patients with bloodstream infections. We investigated whether a lysis-centrifugation (LC blood culture (BC method, combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS identification and Vitek 2 AST, provides a time advantage in comparison with the currently used automated broth-based BC system. Seven bacterial reference strains were added each to 10 mL human blood in final concentrations of 100, 10 and 1 CFU/mL. Inoculated blood was added to the Isolator 10 tube and centrifuged at 3000 g for 30 min, then 1.5 mL sediment was distributed onto five 150-mm agar plates. Growth was observed hourly and microcolonies were subjected to MALDI-TOF MS and Vitek 2 as soon as possible. For comparison, seeded blood was introduced into an aerobic BC bottle and incubated in the BACTEC 9240 automated BC system. For all species/concentration combinations except one, successful identification and Vitek 2 inoculation were achieved even before growth detection by BACTEC. The fastest identification and inoculation for AST were achieved with Escherichia coli in concentrations of 100 CFU/mL and 10 CFU/mL (after 7 h each, while BACTEC flagged respective samples positive after 9.5 h and 10 h. Use of the LC-BC method allows skipping of incubation in automated BC systems and, used in combination with rapid diagnostics from microcolonies, provides a considerable advantage in time to result. This suggests that the usefulness of direct BC on solid medium should be re-evaluated in the era of rapid microbiology.

  6. Selective medium for culture of Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Cook, Beth S; Beddow, Jessica G; Manso-Silván, Lucía; Maglennon, Gareth A; Rycroft, Andrew N

    2016-11-15

    The fastidious porcine respiratory pathogen Mycoplasma hyopneumoniae has proven difficult to culture since it was first isolated in 1965. A reliable solid medium has been particularly challenging. Moreover, clinical and pathological samples often contain the fast-growing M. hyorhinis which contaminates and overgrows M. hyopneumoniae in primary culture. The aim of this study was to optimise the culture medium for recovery of M. hyopneumoniae and to devise a medium for selection of M. hyopneumoniae from clinical samples also containing M. hyorhinis. The solid medium devised by Niels Friis was improved by use of Purified agar and incorporation of DEAE-dextran. Addition of glucose or neutralization of acidity in liquid medium with NaOH did not improve the final yield of viable organisms or alter the timing of peak viability. Analysis of the relative susceptibility of M. hyopneumoniae and M. hyorhinis strains to four antimicrobials showed that M. hyopneumoniae is less susceptible than M. hyorhinis to kanamycin. This was consistent in all UK and Danish strains tested. A concentration of 2μg/ml of kanamycin selectively inhibited the growth of all M. hyorhinis tested, while M. hyopneumoniae was able to grow. This forms the basis of an effective selective culture medium for M. hyopneumoniae. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  7. Enhancement of fructosyltransferase and fructooligosaccharides production by A. oryzae DIA-MF in Solid-State Fermentation using aguamiel as culture medium.

    Science.gov (United States)

    Muñiz-Márquez, Diana B; Contreras, Juan C; Rodríguez, Raúl; Mussatto, Solange I; Teixeira, José A; Aguilar, Cristóbal N

    2016-08-01

    The aim of this work was to improve the production of fructosyltransferase (FTase) by Solid-State Fermentation (SSF) using aguamiel (agave sap) as culture medium and Aspergillus oryzae DIA-MF as producer strain. SSF was carried out evaluating the following parameters: inoculum rate, incubation temperature, initial pH and packing density to determine the most significant factors through Box-Hunter and Hunter design. The significant factors were then further optimized using a Box-Behnken design and response surface methodology. The maximum FTase activity (1347U/L) was obtained at 32°C, using packing density of 0.7g/cm(3). Inoculum rate and initial pH had no significant influence on the response. FOS synthesis applying the enzyme produced by A. oryzae DIA-MF was also studied using aguamiel as substrate. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Rapid Identification of Microorganisms from Positive Blood Culture by MALDI-TOF MS After Short-Term Incubation on Solid Medium.

    Science.gov (United States)

    Curtoni, Antonio; Cipriani, Raffaella; Marra, Elisa Simona; Barbui, Anna Maria; Cavallo, Rossana; Costa, Cristina

    2017-01-01

    Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a useful tool for rapid identification of microorganisms. Unfortunately, its direct application to positive blood culture is still lacking standardized procedures. In this study, we evaluated an easy- and rapid-to-perform protocol for MALDI-TOF MS direct identification of microorganisms from positive blood culture after a short-term incubation on solid medium. This protocol was used to evaluate direct identification of microorganisms from 162 positive monomicrobial blood cultures; at different incubation times (3, 5, 24 h), MALDI-TOF MS assay was performed from the growing microorganism patina. Overall, MALDI-TOF MS concordance with conventional methods at species level was 60.5, 80.2, and 93.8% at 3, 5, and 24 h, respectively. Considering only bacteria, the identification performances at species level were 64.1, 85.0, and 94.1% at 3, 5, and 24 h, respectively. This protocol applied to a commercially available MS typing system may represent, a fast and powerful diagnostic tool for pathogen direct identification and for a promptly and pathogen-driven antimicrobial therapy in selected cases.

  9. Rapid identification of microorganisms from positive blood cultures by MALDI-TOF mass spectrometry subsequent to very short-term incubation on solid medium.

    Science.gov (United States)

    Idelevich, E A; Schüle, I; Grünastel, B; Wüllenweber, J; Peters, G; Becker, K

    2014-10-01

    Rapid identification of the causative microorganism is important for appropriate antimicrobial therapy of bloodstream infections. Bacteria from positive blood culture (BC) bottles are not readily available for identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Lysis and centrifugation procedures suggested for direct MALDI-TOF MS from positive BCs without previous culture are associated with additional hands-on processing time and costs. Here, we describe an alternative approach applying MALDI-TOF MS from bacterial cultures incubated very briefly on solid medium. After plating of positive BC broth on Columbia blood agar (n = 165), MALDI-TOF MS was performed after 1.5, 2, 3, 4, 5, 6, 7, 8, 12 and (for control) 24 h of incubation until reliable identification to the species level was achieved (score ≥2.0). Mean incubation time needed to achieve species-level identification was 5.9 and 2.0 h for Gram-positive aerobic cocci (GPC, n = 86) and Gram-negative aerobic rods (GNR, n = 42), respectively. Short agar cultures with incubation times ≤2, ≤4, ≤6, ≤8 and ≤12 h yielded species identification in 1.2%, 18.6%, 64.0%, 96.5%, 98.8% of GPC, and in 76.2%, 95.2%, 97.6%, 97.6%, 97.6% of GNR, respectively. Control species identification at 24 h was achieved in 100% of GPC and 97.6% of GNR. Ethanol/formic acid protein extraction performed for an additional 34 GPC isolates cultivated from positive BCs showed further reduction in time to species identification (3.1 h). MALDI-TOF MS using biomass subsequent to very short-term incubation on solid medium allows very early and reliable bacterial identification from positive BCs without additional time and cost expenditure. © 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.

  10. Rapid Detection and Identification of Candidemia by Direct Blood Culturing on Solid Medium by Use of Lysis-Centrifugation Method Combined with Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

    Science.gov (United States)

    Idelevich, Evgeny A.; Grünastel, Barbara

    2016-01-01

    ABSTRACT Candida sepsis is a life-threatening condition with increasing prevalence. In this study, direct blood culturing on solid medium using a lysis-centrifugation procedure enabled successful Candida species identification by matrix-assisted laser desorption–ionization time of flight mass spectrometry on average 3.8 h (Sabouraud agar) or 7.4 h (chocolate agar) before the positivity signal for control samples in Bactec mycosis-IC/F or Bactec Plus aerobic/F bottles, respectively. Direct culturing on solid medium accelerated candidemia diagnostics compared to that with automated broth-based systems. PMID:27795344

  11. Rapid Detection and Identification of Candidemia by Direct Blood Culturing on Solid Medium by Use of Lysis-Centrifugation Method Combined with Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Idelevich, Evgeny A; Grünastel, Barbara; Becker, Karsten

    2017-01-01

    Candida sepsis is a life-threatening condition with increasing prevalence. In this study, direct blood culturing on solid medium using a lysis-centrifugation procedure enabled successful Candida species identification by matrix-assisted laser desorption-ionization time of flight mass spectrometry on average 3.8 h (Sabouraud agar) or 7.4 h (chocolate agar) before the positivity signal for control samples in Bactec mycosis-IC/F or Bactec Plus aerobic/F bottles, respectively. Direct culturing on solid medium accelerated candidemia diagnostics compared to that with automated broth-based systems. Copyright © 2016 American Society for Microbiology.

  12. Medium-energy ion reflection from solids

    CERN Document Server

    Mashkova, ES

    1985-01-01

    ``Medium-Energy Ion Reflection from Solids'' analyses the results of experimental, theoretical and computer investigations on the process of scattering of ions by solid surfaces. Surface scattering is a relatively young and rapidly developing branch of the physics of atomic collisions and the literature on this subject has rapidly grown.As the first monograph devoted specifically to surface scattering of ions, this book is directed at scientists involved in ion-solid interaction studies.

  13. MALDI-TOF mass spectrometry following short incubation on a solid medium is a valuable tool for rapid pathogen identification from positive blood cultures.

    Science.gov (United States)

    Kohlmann, Rebekka; Hoffmann, Alexander; Geis, Gabriele; Gatermann, Sören

    2015-01-01

    Rapid identification of the causative microorganism is a key element in appropriate antimicrobial therapy of bloodstream infections. Whereas traditional analysis of positive blood cultures requires subculture over at least 16-24h prior to pathogen identification by, e.g. matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), sample preparation procedures enabling direct MALDI-TOF MS, i.e. without preceding subculture, are associated with additional effort and costs. Hence, we integrated an alternative MALDI-TOF MS approach in diagnostic routine using a short incubation on a solid medium. Positive blood cultures were routinely plated on chocolate agar plates and incubated for 4h (37 °C, 5% CO2). Subsequently, MALDI-TOF MS using a Microflex LT instrument (Bruker Daltonics) and direct smear method was performed once per sample. For successful identification of bacteria at species level, score cut-off values were used as proposed by the manufacturer (≥ 2.0) and in a modified form (≥ 1.5 for MALDI-TOF MS results referring to Gram-positive cocci and ≥ 1.7 for MALDI-TOF MS results referring to bacteria other than Gram-positive cocci). Further data analysis also included an assessment of the clinical impact of the MALDI-TOF MS result. Applying the modified score cut-off values, our approach led to an overall correct species identification in 69.5% with misidentification in 3.4% (original cut-offs: 49.2% and 1.8%, respectively); for Gram-positive cocci, correct identification in 68.4% (100% for Staphylococcus aureus and enterococci, 80% for beta-hemolytic streptococci), for Gram-negative bacteria, correct identification in 97.6%. In polymicrobial blood cultures, in 72.7% one of the pathogens was correctly identified. Results were not reliable for Gram-positive rods and yeasts. The approach was easy to implement in diagnostic routine. In cases with available clinical data and successful pathogen identification, in 51.1% our

  14. Solid H2 in the interstellar medium

    Science.gov (United States)

    Füglistaler, A.; Pfenniger, D.

    2018-06-01

    Context. Condensation of H2 in the interstellar medium (ISM) has long been seen as a possibility, either by deposition on dust grains or thanks to a phase transition combined with self-gravity. H2 condensation might explain the observed low efficiency of star formation and might help to hide baryons in spiral galaxies. Aims: Our aim is to quantify the solid fraction of H2 in the ISM due to a phase transition including self-gravity for different densities and temperatures in order to use the results in more complex simulations of the ISM as subgrid physics. Methods: We used molecular dynamics simulations of fluids at different temperatures and densities to study the formation of solids. Once the simulations reached a steady state, we calculated the solid mass fraction, energy increase, and timescales. By determining the power laws measured over several orders of magnitude, we extrapolated to lower densities the higher density fluids that can be simulated with current computers. Results: The solid fraction and energy increase of fluids in a phase transition are above 0.1 and do not follow a power law. Fluids out of a phase transition are still forming a small amount of solids due to chance encounters of molecules. The solid mass fraction and energy increase of these fluids are linearly dependent on density and can easily be extrapolated. The timescale is below one second, the condensation can be considered instantaneous. Conclusions: The presence of solid H2 grains has important dynamic implications on the ISM as they may be the building blocks for larger solid bodies when gravity is included. We provide the solid mass fraction, energy increase, and timescales for high density fluids and extrapolation laws for lower densities.

  15. CHROMagar Orientation Medium Reduces Urine Culture Workload

    Science.gov (United States)

    Manickam, Kanchana; Karlowsky, James A.; Adam, Heather; Lagacé-Wiens, Philippe R. S.; Rendina, Assunta; Pang, Paulette; Murray, Brenda-Lee

    2013-01-01

    Microbiology laboratories continually strive to streamline and improve their urine culture algorithms because of the high volumes of urine specimens they receive and the modest numbers of those specimens that are ultimately considered clinically significant. In the current study, we quantitatively measured the impact of the introduction of CHROMagar Orientation (CO) medium into routine use in two hospital laboratories and compared it to conventional culture on blood and MacConkey agars. Based on data extracted from our Laboratory Information System from 2006 to 2011, the use of CO medium resulted in a 28% reduction in workload for additional procedures such as Gram stains, subcultures, identification panels, agglutination tests, and biochemical tests. The average number of workload units (one workload unit equals 1 min of hands-on labor) per urine specimen was significantly reduced (P < 0.0001; 95% confidence interval [CI], 0.5326 to 1.047) from 2.67 in 2006 (preimplementation of CO medium) to 1.88 in 2011 (postimplementation of CO medium). We conclude that the use of CO medium streamlined the urine culture process and increased bench throughput by reducing both workload and turnaround time in our laboratories. PMID:23363839

  16. Does culture medium influence offspring birth weight?

    Science.gov (United States)

    Carrasco, Beatriz; Boada, Montserrat; Rodríguez, Ignacio; Coroleu, Buenaventura; Barri, Pedro N; Veiga, Anna

    2013-11-01

    To determine whether the type of medium used to culture human embryos in vitro influences neonatal birth weight after IVF/intracytoplasmic sperm injection (ICSI). A prospective study and a retrospective study. Private assisted reproduction center. The prospective study included 449 IVF/ICSI cycles from August to December 2008. The retrospective analysis was performed for 2,518 IVF/ICSI cycles from October 2006 to December 2010. In the prospective study, patients were randomized for embryo culture in Cook or Vitrolife medium. The retrospective study was performed with three different culture media (MediCult, Cook, and Vitrolife). Mean birth weight, adjusted for gestational age and gender (z score) of newborns. In the prospective study, the average z score was -0.19 ± 0.85 in Cook and 0.08 ± 1.40 in Vitrolife. In the retrospective study, the z scores obtained in each group were as follows: Cook, -0.14 ± 0.96; MediCult, 0.06 ± 1.13; and Vitrolife, 0.03 ± 1.05. No significant differences were observed regarding the birth weight of children born in the different groups in both studies. The results do not show any relationship between the medium used for in vitro culture and mean birth weight adjusted for gestational age and gender of singletons born after IVF/ICSI. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  17. Development of a vinasse culture medium for plant tissue culture

    International Nuclear Information System (INIS)

    Silva, A.L.L.D.; Gollo, L.

    2014-01-01

    Vinasse is the main pollutant (effluent) obtained from the distillation of sugarcane in the production of fuel alcohol. However, this residue is rich in nutrients that are required by plants. We developed a new culture medium using vinasse for the In vitro propagation of an orchid. The vinasse was treated (decanted and filtered), and the nutrients were determined and quantified. Different formulations using vinasse were tested for an In vitro culture. The vinasse dilutions demonstrated a good buffering effect. The ideal vinasse dilution for media formulation was 2.5%. The best KC formulations with vinasse were KCV1 and KCV5. Compared to KC medium, these formulations demonstrated similar results for In vitro multiplication, with the exception of protocorm-like body number, which was inferior in the vinasse formulations. Conversely, for In vitro elongation and rooting, these vinasse media were superior to KC medium. KC medium promotes a low rooting rate (8%) compared to 68 and 100% obtained by KCV1 and KCV5, respectively. Moreover, plantlets cultured on KC medium become protocorm-like body clusters, which impeded the acclimatization of these explants. Plantlets elongated and rooted on KCV1 and KCV5 were successfully acclimatized with a 91% survival rate for both KC vinasse formulations. This study shows the great potential of this technology as a rational alternative to vinasse disposal and adds value to what is currently considered a waste product. (author)

  18. Medium-temperature solid oxide fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Maffei, N.; Kuriakose, A.K. [Natural Resources Canada, Ottawa, ON (Canada). Materials Technology Lab

    2000-07-01

    The Materials Technology Laboratory (MTL) of Natural Resources Canada has been conducting research on the development of a solid oxide fuel cell (SOFC) for the past decade. Fuel cells convert chemical energy directly into electric energy in an efficient and environmentally friendly manner. SOFCs are considered to be good stationary power sources for commercial and residential applications and will likely be commercialized in the near future. The research at MTL has focused on the development of new electrolytes for use in SOFCs. In the course of this research, monolithic planar single cell SOFCs based on doubly doped ceria and lanthanum gallate have been fabricated and tested at 700 degrees C. This paper compared the performance characteristics of both these systems. The data suggested the presence of a significant electronic conductivity in the SOFC incorporating doubly doped ceria, resulting in lower than expected voltage output. The stability of the SOFC, however, did not appear to be negatively affected. The lanthanum gallate based SOFC performed well. It was concluded that reducing the operating temperature of SOFCs would improve their reliability and enhance their operating life. First generation commercial SOFCs will use a zirconium oxide-based electrolytes while second generation units might possibly use ceria-based and/or lanthanum gallate electrolytes. 24 refs., 6 figs.

  19. INNOVATIVE CULTURE IN SMALL AND MEDIUM ENTERPRISES

    Directory of Open Access Journals (Sweden)

    Aluisio Broering Mambrini

    2011-10-01

    Full Text Available In the last two decades, innovation has been a key driver of economic growth. Innovation is closely related to creating value and generating wealth through successful service to consumer needs. Thus, it is not necessarily restricted to the use of new knowledge generated from research, but on the development of new products or services that are obtained with creative use of knowledge, new or already known. This study aimed to identify management practices that promote a culture of innovation in small and medium enterprises and analyze how they contribute to the innovative capacity of these companies. The research method was the multiple case study with six small and medium businesses that have at least one case of significant innovation in its history. The main results showed that amongst the practices are: a performance in highly specialized niches and deep focus on customer needs; b strong investment and incorporation of new knowledge outside the company (open innovation; c speed and agility in the absorption and deployment of new knowledge and technologies; d retention of employees; e acting as an integrator combining diverse knowledge and technologies; f the information management of the knowledge acquired by the company; g little concern to patent the technology; h flexibility and informal, fluid and open communication between employees of the company that promotes agility in management and i the management of partnerships across the value chain, including the functional areas.

  20. Medium optimization for protopectinase production by batch culture of

    African Journals Online (AJOL)

    Medium optimization for protopectinase production by batch culture of. C Fan, Z Liu, L Yao. Abstract. Optimization of medium compositions for protopectinase production by Aspergillus terreus in submerged culture was carried out. The medium components having significant effect on protopectinase production were reported ...

  1. Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.

    Science.gov (United States)

    Lemos, M; Souza, C S F; da Costa, S C Gonçalves; Souto-Padrón, T; D'Agosto, M

    2013-02-01

    The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.

  2. 21 CFR 866.2360 - Selective culture medium.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2360 Selective culture... solid biological materials intended for medical purposes to cultivate and identify certain pathogenic...

  3. 21 CFR 866.2330 - Enriched culture medium.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2330 Enriched culture... solid biological materials intended for medical purposes to cultivate and identify fastidious...

  4. [Influence of liquid or solid culture conditions on the volatile components of mycelia of Isariacateinannulata].

    Science.gov (United States)

    Zhang, Delong; Wang, Xiaodong; Lu, Ruili; Li, Kangle; Hu, Fenglin

    2011-12-01

    To determine the volatile components of mycelia of Isaria cateinannulata cultured under different culture conditions, and to analyze the relationships between the culture conditions and volatile metabolites. Mycelia were cultured in solid plates with SDAY medium and liquid shake flasks with SDY medium. The culture conditions were at 25 degrees C and 8 days. Volatile components in the mycelia of I. cateinannulata were extracted with simultaneous distillation extraction and analyzed by gas chromatography-mass spectrometry. Alkenes, alkanes, heterocyclic and polycyclic aromatic hydrocarbons (PAH) were existed abundantly both in the mycelia of liquid and solid cultures, but the kinds and relative concentrations of the volatile components in mycelia of liquid and solid cultures were very different. Forty-one compounds were identified from the mycelia of solid culture and 32 compounds were identified from the mycelia of liquid culture. Esters, quinones and oximes were only found in solid cultured mycelia whereas carboxylic acids were only discovered in the mycelia of liquid culture. At the same time, mycelia of liquid culture contained much more phenols. The most abundant compounds in mycelia of liquid and solid cultures were hydrocarbons. The volatile extracts of solid cultured mycelia contained 57.6% alkenes and 9.19% alkanes. The volatile extracts of liquid cultured mycelia contained 7.85% alkenes and 22.4% alkanes. Liquid or solid culture conditions influenced the volatile components of mycelia of I. cateinannulata.

  5. Low and medium activity solid wastes processing and encapsulation

    International Nuclear Information System (INIS)

    Taillard, D.; Claes, J.; Hennart, D.

    1983-01-01

    This work, carried out under contract with the European Atomic Energy Community, describes the techniques in use for waste management. The activity of low and medium activity solid wastes is from few curies to few tens of curies per cubic meter, they are produced by nuclear facilities and are often complex mixtures. Radioactive wastes are characterized and processing and conditioning are described. Leaching, stability, mechanical resistance and radiolysis of encapsulated wastes are examined. Handling, storage and disposal are treated

  6. Optimizing culture medium for debittering constitutive enzyme ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-08-02

    Aug 2, 2010 ... naringinase on different matrices has been studied by many researchers (Busto et ... 10 g/L in the base medium compared to naringin control. Nitrogen ... Fermentation experiments were carried out in shaking flask for 5 days at 28°C with initial pH 6.0. † Values ..... fujikuroi mycelium in fluidized bioreactors.

  7. The low and medium level solid radioactive wastes

    International Nuclear Information System (INIS)

    1981-01-01

    One of the most controversial aspects in the nuclear energy utilization for pacific purposes is related with the low and medium level solid radioactive wastes production, during the nuclear power plants operation. These wastes shall be inmobilizated before their storage on-site or their transport out-site. This paper presents an exposition about the avobe mentioned problem, attending three basic areas: the fundamental concepts and criteria for their management, the methods used in western countries and the present situation of the spanish nuclear power plants in operation and under construction. (auth.)

  8. Arsenolite: a quasi-hydrostatic solid pressure-transmitting medium

    International Nuclear Information System (INIS)

    Sans, J A; Manjón, F J; Popescu, C; Muñoz, A; Rodríguez-Hernández, P; Jordá, J L; Rey, F

    2016-01-01

    This study reports the experimental characterization of the hydrostatic properties of arsenolite (As 4 O 6 ), a molecular solid which is one of the softest minerals in the absence of hydrogen bonding. The high compressibility of arsenolite and its stability up to 15 GPa have been proved by x-ray diffraction measurements, and the progressive loss of hydrostaticity with increasing pressure up to 20 GPa has been monitored by ruby photoluminescence. Arsenolite has been found to exhibit hydrostatic behavior up to 2.5 GPa and a quasi-hydrostatic behavior up to 10 GPa at room temperature. This result opens the way to explore other molecular solids as possible quasi-hydrostatic pressure-transmitting media. The validity of arsenolite as an insulating, stable, non-penetrating and quasi-hydrostatic medium is explored by the study of the x-ray diffraction of zeolite ITQ-29 at high pressure. (paper)

  9. Sodium hypochlorite sterilization of culture medium in ...

    African Journals Online (AJOL)

    With the purpose of finding an alternative to thermal sterilization, this research aimed at assessing the efficiency and ideal concentration of sodium hypochlorite for sterilization of culture media and glassware used during rooting of micropropagated Gerbera hybrida cv. Essandre. Two experiments were carried out. In the first ...

  10. 21 CFR 866.2390 - Transport culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Transport culture medium. 866.2390 Section 866.2390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2390 Transport culture...

  11. Architecture of Institution & Home. Architecture as Cultural Medium

    NARCIS (Netherlands)

    Robinson, J.W.

    2004-01-01

    This dissertation addresses how architecture functions as a cultural medium. It does so by by investigating how the architecture of institution and home each construct and support different cultural practices. By studying the design of ordinary settings in terms of how qualitative differences in

  12. Storage facility for solid medium level waste at Eurochemic

    International Nuclear Information System (INIS)

    Balseyro-Castro, M.

    1976-01-01

    An engineered surface storage facility is described; it will serve for the interim storage of solid and solidified medium-level waste resulting from the reprocessing of irradiated fuels. Up till now, two storage bunkers have been constructed. Each of them is 64 m long, 12 m wide and 8 m high and can take up to about 5,000 drums of 220 1 volume. The drums are stored in a vertical position and in four layers. The waste product drums are transported by a wagon to the entrance of the bunkers from where they are transferred in to the bunker by an overhead crane which is remotely controlled by high-frequency modulated laser beams. A closed-circuit camera is used to watch the handling operations. The waste stored is fully retrievable, either by means of an overhead crane of a lift-truck and can then be transported to an ultimate storage site

  13. Culture medium for amylase production by toxigenic fungi

    Directory of Open Access Journals (Sweden)

    Figueira Edson Luiz Zangrando

    2000-01-01

    Full Text Available Mycelial growth and amylase production by a mycotoxigenic strain of Fusarium moniliforme and Aspergillus flavus were evaluated in a culture medium containing starch, glycerol, wheat bran or corn. With emphasis on corn, different fractions composed by germ, degermed seed, starch, milky stage corn and the respective starch or supernatant fraction were analyzed for F. moniliforme growth . The medium composed of milky stage corn supernatant promoted the best mycelial growth (p<0.05, and it was used to prepare amylase production medium in the next step. The medium composed with 2% ground corn in milky stage corn supernatant (350g of milky stage corn blended with 250mL water and centrifuged promoted the highest amylase production, which was at the 10th day of fermentation, both for F. moniliforme (42.32U/mL and A. flavus (4,745.54U/mL.

  14. Liquid vs Solid Culture Medium to Evaluate Proportion and Time to Change in Management of Suspects of Tuberculosis-A Pragmatic Randomized Trial in Secondary and Tertiary Health Care Units in Brazil.

    Directory of Open Access Journals (Sweden)

    Adriana da Silva Rezende Moreira

    Full Text Available The use of liquid medium (MGIT960 for tuberculosis (TB diagnosis was recommended by WHO in 2007. However, there has been no evaluation of its effectiveness on clinically important outcomes.A pragmatic trial was carried out in a tertiary hospital and a secondary health care unit in Rio de Janeiro City, Brazil. Participants were 16 years or older, suspected of having TB. They were excluded if only cerebral spinal fluid or blood specimens were available for analysis. MGIT960 technique was compared with the Lowenstein-Jensen (LJ method for laboratory diagnosis of active TB. Primary outcome was the proportion of patients who had their initial medical management changed within 2 months after randomisation. Secondary outcomes were: mean time for changing the procedure, patient satisfaction with the overall treatment and adverse events. Data were analysed by intention-to-treat. Between April 2008 and September 2011, 693 patients were enrolled (348 to MGIT, 345 to LJ. Smear and culture results were positive for 10% and 15.7% of participants, respectively. Patients in the MGIT arm had their initial medical management changed more frequently than those in the LJ group (10.1% MGIT vs 3.8% LJ, RR 2.67 95% CI 1.44-.96, p = 0.002, NNT 16, 95% CI 10-39. Mean time for changing the initial procedure was greater in LJ group at both sites: 20.0 and 29.6 days in MGIT group and 52.2 and 64.3 in LJ group (MD 33.5, 95% CI 30.6-36.4, p = 0.0001. No other important differences were observed.This study suggests that opting for the MGIT960 system for TB diagnosis provides a promising case management model for improving the quality of care and control of TB.Controlled-Trials.com ISRCTN79888843.

  15. Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium

    Directory of Open Access Journals (Sweden)

    Mitchison Denis A

    2004-11-01

    Full Text Available Abstract Background Old, stationary cultures of Mycobacterium tuberculosis contain a majority of bacteria that can grow in broth cultures but cannot grow on solid medium plates. These may be in a non-replicating, dormant growth phase. We hypothesised that a similar population might be present in chronic, murine tuberculosis. Methods Estimates of the numbers of viable M. tuberculosis, strain H37Rv, in the spleens and lungs of mice in a 7-day acute infection and in a 10-month chronic infection were made by conventional plate counts and, as broth counts, by noting presence or absence of growth in serial replicate dilutions in liquid medium. Results Plate and broth counts in 6 mice gave similar mean values in the acute infection, 7 days after infection. However, the broth counts were much higher in 36 mice with a chronic infection at 10 months. Broth counts averaged 5.290 log10 cfu /organ from spleens and 5.523 log10 cfu/organ from lungs, while plate counts were 3.858 log10 cfu/organ from spleens and 3.662 log10 cfu/organ from lungs, indicating that the total bacterial population contained only 3.7% bacilli in spleens and 1.4% bacilli in lungs, capable of growth on plates. Conclusion The proportion growing on plates might be a measure of the "dormancy" of the bacilli equally applicable to cultural and animal models.

  16. Epileptogenesis in organotypic hippocampal cultures has limited dependence on culture medium composition.

    Directory of Open Access Journals (Sweden)

    Jing Liu

    Full Text Available Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium, which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF. Therefore, it is possible that epileptogenesis in organotypic cultures is driven by these components. We examined the influence of medium composition on epileptogenesis. Epileptogenesis was evaluated by measurements of lactate and lactate dehydrogenase (LDH levels (biomarkers of ictal activity and cell death, respectively in spent culture media, immunohistochemistry and automated 3-D cell counts, and extracellular recordings from CA3 regions. Changes in culture medium components moderately influenced lactate and LDH levels as well as electrographic seizure burden and cell death. However, epileptogenesis occurred in any culture medium that was capable of supporting neural survival. We conclude that medium composition is unlikely to be the cause of epileptogenesis in the organotypic hippocampal culture model of chronic post-traumatic epilepsy.

  17. Epileptogenesis in organotypic hippocampal cultures has limited dependence on culture medium composition.

    Science.gov (United States)

    Liu, Jing; Saponjian, Yero; Mahoney, Mark M; Staley, Kevin J; Berdichevsky, Yevgeny

    2017-01-01

    Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium), which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF). Therefore, it is possible that epileptogenesis in organotypic cultures is driven by these components. We examined the influence of medium composition on epileptogenesis. Epileptogenesis was evaluated by measurements of lactate and lactate dehydrogenase (LDH) levels (biomarkers of ictal activity and cell death, respectively) in spent culture media, immunohistochemistry and automated 3-D cell counts, and extracellular recordings from CA3 regions. Changes in culture medium components moderately influenced lactate and LDH levels as well as electrographic seizure burden and cell death. However, epileptogenesis occurred in any culture medium that was capable of supporting neural survival. We conclude that medium composition is unlikely to be the cause of epileptogenesis in the organotypic hippocampal culture model of chronic post-traumatic epilepsy.

  18. Aleuria aurantia - indole metabolites of fruit bodies, mycelial culture and culture medium

    Directory of Open Access Journals (Sweden)

    Janina Węgiel

    2014-08-01

    Full Text Available The aim of present study was to investigate and compare indole metabolites of fruit bodies, mycelium cultivated in vitro and culture medium of the fungus Aleuria aurantia (Fr. Fuck. By use of a number of chromatographic and spectroscopic methods several indole metabolites have been detected and identified among other the 3-indolebutyric acid was produced and extracted to the culture medium. Furthermore 3-indoleatonitrile and tryptophane degradative products have been found both in fruit bodies and mycelium.

  19. Sugarcane Bagasse: A Potential Medium for Fungal Cultures

    OpenAIRE

    Arushdeep Sidana; Umar Farooq

    2014-01-01

    Worldwide, sugarcane industries produce tons of sugarcane bagasse as residual/waste material. This residual material is rich in complex lignocellulosic substances and may be used as a low cost carbon and energy source for the growth of fungal species. The present work was aimed at designing a sugarcane waste-based medium as a substitute for expensive commercial media for growing fungal cultures. Eight species of fungi, namely, Aspergillus niger, Candida albicans, Saccharomyces cerevisiae, Fus...

  20. Determination of Glucose Concentration in Yeast Culture Medium

    Science.gov (United States)

    Hara, Seiichi; Kishimoto, Tomokazu; Muraji, Masafumi; Tsujimoto, Hiroaki; Azuma, Masayuki; Ooshima, Hiroshi

    The present paper describes a sensor for measuring the glucose concentration of yeast culture medium. The sensor determines glucose concentration by measuring the yield of hydrogen peroxide produced by glucose oxidase, which is monitored as luminescence using photomultiplier. The present sensor is able to measure low glucose concentration in media in which yeast cells keep respiration state. We herein describe the system and the characteristics of the glucose sensor.

  1. Morphological differentiation of Streptomyces viridochromogenes E-219 on solid culture

    International Nuclear Information System (INIS)

    Liang Xinle; Zhu Jing; Jin Yingyan

    2012-01-01

    The Streptomyces viridochromogenes E-219 was derived from Streptomyces viridochromogenes CGMCC4.1119 treated with 60 Co γ-rays irradiation and protoplast fusion. With the help of fluorescent probes, fluorescence microscope and electron microscopy, the morphology and development of E-219 on solid surface culture were investigated in this study. The effect of agarslant culture time on the production of Avilamycin was also studied to provide theoretical basis for industrial fermentation of selecting the appropriate seed to culture on the agarslant culture medium. The results implied that the development of colonies of Streptomyces viridochromogenes accompanied the intermittent hyhae apoptosis, and the production of spores was from the active mycelium. The colonial morphology of strain E-219 was significantly different from the original strain CGMCC4h1119. There were variegated hyphae formation in the stage of spore germination and initial hyphae development (10 h) with the live and dead segments alternated in a highly regular fashion within the same hypha. After the early single colony formation, the third phase was followed by profuse growth of the live segments derived from the variegated hypha, then the second apoptosis of the mycelia (48 h) was occurred with another quick growth, and sporulation was occurred at 96 h. Strain CGMCC4.1119 had spiral sporotrichial and round conidiophores with spike, whereas strain E-219 had linear sporotrichial, smooth and dylindrical conidiophore. The results of shake flask experiments indicated that the spores of E-219 had that highest activity when cultured on agarslant culture medium and incubated for 106 h with the production of avilamycin up to 1200 mg/L. (authors)

  2. Effect of culture medium on propagation and phenotype of corneal stroma-derived stem cells.

    Science.gov (United States)

    Sidney, Laura E; Branch, Matthew J; Dua, Harminder S; Hopkinson, Andrew

    2015-12-01

    The limbal area of the corneal stroma has been identified as a source of mesenchymal-like stem cells, which have potential for exploitation as a cell therapy. However, the optimal culture conditions are disputed and few direct media comparisons have been performed. In this report, we evaluated several media types to identify the optimal for inducing an in vitro stem cell phenotype. Primary human corneal stroma-derived stem cells (CSSCs) were extracted from corneoscleral rims. Culture in seven different media types was compared: Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS); M199 with 20% FBS; DMEM-F12 with 20% serum replacement, basic fibroblast growth factor and leukemia inhibitory factor (SCM); endothelial growth medium (EGM); semi-solid MethoCult; serum-free keratinocyte medium (K-SFM); and StemPro-34. Effects on proliferation, morphology, protein and messenger RNA expression were evaluated. All media supported proliferation of CSSCs with the exception of K-SFM and StemPro-34. Morphology differed between media: DMEM produced large cells, whereas EGM produced very small cells. Culture in M199 produced a typical mesenchymal stromal cell phenotype with high expression of CD105, CD90 and CD73 but not CD34. Culture in SCM produced a phenotype more reminiscent of a progenitor cell type with expression of CD34, ABCG2, SSEA-4 and PAX6. Culture medium can significantly influence CSSC phenotype. SCM produced a cell phenotype closest to that of a pluripotent stem cell, and we consider it to be the most appropriate for development as a clinical-grade medium for the production of CSSC phenotypes suitable for cell therapy. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  3. Morphology of primary human venous endothelial cell cultures before and after culture medium exchange.

    Science.gov (United States)

    Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P

    2015-01-01

    The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the

  4. Developing a Plant Culture Medium Composed of Vinasse Originating from Haematococcus Pluvialis Culture

    International Nuclear Information System (INIS)

    Gollo, A. L.; Silva, A. L. L. D.; Lima, K. K. D. D.; Camara, M. C.; Rodrigues, C.; Vandenberghe, L. P. D. S.; Soccol, V. T.; Soccol, C. R.; Biasi, L. A.

    2016-01-01

    The mineral nutrients in vinasse provide support for algal and plant growth. Algal culture releases organic compounds into its liquid culture medium. These organic and inorganic substances can be useful for formulating a plant tissue culture medium, because tissue culture medium is composed of organic and inorganic components. Therefore, the aims of this study were to develop a plant culture medium by using the vinasse that is employed for Haematococcus pluvialis culture (algal filtrate); to investigate the possible beneficial effects of the biocompounds in the micropropagation of Nidularium procerum (Bromeliaceae), to evaluate quercetin content, total phenolics content in vinasse and to evaluate the cytotoxicity of the media by performing a bioassay with Artemia salina. The vinasse that originated from H. pluvialis culture can be used to formulate plant tissue culture at a 3% dilution, and its mineral nutrients can support In vitro plant growth, but some nutrients must be supplemented to enhance its efficiency. An efficient micropropagation protocol was developed for N. procerum. The micropropagated plants were suitable for transfer to the field (they were acclimatized). This culture medium provides a way to reuse wastewater, gives a rational alternative to vinasse disposal and adds value to what is currently considered to be an undesirable residue. Moreover, this process can reduce the production costs of clonal seedlings and/or bioactive compounds in biofactories. There was no apparent biostimulatory effect by the algal filtrate on morphogenesis; however, it did increase quercetin production. The H. pluvialis culture that was grown in the vinasse decreased the cytotoxicity and phenolic compound contents, which prevented explant tissue necrosis and represented a treatment for this residue for safer disposal in the environment. (author)

  5. Effective medium of periodic fluid-solid composites

    KAUST Repository

    Mei, Jun; Wu, Ying; Liu, Zhengyou

    2012-01-01

    An analytic solution of the effective mass density and bulk modulus of a periodic fluid-solid composite is obtained by using the multiple-scattering theory in the long-wavelength limit. It is shown that when the concentration of solid inclusions

  6. Epileptogenesis in organotypic hippocampal cultures has limited dependence on culture medium composition

    OpenAIRE

    Liu, Jing; Saponjian, Yero; Mahoney, Mark M.; Staley, Kevin J.; Berdichevsky, Yevgeny

    2017-01-01

    Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium), which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF). Therefore, it is possible that epileptogenesis in organotypic ...

  7. Selection of culture medium and conditions for the production of ...

    African Journals Online (AJOL)

    defined medium–A, defined medium-B, synthetic medium, rich medium and industrial medium) showed that the synthetic medium yielded maximum yeast biomass (12.8 g/LDCW) followed by rich medium (11.7 g/L DCW) and defined medium B ...

  8. Rapid Isolation and Susceptibility Testing of Leptospira spp. Using a New Solid Medium, LVW Agar

    Science.gov (United States)

    Wuthiekanun, Vanaporn; Amornchai, Premjit; Paris, Daniel H.; Langla, Sayan; Thaipadunpanit, Janjira; Chierakul, Wirongrong; Smythe, Lee D.; White, Nicholas J.; Day, Nicholas P. J.; Peacock, Sharon J.

    2013-01-01

    Pathogenic Leptospira spp., the causative agents of leptospirosis, are slow-growing Gram-negative spirochetes. Isolation of Leptospira from clinical samples and testing of antimicrobial susceptibility are difficult and time-consuming. Here, we describe the development of a new solid medium that facilitates more-rapid growth of Leptospira spp. and the use of this medium to evaluate the Etest's performance in determining antimicrobial MICs to drugs in common use for leptospirosis. The medium was developed by evaluating the effects of numerous factors on the growth rate of Leptospira interrogans strain NR-20157. These included the type of base agar, the concentration of rabbit serum (RS), and the concentration and duration of CO2 incubation during the initial period of culture. The highest growth rate of NR-20157 was achieved using a Noble agar base supplemented with 10% RS (named LVW agar), with an initial incubation at 30°C in 5% CO2 for 2 days prior to continuous culture in air at 30°C. These conditions were used to develop the Etest for three species, L. interrogans (NR-20161), L. kirschnerii (NR-20327), and L. borgpetersenii (NR-20151). The MICs were read on day 7 for all samples. The Etest was then performed on 109 isolates of pathogenic Leptospira spp. The MIC90 values for penicillin G, doxycycline, cefotaxime, ceftriaxone, and chloramphenicol were 0.64 units/ml and 0.19, 0.047, 0.5, and 2 μg/ml, respectively. The use of LVW agar, which enables rapid growth, isolation of single colonies, and simple antimicrobial susceptibility testing for Leptospira spp., provides an opportunity for new areas of fundamental and applied research. PMID:23114772

  9. Effective medium of periodic fluid-solid composites

    KAUST Repository

    Mei, Jun

    2012-05-24

    An analytic solution of the effective mass density and bulk modulus of a periodic fluid-solid composite is obtained by using the multiple-scattering theory in the long-wavelength limit. It is shown that when the concentration of solid inclusions is high, the effective mass density is structure dependent and differs significantly from the leading-order dipole solution, whereas Wood\\'s formula is accurately valid, independently of the structures. Numerical evaluations from the analytic solution are shown to be in excellent agreement with finite-element simulations. In the vicinity of the tight-packing limit, the critical behavior of the effective mass density is also studied and it is independent of the lattice symmetry. © 2012 Europhysics Letters Association.

  10. Mathematical modeling of ethanol production in solid-state fermentation based on solid medium' dry weight variation.

    Science.gov (United States)

    Mazaheri, Davood; Shojaosadati, Seyed Abbas; Zamir, Seyed Morteza; Mousavi, Seyyed Mohammad

    2018-04-21

    In this work, mathematical modeling of ethanol production in solid-state fermentation (SSF) has been done based on the variation in the dry weight of solid medium. This method was previously used for mathematical modeling of enzyme production; however, the model should be modified to predict the production of a volatile compound like ethanol. The experimental results of bioethanol production from the mixture of carob pods and wheat bran by Zymomonas mobilis in SSF were used for the model validation. Exponential and logistic kinetic models were used for modeling the growth of microorganism. In both cases, the model predictions matched well with the experimental results during the exponential growth phase, indicating the good ability of solid medium weight variation method for modeling a volatile product formation in solid-state fermentation. In addition, using logistic model, better predictions were obtained.

  11. Cultivo inicial in vitro de gemas axilares de Ananas comosus (L. Merr., em meio líquido/sólido, na presença/ausência de luz In nitial in vitro culture of axillary buds of Ananas comosus (L. Merr. in liquid/semi-solid culture medium, in the presence/absence of light

    Directory of Open Access Journals (Sweden)

    Sarah Brandão Santa Cruz Barboza

    2009-01-01

    Full Text Available A capacidade regenerativa de gemas axilares de abacaxizeiro cv. Pérola (Ananas comosus (L. Merr. foi avaliada em meio de cultivo líquido e gelificado, na presença e ausência de luz, com o objetivo de produzir material para subsequentes fases da micropropagação. Os explantes foram estabelecidos, por 60 dias, em meio MS acrescido de 0,63 μM de ANA e 2,5 μM de BAP. O delineamento experimental foi inteiramente casualizado, constituído com quatro tratamentos (meio líquido com e sem ponte de papel de filtro e meio gelificado com ágar a 6,0 g L-1 e com Phytagel® a 3,0 g L-1, com quatro repetições e cinco explantes por repetição. Diferentes volumes de meio líquido (2,5 mL; 5,0 mL e 10,0 mL também foram avaliados, usando-se quatro repetições por volume de meio de cultura. Maior número de brotos foi obtido na presença de luz, em meio líquido sem ponte de papel de filtro, onde 80% dos explantes se diferenciaram em brotos. Na ausência de luz, 68,8% a 87,5% dos explantes não regeneraram brotos. Explantes cultivados em volumes de 5,0 mL ou 2,5 mL de meio MS líquido, acrescido de 0,63 μM de ANA e 2,5 μM de BAP, na presença de luz, constitui um protocolo adequado para cultivo inicial de gemas axilares e regeneração de brotos de abacaxizeiro.The regenerative capacity of buds from lateral sprouts of the pineapple cv. Pérola (Ananas comosus (L. Merr. was evaluated in liquid and semi-solid medium, under light and dark conditions, with the objective of producing materials for subsequent phase of micropropagation. The explants were established for 60 days in basal MS medium, supplemented with 0.63 μM NAA and 2.5 μM BAP. The statistical design was completely randomized with four treatments (liquid medium with and without filter paper bridge, and semi-solid medium gellified with agar and phytagel®, four repetitions and five explants per experimental unit. Different volumes of liquid medium (2.5 mL, 5.0 mL, and 10.0 mL were also

  12. [Culture conditions for gametes and embryos: Which culture medium? Which impact on newborn?

    Science.gov (United States)

    Koscinski, I; Merten, M; Kazdar, N; Guéant, J-L

    2018-05-01

    Many studies have examined the impact of cell/embryo culture media on the development of human embryo during IVF process, but few studies have followed up and compared the effects of these culture media on the developmental outcome of children conceived by IVF. As recurrent experimental evidence from animal studies suggests potential long-term effects of embryo culture media on the health outcome of IVF-conceived children, more studies are needed to clarify the role of the culture media and mechanisms underlying such effects. In human, however, the effects of culture media are difficult to pinpoint due to complications stem from both the influence of maternal nutrition during the gestational period and the parental genetic. Based on a simple review of the literature integrating animal experimentations and human clinic studies, we suggest that the composition of culture medium should be considered beyond the character of unique or sequential medium, corresponding to "let embryo choose" or "back to nature" respectively. Instead, we suggest that the main components of embryo culture media should be considered from the point of view of metabolic consequences and potential epigenetic effects. Given that energetic metabolites can regulate epigenetic machinery, we hypothesize that metabolic abnormalities linked to morphological abnormalities could reveal epigenetic defects in embryos. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  13. Protective layer formation on magnesium in cell culture medium

    Energy Technology Data Exchange (ETDEWEB)

    Wagener, V.; Virtanen, S., E-mail: virtanen@ww.uni-erlangen.de

    2016-06-01

    In the past, different studies showed that hydroxyapatite (HA) or similar calcium phosphates can be precipitated on Mg during immersion in simulated body fluids. However, at the same time, in most cases a dark grey or black layer is built under the white HA crystals. This layer seems to consist as well of calcium phosphates. Until now, neither the morphology nor its influence on Mg corrosion have been investigated in detail. In this work commercially pure magnesium (cp) was immersed in cell culture medium for one, three and five days at room temperature and in the incubator (37 °C, 5% CO{sub 2}). In addition, the influence of proteins on the formation of a corrosion layer was investigated by adding 20% of fetal calf serum (FCS) to the cell culture medium in the incubator. In order to analyze the formed layers, SEM images of cross sections, X-ray Photoelectron Spectroscopy (XPS), X-ray diffraction (XRD), Energy Dispersive X-ray Spectroscopy (EDX) and Fourier Transformed Infrared Spectroscopy (FTIR) measurements were carried out. Characterization of the corrosion behavior was achieved by electrochemical impedance spectroscopy (EIS) and by potentio-dynamic polarization in Dulbecco's Modified Eagle's Medium (DMEM) at 37 °C. Surface analysis showed that all formed layers consist mainly of amorphous calcium phosphate compounds. For the immersion at room temperature the Ca/P ratio indicates the formation of HA, while in the incubator probably pre-stages to HA are formed. The different immersion conditions lead to a variation in layer thicknesses. However, electrochemical characterization shows that the layer thickness does not influence the corrosion resistance of magnesium. The main influencing factor for the corrosion behavior is the layer morphology. Thus, immersion at room temperature leads to the highest corrosion protection due to the formation of a compact outer layer. Layers formed in the incubator show much worse performances due to completely porous

  14. Protective layer formation on magnesium in cell culture medium.

    Science.gov (United States)

    Wagener, V; Virtanen, S

    2016-06-01

    In the past, different studies showed that hydroxyapatite (HA) or similar calcium phosphates can be precipitated on Mg during immersion in simulated body fluids. However, at the same time, in most cases a dark grey or black layer is built under the white HA crystals. This layer seems to consist as well of calcium phosphates. Until now, neither the morphology nor its influence on Mg corrosion have been investigated in detail. In this work commercially pure magnesium (cp) was immersed in cell culture medium for one, three and five days at room temperature and in the incubator (37 °C, 5% CO2). In addition, the influence of proteins on the formation of a corrosion layer was investigated by adding 20% of fetal calf serum (FCS) to the cell culture medium in the incubator. In order to analyze the formed layers, SEM images of cross sections, X-ray Photoelectron Spectroscopy (XPS), X-ray diffraction (XRD), Energy Dispersive X-ray Spectroscopy (EDX) and Fourier Transformed Infrared Spectroscopy (FTIR) measurements were carried out. Characterization of the corrosion behavior was achieved by electrochemical impedance spectroscopy (EIS) and by potentio-dynamic polarization in Dulbecco's Modified Eagle's Medium (DMEM) at 37°C. Surface analysis showed that all formed layers consist mainly of amorphous calcium phosphate compounds. For the immersion at room temperature the Ca/P ratio indicates the formation of HA, while in the incubator probably pre-stages to HA are formed. The different immersion conditions lead to a variation in layer thicknesses. However, electrochemical characterization shows that the layer thickness does not influence the corrosion resistance of magnesium. The main influencing factor for the corrosion behavior is the layer morphology. Thus, immersion at room temperature leads to the highest corrosion protection due to the formation of a compact outer layer. Layers formed in the incubator show much worse performances due to completely porous structures. The

  15. Protective layer formation on magnesium in cell culture medium

    International Nuclear Information System (INIS)

    Wagener, V.; Virtanen, S.

    2016-01-01

    In the past, different studies showed that hydroxyapatite (HA) or similar calcium phosphates can be precipitated on Mg during immersion in simulated body fluids. However, at the same time, in most cases a dark grey or black layer is built under the white HA crystals. This layer seems to consist as well of calcium phosphates. Until now, neither the morphology nor its influence on Mg corrosion have been investigated in detail. In this work commercially pure magnesium (cp) was immersed in cell culture medium for one, three and five days at room temperature and in the incubator (37 °C, 5% CO_2). In addition, the influence of proteins on the formation of a corrosion layer was investigated by adding 20% of fetal calf serum (FCS) to the cell culture medium in the incubator. In order to analyze the formed layers, SEM images of cross sections, X-ray Photoelectron Spectroscopy (XPS), X-ray diffraction (XRD), Energy Dispersive X-ray Spectroscopy (EDX) and Fourier Transformed Infrared Spectroscopy (FTIR) measurements were carried out. Characterization of the corrosion behavior was achieved by electrochemical impedance spectroscopy (EIS) and by potentio-dynamic polarization in Dulbecco's Modified Eagle's Medium (DMEM) at 37 °C. Surface analysis showed that all formed layers consist mainly of amorphous calcium phosphate compounds. For the immersion at room temperature the Ca/P ratio indicates the formation of HA, while in the incubator probably pre-stages to HA are formed. The different immersion conditions lead to a variation in layer thicknesses. However, electrochemical characterization shows that the layer thickness does not influence the corrosion resistance of magnesium. The main influencing factor for the corrosion behavior is the layer morphology. Thus, immersion at room temperature leads to the highest corrosion protection due to the formation of a compact outer layer. Layers formed in the incubator show much worse performances due to completely porous structures. The

  16. Biodegradation of low-density polyethylene (LDPE) by isolated fungi in solid waste medium

    International Nuclear Information System (INIS)

    Zahra, Sahebnazar; Abbas, Shojaosadati Seyed; Mahsa, Mohammad-Taheri; Mohsen, Nosrati

    2010-01-01

    In this study, biodegradation of low-density polyethylene (LDPE) by isolated landfill-source fungi was evaluated in a controlled solid waste medium. The fungi, including Aspergillus fumigatus, Aspergillus terreus and Fusarium solani, were isolated from samples taken from an aerobic aged municipal landfill in Tehran. These fungi could degrade LDPE via the formation of a biofilm in a submerged medium. In the sterilized solid waste medium, LPDE films were buried for 100 days in a 1-L flask containing 400 g sterile solid waste raw materials at 28 deg. C. Each fungus was added to a separate flask. The moisture content and pH of the media were maintained at the optimal levels for each fungus. Photo-oxidation (25 days under UV-irradiation) was used as a pretreatment of the LDPE samples. The progress of the process was monitored by measurement of total organic carbon (TOC), pH, temperature and moisture. The results obtained from monitoring the process using isolated fungi under sterile conditions indicate that these fungi are able to grow in solid waste medium. The results of FT-IR and SEM analyses show that A. terreus and A. fumigatus, despite the availability of other organic carbon of materials, could utilize LDPE as carbon source. While there has been much research in the field of LDPE biodegradation under solid conditions, this is the first report of degradation of LDPE by A. fumigatus.

  17. Probing the mechanisms of drug release from amorphous solid dispersions in medium-soluble and medium-insoluble carriers.

    Science.gov (United States)

    Sun, Dajun D; Lee, Ping I

    2015-08-10

    The objective of the current study is to mechanistically differentiate the dissolution and supersaturation behaviors of amorphous drugs from amorphous solid dispersions (ASDs) based on medium-soluble versus medium-insoluble carriers under nonsink dissolution conditions through a direct head-to-head comparison. ASDs of indomethacin (IND) were prepared in several polymers which exhibit different solubility behaviors in acidic (pH1.2) and basic (pH7.4) dissolution media. The selected polymers range from water-soluble (e.g., PVP and Soluplus) and water-insoluble (e.g., ethylcellulose and Eudragit RL PO) to those only soluble in an acidic or basic dissolution medium (e.g., Eudragit E100, Eudragit L100, and HPMCAS). At 20wt.% drug loading, DSC and powder XRD analysis confirmed that the majority of incorporated IND was present in an amorphous state. Our nonsink dissolution results confirm that whether the carrier matrix is medium soluble determines the release mechanism of amorphous drugs from ASD systems which has a direct impact on the rate of supersaturation generation, thus in turn affecting the evolution of supersaturation in amorphous systems. For example, under nonsink dissolution conditions, the release of amorphous IND from medium-soluble carriers is governed by a dissolution-controlled mechanism leading to an initial surge of supersaturation followed by a sharp decline in drug concentration due to rapid nucleation and crystallization. In contrast, the dissolution of IND ASD from medium-insoluble carriers is more gradual as drug release is regulated by a diffusion-controlled mechanism by which drug supersaturation is built up gradually and sustained over an extended period of time without any apparent decline. Since several tested carrier polymers can be switched from soluble to insoluble by simply changing the pH of the dissolution medium, the results obtained here provide unequivocal evidence of the proposed transition of kinetic solubility profiles from the

  18. Two-stage medium chain fatty acid (MCFA) production from municipal solid waste and ethanol

    NARCIS (Netherlands)

    Grootscholten, T.I.M.; Strik, D.P.B.T.B.; Steinbusch, K.J.J.; Buisman, C.J.N.; Hamelers, B.

    2014-01-01

    Chain elongation is an anaerobic fermentation that produces medium chain fatty acids (MCFAs) from volatile fatty acids and ethanol. These MCFAs can be used as biochemical building blocks for fuel production and other chemical processes. Producing MCFAs from the organic fraction of municipal solid

  19. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Science.gov (United States)

    2010-04-01

    ... tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1700 Culture medium for antimicrobial susceptibility tests. (a) Identification. A culture medium for...

  20. 21 CFR 866.2350 - Microbiological assay culture medium.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2350... consists primarily of liquid or solid biological materials intended for medical purposes to cultivate...

  1. Does the type of culture medium used influence birthweight of children born after IVF?

    Science.gov (United States)

    Zandstra, Heleen; Van Montfoort, Aafke P A; Dumoulin, John C M

    2015-03-01

    Do culture media influence birthweight of children born after IVF? Some studies have observed a significant effect of culture media on birthweight, while others have not, but since most studies compared different culture media, conventional meta-analysis was not possible. Animal studies suggest that in vitro culture of embryos can have a significant effect on the birthweight of offspring when compared with in vivo developed embryos. The type of culture medium (or certain components of the medium) used is one of the causal factors. We reviewed all available literature reporting on a relation between culture medium and birthweight in human studies and a selection of animal studies. An extensive literature search on Pubmed and Medline was performed with relevant search criteria relating to IVF, birthweight and culture medium. Eleven studies reporting on a relationship between culture medium and birthweight in human were included in this review. Five of these found significant differences in birthweight when offspring born after culture in different culture media were compared. The remaining studies did not find differences in birthweight after changing culture medium. The number of human studies is limited and different culture media with different compositions are compared which makes a comparison between the studies difficult, if not impossible. Furthermore, most study designs were retrospective with consecutive use of different culture media and limited sample sizes, which makes bias of the results likely. If it could be confirmed that the type of culture medium used does indeed influence phenotypic characteristics (such as birthweight) of children born after IVF, it would underline the importance of monitoring the health of IVF children in relation to aspects of the laboratory techniques used during embryo culture. No funding was applicable to this study. No conflict of interest is declared. © The Author 2015. Published by Oxford University Press on behalf of the

  2. The culture of Chlorella vulgaris in a recycled supernatant: Effects on biomass production and medium quality

    KAUST Repository

    Hadj-Romdhane, F.; Zheng, Xing; Jaouen, Pascal; Pruvost, Jé ré my; Grizeau, Dominique; Croue, Jean-Philippe; Bourseau, Patrick

    2013-01-01

    Reusing supernatant of microalgae culture medium can have inhibitory or toxic effects on the biomass production because of the release of organic metabolites by cells in the culture medium during their growth. This work investigated the impact of Chlorella vulgaris medium recycling on culture productivity, cells quality and accumulation of excreted metabolites in the culture medium. No significant impact on the C. vulgaris growth was observed after 63days of recycling, the productivity remained stable at around 0.55kgm-3day-1. Organic matters accumulated in supernatant were identified as biopolymers (BP) poor in nitrogen and with a size above 40kDa (probably polysaccharides), and small organic molecules (SOM) richer in nitrogen with a molecular size ranging from 1 to 3kDa. The concentration of biopolymers in the supernatant increased till to a maximum and then decreased, possibly consumed by bacteria, whereas small organic compounds accumulated in the medium. © 2013 Elsevier Ltd.

  3. The culture of Chlorella vulgaris in a recycled supernatant: Effects on biomass production and medium quality

    KAUST Repository

    Hadj-Romdhane, F.

    2013-03-01

    Reusing supernatant of microalgae culture medium can have inhibitory or toxic effects on the biomass production because of the release of organic metabolites by cells in the culture medium during their growth. This work investigated the impact of Chlorella vulgaris medium recycling on culture productivity, cells quality and accumulation of excreted metabolites in the culture medium. No significant impact on the C. vulgaris growth was observed after 63days of recycling, the productivity remained stable at around 0.55kgm-3day-1. Organic matters accumulated in supernatant were identified as biopolymers (BP) poor in nitrogen and with a size above 40kDa (probably polysaccharides), and small organic molecules (SOM) richer in nitrogen with a molecular size ranging from 1 to 3kDa. The concentration of biopolymers in the supernatant increased till to a maximum and then decreased, possibly consumed by bacteria, whereas small organic compounds accumulated in the medium. © 2013 Elsevier Ltd.

  4. 21 CFR 866.2300 - Multipurpose culture medium.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2300 Multipurpose culture... several types of pathogenic microorganisms without the need of additional nutritional supplements. Test...

  5. Production of Mushroom Mycelium as a Protein and Fat Source in Submerged Culture in Medium of Vinasse

    Science.gov (United States)

    Falanghe, H.

    1962-01-01

    Of ten mushroom cultures investigated, only Agaricus campestris, Boletus indecisus, and Tricholoma nudum were capable of growing in submerged culture in medium of vinasse with added salts. Higher fermentative efficiencies were found under these conditions than in medium containing molasses or waste sulfite liquor. A. campestris showed a better capacity to produce protein but, since B. indecisus is capable of developing greater mycelium weight, its fermentative efficiencies are comparable. Both microorganisms could be grown in medium of vinasse with greatly varied amounts, producing higher mycelial weight in media with greater vinasse. The capacity of B. indecisus and A. campestris to utilize the noncarbohydrate fraction in total solids, instead of the total carbohydrates when they are in smaller amount, was observed in medium containing vinasse. B. indecisus and A. campestris were easily separated by filtration from the medium, although T. nudum was difficult to separate by this procedure. In experiments with A. campestris, the adaptative capacity of the organism to vinasse was demonstrated. PMID:13962715

  6. The incorporation of low and medium level radioactive wastes (solids and liquids) in cement

    International Nuclear Information System (INIS)

    Palmer, J.D.; Smith, D.L.G.

    1986-01-01

    The use of cement has been investigated for the immobilization of liquid and solid low and medium level radioactive waste. 220 litre mixing trials have demonstrated that the high temperatures generated during the setting of ordinary Portland cement/simulant waste mixes can be significantly reduced by the use of a blend of ground granulated blast furnace slag and ordinary Portland cement. Laboratory and 220 litre trials using simulant wastes showed that the blended cement gave an improvement in properties of the cemented waste product, e.g. stability and reduction in leach rates compared with ordinary Portland cement formulations. A range of 220 litre scale mixing systems for the incorporation of liquid and solid wastes in cement was investigated. The work has confirmed that cement-based processes can be used for the immobilization of most types of low and medium level waste

  7. Effective medium approximation for elastic constants of porous solids with microscopic heterogeneity

    International Nuclear Information System (INIS)

    Berryman, J.G.

    1986-01-01

    Formulas for the scattering from an inhomogeneous sphere in a fluid-saturated porous medium are used to construct a self-consistent effective medium approximation for the coefficients in Biot's equations of poroelasticity [J. Acoust. Soc. Am. 28, 168 (1956)] when the material constituting the porous solid frame is not homogeneous on the microscopic scale. The discussion is restricted to porous materials exhibiting both macroscopic and microscopic isotropy. Brown and Korringa [Geophysics 40, 608 (1975)] have previously found the general form of these coefficients. The present results give explicit estimates of all the coefficients in terms of the moduli of the solid constituents. The results are also shown to be completely consistent with the well-known results of Gassmann and of Biot and Willis, as well as those of Brown and Korringa

  8. Optimization of an effective growth medium for culturing probiotic bacteria for applications in strict vegetarian food products

    Directory of Open Access Journals (Sweden)

    Manju Pathak

    2012-10-01

    Full Text Available Background: This study aimed to modify de Man Rogosa Sharpe culture medium (termed MRS for selective cultivation of probiotics strain for the consumption by the strictly vegetarian human population. Vegetarian probiotic foods by definition must be free from all animal-derived ingredients. This not only includes the product ingredients but the probiotic inoculum as well. Probiotic starter cultures are traditionally grown and stored in media containing milk or meatderived ingredients. The presence of these ingredients makes the probiotic cell concentrates unsuitable for use in vegetarian products and thus creates the need for a growth medium which isfree from animal-derived ingredients. Present study investigated the growth of a strain of Lactobacillus lactis in MRS. The present invention relates in general to a bacterial culture media,and more specifically a complex microbial culture media, based on plant seed powder extract in place of animal extract for probiotic bacterial growth.Methods: Lactobacillus lactis, a probiotic, was grown in standard MRS culture medium as well as in our various test media (TM containing various vegetal source in place of beef extract, yeast extract and peptone as in case of MRS. The inoculated culture mediums were incubated at 37C for 72 hours and growth of probiotic is recorded at regular intervals. The growth was recorded as Colony Forming Units (CFUs.Results: The best growth of probiotic is observed in TM 2. TM 2 is the leguminous seed extract. Starter culture mediums for probiotics or other bacteria primarily contain protein from animal source. The possibility of using vegetal protein from TM 2 extract in place of peptones and meat extract for the nitrogen supplementation of culture media for the growth of lactic acid bacteria has been demonstrated.Functional Foods in Health and Disease 2012, 2(10:369-378 Conclusion: The absolute vegetarian culture medium containing TM 2 is better than standard MRS for the

  9. Medium optimization for protopectinase production by batch culture ...

    African Journals Online (AJOL)

    ajl yemi

    2011-11-11

    Nov 11, 2011 ... Optimization of medium compositions for protopectinase production by ... food industry, pharmacy and cosmetic manufacture due to ... energy intensive and industrial wastes (Iglesias and ... group of enzymes, which produce the enzymatic ... development time and overall costs (Pan et al., 2008; Ren.

  10. The dynamics of the water droplet impacting onto hot solid surfaces at medium Weber numbers

    Science.gov (United States)

    Mitrakusuma, Windy H.; Kamal, Samsul; Indarto; Dyan Susila, M.; Hermawan; Deendarlianto

    2017-10-01

    The effects of the wettability of a droplet impacting onto a hot solid surface under medium Weber numbers were studied experimentally. The Weber numbers used in the present experiment were 52.1, 57.6, and 63.1. Three kinds of solid surfaces with different wettability were used. These were normal stainless steel (NSS), TiO2 coated NSS, and TiO2 coated NSS radiated with ultraviolet rays. The surface temperatures were varied from 60 to 200 °C. The image of side the view and 30° from horizontal were taken to explain the spreading and the interfacial behavior of a single droplet during impact the hot solid surfaces. It was found that under medium Weber numbers, the surface wettability plays an important role on the droplet spreading and evaporation time during the impact on the hot solid surfaces. The higher the wettability, the larger the droplet spreading on the hot surface, and the lower the evaporation time.

  11. A simplified radiometabolite analysis procedure for PET radioligands using a solid phase extraction with micellar medium

    International Nuclear Information System (INIS)

    Nakao, Ryuji; Halldin, Christer

    2013-01-01

    A solid phase extraction method has been developed for simple and high-speed direct determination of PET radioligands in plasma. Methods: This methodology makes use of a micellar medium and a solid-phase extraction cartridge for displacement of plasma protein bound radioligand and separation of PET radioligands from their radiometabolites without significant preparation. The plasma samples taken from monkey or human during PET measurements were mixed with a micellar eluent containing an anionic surfactant sodium dodecyl sulphate and loaded onto SPE cartridges. The amount of radioactivity corresponding to parent radioligand (retained on the cartridge) and its radioactive metabolites (eluted with micellar eluent) was measured. Results: Under the optimized conditions, excellent separation of target PET radioligands from their radiometabolites was achieved with a single elution and short run-time of 1 min. This method was successfully applied to study the metabolism for 11 C-labelled radioligands in human or monkey plasma. The amount of parent PET radioligands estimated by micellar solid phase extraction strongly corresponded with that determined by radio-LC. The improved throughput permitted the analysis of a large number of plasma samples (up to 13 samples per one PET study) for accurate estimation of metabolite-corrected input function during quantitative PET imaging studies. Conclusion: Solid phase extraction together with micellar medium is fast, sensitive and easy to use, and therefore it is an attractive alternative method to determine relative composition of PET radioligands in plasma

  12. Rat fetal ventral mesencephalon grown as solid tissue cultures

    DEFF Research Database (Denmark)

    Höglinger, G U; Sautter, J; Meyer, Morten

    1998-01-01

    in vitro (DIV) in the presence or absence (controls) of BDNF [100 ng/ml]. The dopamine content in the culture medium, analyzed by HPLC, was significantly higher (4-5 fold) in the BDNF group at DIV 8 and DIV 12 compared to the corresponding control levels (40 pg/ml). The number of tyrosine hydroxylase...

  13. 21 CFR 866.2320 - Differential culture medium.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2320 Differential culture... occurred. Test results aid in the diagnosis of disease and also provide epidemiological information on...

  14. A defined medium for Leishmania culture allows definition of essential amino acids.

    Science.gov (United States)

    Nayak, Archana; Akpunarlieva, Snezhana; Barrett, Michael; Burchmore, Richard

    2018-02-01

    Axenic culture of Leishmania is generally performed in rich, serum-supplemented media which sustain robust growth over multiple passages. The use of such undefined media, however, obscures proteomic analyses and confounds the study of metabolism. We have established a simple, defined culture medium that supports the sustained growth of promastigotes over multiple passages and which yields parasites that have similar infectivity to macrophages to parasites grown in a conventional semi-defined medium. We have exploited this medium to investigate the amino acid requirements of promastigotes in culture and have found that phenylalanine, tryptophan, arginine, leucine, lysine and valine are essential for viability in culture. Most of the 20 proteogenic amino acids promote growth of Leishmania promastigotes, with the exception of alanine, asparagine, and glycine. This defined medium will be useful for further studies of promastigote substrate requirements, and will facilitate future proteomic and metabolomic analyses. Copyright © 2018 Elsevier Inc. All rights reserved.

  15. Study on bystander effect and associated mechanism mediated through culture medium

    International Nuclear Information System (INIS)

    Tu Xumin; Lei Suwen; Zhang Zhixing; Lv Huimin

    2005-01-01

    Objective: To study the bystander effect and associated mechanism mediated through the irradiated cell culture medium. Methods: Splenic natural killer (NK) cells were obtained from healthy male ICR strain mice. Culture medium irradiated with different doses of 60 Co γ-rays was used for culturing Yac-I lymphoma cells. The degree of injury of the latter by activated NK cells was observed. A part of the culture media were pretreated with 1% DMSO, a scavenger of reactive oxygen species (ROS), in order to investigate the possible mechanism of a radiation-induced bystander response. Results: Severer injury was induced in Yac-I cells cultured in the media pre-irradiated with different doses of γ-rays than that in Yac-I cells cultured in unirradiated medium, as shown by increased sensitivity to murine splenic NK cells (P<0.01). Culturing Yac-I cells in DMSO-pretreated medium considerably reduced the activation of NK cells, especially in 0.25 Gy and 0.5 Gy γ-irradiated media. Therefore, it can be expected that DMSO can partly suppress ROS-induced bystander effect. Conclusion: The irradiated culture medium of Yac-I cells can trigger bystander effect. ROS likely plays an important role in radiation-induced bystander effect that can be partly suppressed by pretreatment with DMSO. (authors)

  16. Evaluation of conidia production and mycelial growth in solid culture ...

    African Journals Online (AJOL)

    It is important to know the ability of native strains to carry out the process of sporulation and growth in different conditions and to determine their possible potential as biological control of pests of agricultural importance, mainly in citrus areas. The objective of this study was to evaluate five different solid culture media for the ...

  17. Selection of culture medium and conditions for the production of ...

    African Journals Online (AJOL)

    oyaide

    2013-05-15

    May 15, 2013 ... improving the productivity and economical benefits in livestock production ... was to improve the yeast biomass production measured as dry cell ... the total livestock population in India was 1708 Million ... Media for culture maintenance and optimization .... which is very economical and efficient source for the.

  18. Culturing bovine nucleus pulposus explants by balancing medium osmolarity

    NARCIS (Netherlands)

    Dijk, van B.G.M.; Potier, E.; Ito, K.

    2011-01-01

    Regenerative therapies are promising treatments for early intervertebral disc degeneration. To test their efficacy, an in vitro tissue-level model would be valuable. Nucleus pulposus (NP) explant culture may constitute such a model, as the earliest signs of degeneration are in the NP. However, in NP

  19. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

    Science.gov (United States)

    2013-01-01

    Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into

  20. USE OF SODIUM HIPOCHLORITE IN STERILIZATION OF CULTURE MEDIUM FOR MULTIPLICATION OF Eucalyptus pellita L.

    Directory of Open Access Journals (Sweden)

    Silvio Lopes Teixeira

    2009-10-01

    Full Text Available Lately it has been observed a great interest in the research area of plant tissue culture in discovering new alternatives leading to cost reduction of the plants produced in commercial laboratories, in order to turn this alternative of plant propagation more economical. A potentially promising alternative for this reduction of costs, but which has not been receiving the due attention, is the possibility of substituting the autoclaving technique to a more economical one. With this purpose, two tests were carried out, using a new protocol of medium preparation, which consisted of the chemical sterilization of all the utensils used in the preparation and packaging of the culture medium as well, associated to the addition of the sterilizing agent to the medium, in different concentrations. The objective of the first test was to observe the influence of different concentrations of NaClO added to the culture medium, on its sterilization. The second test aimed at verifying the reaction of the Eucalyptus pellita tissues to different concentrations of NaClO in the culture medium. The addition of NaClO to the culture medium, equal or higher than 0.0005% in the fist test and of 0.005% in the second one, allowed complete sterilization of the medium, without observing any damage to the Eucalyptus pellita tissues, even when they were grown on culture medium containing up to 0.009%, the maximum concentration tried. The results showed the viability of eliminating the autoclave for the sterilization of culture media.

  1. The Stimulatory Effect of Notochordal-Cell Conditioned Medium in a Nucleus Pulposus Explant Culture

    NARCIS (Netherlands)

    de Vries, Stefan; Doeselaar, Marina van; Meij, Björn; Tryfonidou, M; Ito, Keita

    2015-01-01

    OBJECTIVES: Notochordal cell-conditioned medium (NCCM) has previously shown to have a stimulatory effect on nucleus pulposus cells (NPCs) and bone marrow stromal cells (BMSCs) in alginate and pellet cultures. These culture methods provide a different environment than the nucleus pulposus (NP)

  2. The Stimulatory Effect of Notochordal Cell-Conditioned Medium in a Nucleus Pulposus Explant Culture

    NARCIS (Netherlands)

    de Vries, Stefan A H; van Doeselaar, Marina; Meij, Björn P; Tryfonidou, Marianna A; Ito, K

    2016-01-01

    Objectives: Notochordal cell-conditioned medium (NCCM) has previously shown to have a stimulatory effect on nucleus pulposus cells (NPCs) and bone marrow stromal cells (BMSCs) in alginate and pellet cultures. These culture methods provide a different environment than the nucleus pulposus (NP)

  3. Corrosion behavior of HPT-deformed TiNi alloys in cell culture medium

    Science.gov (United States)

    Shri, D. N. Awang; Tsuchiya, K.; Yamamoto, A.

    2017-09-01

    In recent years there are growing interest in fabrication of bulk nanostructured metals and alloys by using severe plastic deformation (SPD) techniques as new alternative in producing bulk nanocrystalline materials. These techniques allows for processing of bulk, fully dense workpiece with ultrafine grains. Metal undergoes SPD processing in certain techniques such as high pressure torsion (HPT), equal-channel angular pressing (ECAP) or multi-directional forging (MDF) are subjected to extensive hydrostatic pressure that may be used to impart a very high strain to the bulk solid without the introduction of any significant change in overall dimension of the sample. The change in the structure (small grain size and high-volume fraction of grain boundaries) of the material may result in the corrosion behavior different from that of the coarse-grained material. Electrochemical measurements were done to understand the corrosion behavior of TiNi alloys before and after HPT deformation. The experiment was carried out using standard three electrode setup (a sample as working electrode; a platinum wire as a counter electrode and a saturated calomel electrode in saturated KCl as a reference electrode) with the surface area of 26.42 mm2 exposed to the EMEM+10% FBS cell culture medium. The measurements were performed in an incubator with controlled environment at 37 °C and 5% CO2, simulating the cell culture condition. The potential of the specimen was monitored over 1 hour, and the stabilized potential was used as the open-circuit potential (EOCP). Potentiodynamic curves were scanned in the potential range from -0.5 V to 1.5 V relative to the EOCP, at a rate of 0.5 mV/s. The result of OCP-time measurement done in the cell culture medium shows that the OCP of HPT-deformed samples shifts towards to the more positive rather than that of BHPT samples. The OCP of deformed samples were ennobled to more than +70 mV for Ti-50mol%. The shift of OCP towards the nobler direction

  4. Ukrainian brown-coal tars recovered at low-temperature carbonization with solid heating medium

    Energy Technology Data Exchange (ETDEWEB)

    Kuznetsov, V I; Govorova, R P; Fadeicheva, A G; Kigel, T B; Chernykh, M K

    1955-01-01

    Three samples of tar were recovered in the laboratory from brown coals carbonized at 375/sup 0/ to 456/sup 0/ +- 25/sup 0/ in a retort with inner heating by solid circulating medium, namely, semicoke (ratio: 4 or 3:1) first heated to 700/sup 0/. One comparative (parallel) experiment was carried out in a retort with inner heating by inert gases entering the retort at 580/sup 0/ to 600/sup 0/ and leaving it at 115/sup 0/ to 120/sup 0/. The tars that were recovered from the retort with the solid heating medium contained a high percentage of coal dust and moisture, which were separated from the tars in supercentrifuges (15,000 rpm). Four samples of cleaned tars were fractionated in a Cu flask with a 2-ball fractional column. The tars from the retort with the solid-heating medium are characterized by increased yield of the petroleum-ether fraction (16.3 or 19.3%) and decreased yield of the paraffin fraction (15.1 to 21.2%) in comparison with those of tar from the retort with gas heating (5.9% of the petroleum ether fraction and 36.5% of paraffin fraction). The yield of paraffin from the paraffin fraction also decreased from 90.6% to 62.6-74.3%. This result shows that in the first case the carbonized products were cracked to a higher degree than those from the retort with gas heating. In raw phenols recovered from fractions of investigated tars, the yield of the phenol-cresol fraction (182/sup 0/ to 204/sup 0/) decreased from 25.9% to 13.0-18.9%.

  5. Ejection of fast recoil atoms from solids under ion bombardment (medium-energy ion scattering by solid surfaces: Pt. 3)

    International Nuclear Information System (INIS)

    Dodonoy, A.I.; Mashkova, E.S.; Molchanov, V.A.

    1989-01-01

    This paper is the third part of our review surface scattering. Part I, which was devoted to the scattering of ions by the surfaces of disordered solids, was published in 1972; Part II, concerning scattering by crystal surfaces, was published in 1974. Since the publication of these reviews the material contained in them has become obsolete in many respects. A more recent account of the status of the problem has been given in a number of studies, including the book by E.S. Mashkova and V.A. Molchanov, Medium-Energy Ion Scattering by Solid Surfaces (Atomizdat, Moscow, 1980), than extended version of which was published by North-Holland in 1985. We note, however, that at the time these reviews were written the study of fast recoil atoms had not been carried out systematically; the problem was studied only as a by-product of surface scattering and sputtering. For this reason, in the above-mentioned works and in other reviews the data relating to recoil atoms were considered only occasionally. In recent years there have appeared a number of works - theoretical, experimental and computer -specially devoted to the study of the ejection of recoil atoms under ion bombardment. A number of interesting effects, which are due to the crystal structure of the target, have been discovered. It therefore, appeared desirable to us to systematize the available material and to present it as Part III of our continuing review. (author)

  6. Proteomic analysis of extracellular proteins from Aspergillus oryzae grown under submerged and solid-state culture conditions.

    Science.gov (United States)

    Oda, Ken; Kakizono, Dararat; Yamada, Osamu; Iefuji, Haruyuki; Akita, Osamu; Iwashita, Kazuhiro

    2006-05-01

    Filamentous fungi are widely used for the production of homologous and heterologous proteins. Recently, there has been increasing interest in Aspergillus oryzae because of its ability to produce heterologous proteins in solid-state culture. To provide an overview of protein secretion by A. oryzae in solid-state culture, we carried out a comparative proteome analysis of extracellular proteins in solid-state and submerged (liquid) cultures. Extracellular proteins prepared from both cultures sequentially from 0 to 40 h were subjected to two-dimensional electrophoresis, and protein spots at 40 h were identified by peptide mass fingerprinting using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. We also attempted to identify cell wall-bound proteins of the submerged culture. We analyzed 85 spots from the solid-state culture and 110 spots from the submerged culture. We identified a total of 29 proteins, which were classified into 4 groups. Group 1 consisted of extracellular proteins specifically produced in the solid-state growth condition, such as glucoamylase B and alanyl dipeptidyl peptidase. Group 2 consisted of extracellular proteins specifically produced in the submerged condition, such as glucoamylase A (GlaA) and xylanase G2 (XynG2). Group 3 consisted of proteins produced in both conditions, such as xylanase G1. Group 4 consisted of proteins that were secreted to the medium in the solid-state growth condition but trapped in the cell wall in the submerged condition, such as alpha-amylase (TAA) and beta-glucosidase (Bgl). A Northern analysis of seven genes from the four groups suggested that the secretion of TAA and Bgl was regulated by trapping these proteins in the cell wall in submerged culture and that secretion of GlaA and XynG2 was regulated at the posttranscriptional level in the solid-state culture.

  7. Dissolved oxygen concentration in the medium during cell culture: Defects and improvements.

    Science.gov (United States)

    Zhang, Kuan; Zhao, Tong; Huang, Xin; He, Yunlin; Zhou, Yanzhao; Wu, Liying; Wu, Kuiwu; Fan, Ming; Zhu, Lingling

    2016-03-01

    In vitro cell culture has provided a useful model to study the effects of oxygen on cellular behavior. However, it remains unknown whether the in vitro operations themselves affect the medium oxygen levels and the living states of cells. In addition, a prevailing controversy is whether reactive oxygen species (ROS) production is induced by continuous hypoxia or reoxygenation. In this study, we have measured the effects of different types of cell culture containers and the oxygen environment where medium replacement takes place on the actual oxygen tension in the medium. We found that the deviations of oxygen concentrations in the medium are much greater in 25-cm(2) flasks than in 24-well plates and 35-mm dishes. The dissolved oxygen concentrations in the medium were increased after medium replacement in normoxia, but remained unchanged in glove boxes in which the oxygen tension remained at a low level (11.4, 5.7, and 0.5% O2 ). We also found that medium replacement in normoxia increased the number of ROS-positive cells and reduced the cell viability; meanwhile, medium replacement in a glove box did not produce the above effects. Therefore, we conclude that the use of 25-cm(2) flasks should be avoided and demonstrate that continuous hypoxia does not produce ROS, whereas the reoxygenation that occurs during the harvesting of cells leads to ROS and induces cell death. © 2015 International Federation for Cell Biology.

  8. High Temperature Gas-to-Gas Heat Exchanger Based on a Solid Intermediate Medium

    Directory of Open Access Journals (Sweden)

    R. Amirante

    2014-04-01

    Full Text Available This paper proposes the design of an innovative high temperature gas-to-gas heat exchanger based on solid particles as intermediate medium, with application in medium and large scale externally fired combined power plants fed by alternative and dirty fuels, such as biomass and coal. An optimization procedure, performed by means of a genetic algorithm combined with computational fluid dynamics (CFD analysis, is employed for the design of the heat exchanger: the goal is the minimization of its size for an assigned heat exchanger efficiency. Two cases, corresponding to efficiencies equal to 80% and 90%, are considered. The scientific and technical difficulties for the realization of the heat exchanger are also faced up; in particular, this work focuses on the development both of a pressurization device, which is needed to move the solid particles within the heat exchanger, and of a pneumatic conveyor, which is required to deliver back the particles from the bottom to the top of the plant in order to realize a continuous operation mode. An analytical approach and a thorough experimental campaign are proposed to analyze the proposed systems and to evaluate the associated energy losses.

  9. STUDIES REGARDING CULTURE MEDIUM INFLUENCE ON IN VITRO REGENERATION FROM WHEAT IMATURE EMBRYOS

    Directory of Open Access Journals (Sweden)

    M. DANCI

    2008-05-01

    Full Text Available Surnamed „embryos’ saving method”, embryos culture is an in vitro technique used for over half of the century for saving the distant hybridization products, that would have degenerate in other conditions. Immature embryos culture is used for initiation of in vitro cultures imposed by the impossibility of using other explants for some of the plant species. Wheat is one of the crops that immature embryos culture technique is suitable for. This methods principle is based on aseptic embryos excision and their inoculation to an adequate culture medium. In vitro culture results depend in a greater manner of the basic culture medium and the hormonal balance used. Immature embryos isolated from two Romanian wheat cultivars – Dropia and Lovrin 41 – were inoculated for callus production on two types of basic media added with 2,4 D. The selected calluses were transferred on MS basic medium and several parameters were registered. Both cultivars emphasized a good callusing capacity, in a different percentage depending on the culture media used, such as 71,09 – 94,45%.. big differences between the cultivars regarding embriogenic callus frequency, shooting callus frequency and regenerated plants percentage were registered.

  10. Uniform, stable supply of medium for in vitro cell culture using a robust chamber

    Science.gov (United States)

    Wei, Juan; Liu, Chong; Jiang, Yang; Liu, Tao; Chen, Li; Liu, Bo; Li, Jingmin

    2018-06-01

    A uniform, stable supply of medium is important for in vitro cell culture. In this paper, a microfluidic device is presented for culturing cells inside a robust chamber with continuous perfusion of medium. The device consists of a main channel, two bifurcated channels and a culture chamber. The culture chamber connects to the bifurcated channels via multiple paths, and distributes symmetrically on the main channel, to improve the efficiency of medium exchange. Furthermore, regular polygonal chambers with various numbers of edges have been designed, to study the effects of chamber shape on flow fields. The finite element method has been employed to predict the effects of multiple paths on the uniformity and stability of flow fields in the culture chamber. Particle tracking technology has been used to evaluate the flow fields in the chambers, and PC-12 cells have been cultured using the microfluidic device, to test its validity. The results of simulation and experiment indicate that the microfluidic design could provide a continuous interstitial-like flow microenvironment, with a relatively stable and uniform supply of medium.

  11. Effect of the reaction medium on the properties of solid catalysts

    Energy Technology Data Exchange (ETDEWEB)

    Boreskov, G.K.

    1980-01-01

    The effect of the reaction medium on the properties of solid catalysts, such as bulk or supported metals, alloys, or metal oxides, include variations in surface composition, structure, and catalytic properties due to catalyst interaction with the reactants. This interaction leads to the establishment of a steady state, which is determined by the composition of the reaction medium and temperature, but is independent of the initial state of the catalyst. This steady state for a catalyst of a given chemical composition is characterized by an approximately constant specific activity in most chemical reactions, which is almost independent of the preparation method, surface area, or crystal size of the catalyst. The structurally sensitive reactions, which occur only on limited segments of catalyst surface characterized by specific structures, are the exception. The effects of the variations in catalytic properties caused by the reaction medium on the steady-state and nonsteady-state reaction kinetics are also discussed based on the results obtained for oxidative dehydrogenation of 1-butene over an iron/antimony oxide catalyst.

  12. Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium.

    Science.gov (United States)

    Xella, Susanna; Marsella, Tiziana; Tagliasacchi, Daniela; Giulini, Simone; La Marca, Antonio; Tirelli, Alessandra; Volpe, Annibale

    2010-04-01

    To compare the outcome of two different culture media marketed by the MediCult AS Company (Jyllinge, Denmark)-Universal IVF Medium and ISM1 Medium culture-which, in addition to glucose, pyruvate, and energy-providing components, also contain amino acids, nucleotides, vitamins, and cholesterol. Laboratory and retrospective clinical study. University teaching hospital. A total of 726 patients, undergoing IVF-intracytoplasmic sperm injection procedure, comparable in mean age range, oocyte retrieval, and infertility indication, were included in the study. Laboratory quality and standard procedures were maintained unaffected. Oocyte retrieval, different embryo culture media. Embryo quality, ongoing pregnancy, and implantation rate. The frequency of good-quality embryos (79% vs. 74%) and the percentages of ongoing pregnancy (27.5% vs. 18%) and implantation rate (15% vs. 10%) were significantly higher in the group treated with ISM1 Medium rather than Universal IVF Medium. ISM1 Medium culture seems to improve the performance of embryonic growth and development, as well as increasing the percentage of pregnancy. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. Utilization and optimization of a waste stream cellulose culture medium for pigment production by Penicillium spp.

    Science.gov (United States)

    Sopandi, T; Wardah, A; Surtiningsih, T; Suwandi, A; Smith, J J

    2013-03-01

    This research sought to determine optimal corn waste stream-based fermentation medium C and N sources and incubation time to maximize pigment production by an indigenous Indonesian Penicillium spp., as well as to assess pigment pH stability. A Penicillium spp. was isolated from Indonesian soil, identified as Penicillium resticulosum, and used to test the effects of carbon and nitrogen type and concentrations, medium pH, incubation period and furfural on biomass and pigment yield (PY) in a waste corncob hydrolysate basal medium. Maximum red PY (497.03 ± 55.13 mg l(-1)) was obtained with a 21 : 1 C : N ratio, pH 5.5-6.0; yeast extract-, NH(4) NO(3)-, NaNO(3)-, MgSO(4) ·7H(2) O-, xylose- or carboxymethylcellulose (CMC)-supplemented medium and 12 days (25 °C, 60-70% relative humidity, dark) incubation. C source, C, N and furfural concentration, medium pH and incubation period all influenced biomass and PY. Pigment was pH 2-9 stable. Penicillium resticulosum demonstrated microbial pH-stable-pigment production potential using a xylose or CMC and N source, supplemented waste stream cellulose culture medium. Corn derived, waste stream cellulose can be used as a culture medium for fungal pigment production. Such application provides a process for agricultural waste stream resource reuse for production of compounds in increasing demand. © 2012 The Society for Applied Microbiology.

  14. The natural selection of organizational and safety culture within a small to medium sized enterprise (SME).

    Science.gov (United States)

    Brooks, Benjamin

    2008-01-01

    Small to Medium Sized Enterprises (SMEs) form the majority of Australian businesses. This study uses ethnographic research methods to describe the organizational culture of a small furniture-manufacturing business in southern Australia. Results show a range of cultural assumptions variously 'embedded' within the enterprise. In line with memetics - Richard Dawkin's cultural application of Charles Darwin's theory of Evolution by Natural Selection, the author suggests that these assumptions compete to be replicated and retained within the organization. The author suggests that dominant assumptions are naturally selected, and that the selection can be better understood by considering the cultural assumptions in reference to Darwin's original principles and Frederik Barth's anthropological framework of knowledge. The results are discussed with reference to safety systems, negative cultural elements called Cultural Safety Viruses, and how our understanding of this particular organizational culture might be used to build resistance to these viruses.

  15. Optimizing in vitro large scale production of giant reed (Arundo donax L.) by liquid medium culture

    International Nuclear Information System (INIS)

    Cavallaro, Valeria; Patanè, Cristina; Cosentino, Salvatore L.; Di Silvestro, Isabella; Copani, Venera

    2014-01-01

    Tissue culture methods offer the potential for large-scale propagation of giant reed (Arundo donax L.), a promising crop for energy biomass. In previous trials, giant reed resulted particularly suitable to in vitro culture. In this paper, with the final goal of enhancing the efficiency of in vitro production process and reducing costs, the influence of four different culture media (agar or gellan-gum solidified medium, liquid medium into a temporary immersion system-RITA ® or in a stationary state) on in vitro shoot proliferation of giant reed was evaluated. Giant reed exhibited a particular sensitivity to gelling agents during the phase of secondary shoot formation. Gellan gum, as compared to agar, improved the efficiency of in vitro culture giving more shoots with higher mean fresh and dry weight. Moreover, the cultivation of this species into a liquid medium under temporary immersion conditions or in a stationary state, was comparatively as effective as and cheaper than that into a gellan gum medium. Increasing 6-benzylaminopurine (BA) up to 4 mg l −1 also resulted in a further enhancement of secondary shoot proliferation. The good adaptability of this species to liquid medium and the high multiplication rates observed indicate the possibility to obtain from a single node at least 1200 plantlets every six multiplication cycles (about 6 months), a number 100 fold higher than that obtained yearly per plant by the conventional methods of vegetative multiplication. In open field, micropropagated plantlets guaranteed a higher number of survived plants, secondary stems and above ground biomass as compared to rhizome ones. - Highlights: • In vitro propagation offers the potential for large-scale propagation of giant reed. • The success of an in vitro protocol depends on the rate and mode of shoot proliferation. • Substituting liquid media to solid ones may decrease propagation costs in Arundo donax. • Giant reed showed good proliferation rates in

  16. The release of elements from dental casting alloy into cell-culture medium and artificial saliva.

    Science.gov (United States)

    Can, Gülşen; Akpınar, Gül; Aydın, Ahmet

    2007-04-01

    The biocompatibility of dental casting alloys is a critical issue because these alloys are in long-term intimate contact with oral tissues. Since the biocompatibility of alloys is not completely known; the release of elements from the alloys has been studied. The aim of this study was to compare the elemental release from dental casting alloy during exposure to artificial saliva and cell-culture medium. Twenty specimens made from Ni-Cr alloy were provided in the form of 5 mm diameter discs, 2 mm in thickness with a 7 mm stem attached to one face to facilitate handling. Ten of twenty samples were polished separately using a conventional technique. The remaining ten samples were left sandblasted with 50 mum Al(2)0(3). Ten samples (5 polished, 5 sandblasted) were separately placed into cell-culture wells with Dulbecco's Modified Eagle's Medium. The other ten samples were placed separately into cell-culture wells with artificial saliva. The samples were subjected in contact with these medium for 30 days. These medium were collected every 7 days. The cell-culture medium and artificial saliva without alloy samples were subjected to elemental analyses as a control. At the end of the exposure time, Atomic Absorption Spectrometry (AAS) was used to determine the release of elements from the alloys into all collected medium. Statistical analyses were assessed with two-way ANOVA. In general, the elemental release occurred with in all medium. The elemental releases of sandblasted alloys were higher than polished alloys. Artificial saliva was found to cause more release from the samples. In both media, Ni released from polished and sandblasted alloys were higher than Cr and Mo. The results suggest that the release of elements from the alloys might have correlated with the environments and the surface of dental alloy.

  17. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

    Directory of Open Access Journals (Sweden)

    Céline Bouillon

    Full Text Available In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371 were randomized according to two media (Single Step Medium--SSM group or Global medium (Global group. This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73 conceived in the randomized study were included (42 for Global and 31 for SSM. The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded. The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05. The culture medium had no significant effect on birthweight, risk of malformation (minor and major, growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002, irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

  18. The type of culture medium and the duration of in vitro culture do not influence birthweight of ART singletons.

    Science.gov (United States)

    De Vos, A; Janssens, R; Van de Velde, H; Haentjens, P; Bonduelle, M; Tournaye, H; Verheyen, G

    2015-01-01

    Does the type of in vitro culture medium or the duration of in vitro culture influence singleton birthweight after IVF/ICSI treatment? In a comparison of two culture media, neither the medium nor the duration of culture (Day 3 versus Day 5 blastocyst transfer) had any effect on mean singleton birthweight. Previous studies indicated that in vitro culture of human embryos may affect birthweight of live born singletons. Both the type of culture medium and the duration of culture may be implicated. However, these studies are small and report conflicting results. A large retrospective analysis was performed including all singleton live births after transferring fresh Day 3 or Day 5 embryos. IVF and ICSI cycles performed between April 2004 and December 2009 at a tertiary care centre were included for analysis. A total of 2098 singleton live births resulting from singleton pregnancies were included for analysis. Two different sequential embryo culture media were concurrently used in an alternating way: Medicult (n = 1388) and Vitrolife (n = 710). Maternal age, maternal and paternal BMI, maternal parity, maternal smoking, main cause of infertility, cycle rank, stimulation protocol, method of fertilization (IVF or ICSI), time in culture and number of embryos transferred were taken into account. Embryo transfers were performed either on Day 3 (n = 1234) or on Day 5 (n = 864). Singleton birthweight was the primary outcome parameter. Gestational age and gender of the newborn were accounted for in the multiple regression analysis. No significant differences in mean singleton birthweight were observed between the two culture media: Medicult 3222 g (±15 SE) and Vitrolife 3251 g (±21 SE) (P = 0.264). The mean singleton birthweight was not different between Day 3 embryo transfers (3219 ± 16 g) and Day 5 blastocyst transfers (3250 ± 19 g; P = 0.209). Multiple regression analysis controlling for potential maternal, paternal, treatment and newborn confounders confirmed the non

  19. Selection of osteoprogenitors from the jaw periosteum by a specific animal-free culture medium.

    Directory of Open Access Journals (Sweden)

    Dorothea Alexander

    Full Text Available The goal of our research work is to establish mesenchymal osteoprogenitors derived from human jaw periosteum for tissue engineering applications in oral and maxillofacial surgery. For future autologous and/or allogeneic transplantations, some issues must be addressed. On the one hand, animal-free culture conditions have yet to be established. On the other hand, attempts should be undertaken to shorten the in vitro culturing process efficiently. The aim of the present study is to compare and analyze the phenotype of osteoprogenitors from the jaw periosteum under normal FCS-containing and animal-free culture conditions. Therefore, we analyzed the proliferation rates of MesenCult-XF medium (MC- in comparison to DMEM-cultured JPCs. Whereas jaw periosteal cells (JPCs show relatively slow proliferation rates and a fibroblastoid shape under DMEM culture conditions, MC-cultured JPCs diminished their cell size significantly and proliferated rapidly. By live-monitoring measurements of adhesion and proliferation, we made an interesting observation: whereas the proliferation of the MSCA-1(+ subpopulation and the unseparated cell fraction were favored by the animal-free culture medium, the proliferation of the MSCA-1(- subpopulation seemed to be repressed under these conditions. The alkaline phosphatase expression pattern showed similar results under both culture conditions. Comparison of the mineralization capacity revealed an earlier formation of calcium-phosphate precipitates under MC culture conditions; however, the mineralization capacity of the DMEM-cultured cells seemed to be higher. We conclude that the tested animal-free medium is suitable for the in vitro expansion and even for the specific selection of osteoprogenitor cells derived from the jaw periosteum.

  20. Sensitive and selective culture medium for detection of environmental Clostridium difficile isolates without requirement for anaerobic culture conditions.

    Science.gov (United States)

    Cadnum, Jennifer L; Hurless, Kelly N; Deshpande, Abhishek; Nerandzic, Michelle M; Kundrapu, Sirisha; Donskey, Curtis J

    2014-09-01

    Effective and easy-to-use methods for detecting Clostridium difficile spore contamination would be useful for identifying environmental reservoirs and monitoring the effectiveness of room disinfection. Culture-based detection methods are sensitive for detecting C. difficile, but their utility is limited due to the requirement of anaerobic culture conditions and microbiological expertise. We developed a low-cost selective broth medium containing thioglycolic acid and l-cystine, termed C. difficile brucella broth with thioglycolic acid and l-cystine (CDBB-TC), for the detection of C. difficile from environmental specimens under aerobic culture conditions. The sensitivity and specificity of CDBB-TC (under aerobic culture conditions) were compared to those of CDBB (under anaerobic culture conditions) for the recovery of C. difficile from swabs collected from hospital room surfaces. CDBB-TC was significantly more sensitive than CDBB for recovering environmental C. difficile (36/41 [88%] versus 21/41 [51%], respectively; P = 0.006). C. difficile latex agglutination, an enzyme immunoassay for toxins A and B or glutamate dehydrogenase, and a PCR for toxin B genes were all effective as confirmatory tests. For 477 total environmental cultures, the specificity of CDBB-TC versus that of CDBB based upon false-positive yellow-color development of the medium without recovery of C. difficile was 100% (0 false-positive results) versus 96% (18 false-positive results), respectively. False-positive cultures for CDBB were attributable to the growth of anaerobic non-C. difficile organisms that did not grow in CDBB-TC. Our results suggest that CDBB-TC provides a sensitive and selective medium for the recovery of C. difficile organisms from environmental samples, without the need for anaerobic culture conditions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  1. Effect of Cold Stress on Fruiting Body Production by Medicinal Basidiomycetes in Submerged and Solid-phase Culture

    Directory of Open Access Journals (Sweden)

    E.P. Vetchinkina

    2017-05-01

    Full Text Available The ability of the medicinal xylotrophic basidiomycetes Lentinus edodes, Pleurotus ostreatus, Ganoderma lucidum and Grifola frondosa to produce typical and atypical fruiting bodies with viable basidiospores in submerged and solid-phase culture under stationary conditions in a beer wort-containing medium under cold stress was shown. The examined mushrooms, when not exposed to temperature stress, did not form fruiting bodies. In solid-phase culture in an agarized medium after cold treatment, the basidiome formation period was shortened by 1.5–2 times. Furthermore, the use of a mycelium subjected to temperature stress for inoculation induced and accelerated the formation of fruiting bodies on an industrial wood substrate, which is of great biotechnological importance.

  2. Emerging Culture of English-Medium Instruction in Korea: Experiences of Korean and International Students

    Science.gov (United States)

    Kim, Jeongyeon; Tatar, Bradley; Choi, Jinsook

    2014-01-01

    This study aims to contrastively examine Korean and international students' experiences of taking subject courses at a Korean university. Focusing on the viewpoints of the students, rather than central authorities, we attempt to reveal how language use and cultural factors are interpenetrated in the praxis of English-medium instruction (EMI). The…

  3. 21 CFR 866.2410 - Culture medium for pathogenic Neisseria spp.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Culture medium for pathogenic Neisseria spp. 866.2410 Section 866.2410 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2410...

  4. A procedure for culturing rat neocortex explants in a serum-free nutrient medium

    NARCIS (Netherlands)

    Romijn, H. J.; de Jong, B. M.; Ruijter, J. M.

    1988-01-01

    A procedure is described for long-term culturing of rat neocortex explants in a serum-free growth medium. Slices spanning the entire cortical depth from pial to ventricular side are prepared from 6-day-old rat pups. After preincubation in Hanks' balanced salt solution with extra glucose, the

  5. Studies on irradiated BNFL culture medium for decontamination and longer storage

    International Nuclear Information System (INIS)

    Singh, Antaryami; Malodia, P.; Jain, S.K.; Ram Gopal

    2001-01-01

    The feasibility of gamma radiation for microbial decontamination and shelf-life extension of culture medium was studied. Changes in total viable count, coliform count and fungal count on exposure to 5, 10, 15, 20 and 25 kGy of gamma radiation were examined. The total viable counts were reduced on irradiation. Complete destruction of bacterial and fungal contamination was observed at 20 kGy. Studies were conducted to examine the changes in microbial contamination of the medium during storage. There was no post irradiation proliferation of microorganisms. Also, no significant change in the efficiency of the irradiated culture medium was observed. Thus, irradiation is extremely useful for longer storage and quality-assurance. (author)

  6. Calcium Concentration in Culture Medium as a Nondestructive and Rapid Marker of Osteogenesis.

    Science.gov (United States)

    Tanikake, Yohei; Akahane, Manabu; Furukawa, Akira; Tohma, Yasuaki; Inagaki, Yusuke; Kira, Tsutomu; Tanaka, Yasuhito

    2017-06-09

    Artificial bones made of β-tricalcium phosphate (β-TCP) combined with bone marrow-derived mesenchymal stromal cells (BM-MSCs) are used for effective reconstruction of bone defects caused by genetic defects, traumatic injury, or surgical resection of bone tumors. However, the selection of constructs with high osteogenic potential before implantation is challenging. The purpose of this study was to determine whether the calcium concentration in BM-MSC culture medium can be used as a nondestructive and simple osteogenic marker for selecting tissue-engineered grafts constructed using β-TCP and BM-MSCs. We prepared three cell passages of BM-MSCs derived from three 7-week-old, male Fischer 344 rats; the cells were cultured in osteoinductive medium in the presence of β-TCP for 15 days. The medium was replaced with fresh medium on day 1 in culture and subsequently changed every 48 h; it was collected for measurement of osteocalcin secretion and calcium concentration by enzyme-linked immunosorbent assay and X-ray fluorescence spectrometry, respectively. After cultivation, the constructs were implanted subcutaneously into the backs of recipient rats. Four weeks after implantation, the alkaline phosphatase (ALP) activity and osteocalcin content of the constructs were measured. A strong inverse correlation was observed between the calcium concentration in the medium and the ALP activity and osteocalcin content of the constructs, with Pearson's correlation coefficients of 0.92 and 0.90, respectively. These results indicate that tissue-engineered bone with high osteogenic ability can be selected before implantation based on low calcium content of the culture medium, resulting in successful bone formation after implantation. This nondestructive, simple method shows great promise for assessing the osteogenic ability of tissue-engineered bone.

  7. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

    Science.gov (United States)

    Bouillon, Céline; Léandri, Roger; Desch, Laurent; Ernst, Alexandra; Bruno, Céline; Cerf, Charline; Chiron, Alexandra; Souchay, Céline; Burguet, Antoine; Jimenez, Clément; Sagot, Paul; Fauque, Patricia

    2016-01-01

    In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium--SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (pculture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

  8. Growth and Development of Colletotrichum gloeosporioides f. alatae During Culture in Liquid Medium

    Directory of Open Access Journals (Sweden)

    Laura E. Cerón Rincón

    2006-01-01

    Full Text Available Some characteristics known as virulence factors for Colletotrichum sp. genus, like: weight of the produced mycelium, sporulation, poligalacturonase activity and pH medium were evaluated during the growth of C. gloeosporioides f. alatae in three liquid medium commonly used for fungi culture (Czapeck, Martin broth and potato broth and additionally (Czapeck with yam extract as the only source of carbon. After of 17 days of growth, maximum values were obtained for the above parameters in the last medium, compared with others growth media evaluated. The implemented medium with yam extract, supply nutritional requirements of the pathogen for the development of characteristic factors related with mechanism of infections that may play a role in the pathogenesis.

  9. Isolation of previously uncultured rumen bacteria by dilution to extinction using a new liquid culture medium.

    Science.gov (United States)

    Kenters, Nikki; Henderson, Gemma; Jeyanathan, Jeyamalar; Kittelmann, Sandra; Janssen, Peter H

    2011-01-01

    A new anaerobic medium that mimics the salts composition of rumen fluid was used in conjunction with a dilution method of liquid culture to isolate fermentative bacteria from the rumen of a grass-fed sheep. The aim was to inoculate a large number of culture tubes each with a mean of 97% sequence identity to genes of uncultured bacteria detected in various gastrointestinal environments. This strategy has therefore allowed us to cultivate many novel rumen bacteria, opening the way to overcoming the lack of cultures of many of the groups detected using cultivation-independent methods. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Culture medium and growth regulators on in vitro multiplication of Psidium guajava L.

    Directory of Open Access Journals (Sweden)

    Jorge Vilchez

    2014-01-01

    Full Text Available Guava (Psidium guajava L. cultivar `Dwarf Cuban Red 18-40 EEA' has high yields. For large-scale propagation, micropropagation is a possible solution. The aim of this study was to determine the effect of two culture media, two cytokinins and an analog brasinoesteoides (DI-31 in the in vitro multiplication of this cultivar. Two culture media (MS and WPM, three concentrations of benzylaminopurine (BAP (0.5, 1.0, 1.5 mg l-1, three of kinetin (0.5, 1.0, 1.5 mg l-1 and two DI-31 (0.01 and 0.02 mg l-1 were evaluated. The variables evaluated were: number of shoots, number of leaves, shoot length and multiplication coefficient. It was found that the type of culture medium influenced the shoot multiplication of guava. The number of shoots, shoot length and multiplication coefficient were determined by the type and concentration of cytokinin added to the culture medium. With the use of WPM culture medium with 1 mg l-1 BAP It was obtained the highest values of the variables evaluated. The use of DI-31 promoted the shoot growth without affecting the multiplication coefficient. Key words: benzylaminopurine, DI-31, kinetin, guava, micropropagation, multiplication phase

  11. Addition of Carbon to the Culture Medium Improves the Detection Efficiency of Aflatoxin Synthetic Fungi

    Directory of Open Access Journals (Sweden)

    Tadahiro Suzuki

    2016-11-01

    Full Text Available Aflatoxin (AF is a harmful secondary metabolite that is synthesized by the Aspergillus species. Although AF detection techniques have been developed, techniques for detection of AF synthetic fungi are still required. Techniques such as plate culture methods are continually being modified for this purpose. However, plate culture methods require refinement because they suffer from several issues. In this study, activated charcoal powder (carbon was added to a culture medium containing cyclodextrin (CD to enhance the contrast of fluorescence and improve the detection efficiency for AF synthetic fungi. Two culture media, potato dextrose agar and yeast extract sucrose agar, were investigated using both plate and liquid cultures. The final concentrations of CD and carbon in the media were 3 mg/mL and 0.3 mg/mL, respectively. Addition of carbon improved the visibility of fluorescence by attenuating approximately 30% of light scattering. Several fungi that could not be detected with only CD in the medium were detected with carbon addition. The carbon also facilitated fungal growth in the potato dextrose liquid medium. The results suggest that addition of carbon to media can enhance the observation of AF-derived fluorescence.

  12. Dependence of the cytotoxicity of multi-walled carbon nanotubes on the culture medium

    International Nuclear Information System (INIS)

    Zhu Ying; Ran Tiecheng; Li Yuguo; Guo Jinxue; Li Wenxin

    2006-01-01

    This study examined the influence of multi-walled carbon nanotubes (MWNTs) on the growth of the unicellular protozoan Tetrahymena pyriformis. Contrary to the findings from most other investigations, our experiment indicated that MWNTs stimulated growth of the cells cultured in proteose peptone yeast extract medium (PPY). Atomic force microscopy images and thermogravimetric analysis showed the spontaneous formation of peptone-MWNT conjugates in the medium by noncovalent binding. Uptake of large amounts of the conjugates by Tetrahymena pyriformis was responsible for growth stimulation, evidenced by images with fluorescently labelled peptone. After the PPY medium was replaced by a filtrated pond water medium (FPW), however, inhibition of the growth of cells exposed to MWNTs occurred. Measurements of the level of malondialdehyde and superoxide dismutase activity demonstrated further that MWNTs might be either toxic or nontoxic, depending on the medium used to cultivate Tetrahymena pyriformis. The biological effects of the interaction of MWNTs with some composites in culture media would be helpful for understanding the mechanisms of the toxicity of carbon nanotubes to living systems

  13. Participation of cob tissue in the transport of medium components into maize kernels cultured in vitro

    International Nuclear Information System (INIS)

    Felker, F.C.

    1990-01-01

    Maize (Zea mays L.) kernels cultured in vitro while still attached to cob pieces have been used as a model system to study the physiology of kernel development. In this study, the role of the cob tissue in uptake of medium components into kernels was examined. Cob tissue was essential for in vitro kernel growth, and better growth occurred with larger cob/kernel ratios. A symplastically transported fluorescent dye readily permeated the endosperm when supplied in the medium, while an apoplastic dye did not. Slicing the cob tissue to disrupt vascular connections, but not apoplastic continuity, greatly reduced [ 14 C]sucrose uptake into kernels. [ 14 C]Sucrose uptake by cob and kernel tissue was reduced 31% and 68%, respectively, by 5 mM PCMBS. L-[ 14 C]glucose was absorbed much more slowly than D-[ 14 C]glucose. These and other results indicate that phloem loading of sugars occurs in the cob tissue. Passage of medium components through the symplast cob tissue may be a prerequisite for uptake into the kernel. Simple diffusion from the medium to the kernels is unlikely. Therefore, the ability of substances to be transported into cob tissue cells should be considered in formulating culture medium

  14. Variation of Spirulina maxima biomass production in different depths of urea-used culture medium.

    Science.gov (United States)

    Affan, Md-Abu; Lee, Dae-Won; Al-Harbi, Salim Marzoog; Kim, Han-Jun; Abdulwassi, Najah Ibrahim; Heo, Soo-Jin; Oh, Chulhong; Park, Heung-Sik; Ma, Chae Woo; Lee, Hyeon-Yong; Kang, Do-Hyung

    2015-01-01

    Fewer studies have assessed the outdoor cultivation of Spirulina maxima compared with S. platensis, although the protein content of S. maxima is higher than S. platensis. Spirulina growth medium requires an increased amount of NaHCO3, Na2CO3, and NaNO3, which increases the production cost. Therefore, the current study used a low-cost but high-efficiency biomass production medium (Medium M-19) after testing 33 different media. The medium depth of 25 cm (group A) was sub-divided into A1 (50% cover with a black curtain (PolyMax, 12 oz ultra-blackout), A2 (25% cover), and A3 (no cover). Similarly the medium depths of 30 and 35 cm were categorized as groups B (B1, B2, and B3) and C (C1, C2, and C3), respectively, and the effects of depth and surface light availability on growth and biomass production were assessed. The highest biomass production was 2.05 g L-1 in group A2, which was significantly higher (p maxima died in B1 and C1 on the fifth day of culture. The biochemical composition of the biomass obtained from A2 cultures, including protein, carbohydrate, lipid, moisture, and ash, was 56.59%, 14.42%, 0.94%, 5.03%, and 23.02%, respectively. Therefore, S. maxima could be grown outdoors with the highest efficiency in urea-enriched medium at a 25-cm medium depth with 25% surface cover or uncovered.

  15. Atrazine and its metabolites degradation in mineral salts medium and soil using an enrichment culture.

    Science.gov (United States)

    Kumar, Anup; Singh, Neera

    2016-03-01

    An atrazine-degrading enrichment culture was used to study degradation of atrazine metabolites viz. hydroxyatrazine, deethylatrazine, and deisopropylatrazine in mineral salts medium. Results suggested that the enrichment culture was able to degrade only hydroxyatrazine, and it was used as the sole source of carbon and nitrogen. Hydroxyatrazine degradation slowed down when sucrose and/or ammonium hydrogen phosphate were supplemented as the additional sources of carbon and nitrogen, respectively. The enrichment culture could degrade high concentrations of atrazine (up to 110 μg/mL) in mineral salts medium, and neutral pH was optimum for atrazine degradation. Further, except in an acidic soil, enrichment culture was able to degrade atrazine in three soil types having different physico-chemical properties. Raising the pH of acidic soil to neutral or alkaline enabled the enrichment culture to degrade atrazine suggesting that acidic pH inhibited atrazine-degrading ability. The study suggested that the enrichment culture can be successfully utilized to achieve complete degradation of atrazine and its persistent metabolite hydroxyatrazine in the contaminated soil and water.

  16. Optimizing a culture medium for biomass and phenolic compounds production using Ganoderma lucidum

    Directory of Open Access Journals (Sweden)

    Carlos Andrés Zárate-Chaves

    2013-01-01

    Full Text Available The present work was aimed at optimizing a culture medium for biomass production and phenolic compounds by using Ganoderma lucidum. The culture was optimized in two stages; a Plackett-Burman design was used in the first one for identifying key components in the medium and a central composite design was used in the second one for optimizing their concentration. Both responses (biomass and phenolic compounds were simultaneously optimized by the latter methodology regarding desirability, and the optimal concentrations obtained were 50.00 g/L sucrose, 13.29 g/L yeast extract and 2.99 g/L olive oil. Maximum biomass production identified in these optimal conditions was 9.5 g/L and that for phenolic compounds was 0.0452 g/L, this being 100% better than that obtained in the media usually used in the laboratory. Similar patterns regarding chemical characterization and biological activity towards Aspergillus sp., from both fruiting body and mycelium-derived secondary metabolites and extracts obtained in the proposed medium were observed. It was shown that such statistical methodologies are useful for optimizing fermentation and, in the specific case of G. lucidum, optimizing processes for its production and its metabolites in submerged culture as an alternative to traditional culture.

  17. In vitro mouse spermatogenesis with an organ culture method in chemically defined medium.

    Directory of Open Access Journals (Sweden)

    Hiroyuki Sanjo

    Full Text Available We previously reported the successful induction and completion of mouse spermatogenesis by culturing neonatal testis tissues. The culture medium consisted of α-minimum essential medium (α-MEM, supplemented with Knockout serum replacement (KSR or AlbuMAX, neither of which were defined chemically. In this study, we formulated a chemically defined medium (CDM that can induce mouse spermatogenesis under organ culture conditions. It was found that bovine serum albumin (BSA purified through three different procedures had different effects on spermatogenesis. We also confirmed that retinoic acid (RA played crucial roles in the onset of spermatogonial differentiation and meiotic initiation. The added lipids exhibited weak promoting effects on spermatogenesis. Lastly, luteinizing hormone (LH, follicle stimulating hormone (FSH, triiodothyronine (T3, and testosterone (T combined together promoted spermatogenesis until round spermatid production. The CDM, however, was not able to produce elongated spermatids. It was also unable to induce spermatogenesis from the very early neonatal period, before 2 days postpartum, leaving certain factors necessary for spermatogenic induction in mice unidentified. Nonetheless, the present study provided important basic information on testis organ culture and spermatogenesis in vitro.

  18. Physiological Response of In Vitro Cultured MAGNOLIA SP. to Nutrient Medium Composition

    Directory of Open Access Journals (Sweden)

    S. Sokolov Rossen

    2014-09-01

    Full Text Available The objective of this study was to assess the regeneration response of in vitro cultured Magnolia × soulangeana ‘Alexandrina’ and Magnolia liliiflora ‘Nigra’ to nutrient medium composition. In the primary culture (initiated from dormant axillary buds combinations of Murashige and Skoog (MS basal salts with 6-benzylaminopurine and α-naphthaleneacetic acid were tested. The primary explants of cv. ‘Alexandrina’ expressed higher regeneration rate than cv. ‘Nigra’. For both species, the regen eration was most strongly potentiated at addition of 0.25 mg dm−3 of the cytokinin alone. The auxin exerted undesir–able effects. Several basal salts media were applied in proliferation stage and their physiological effects were evaluated in reference to traditionally used MS. At culturing on Chée & Pool C2d Vitis Medium (VM that is for the first time introduced to magnolia and on MS, M. liliiflora formed more but less elongated shoots than M. soulangeana. However, on VM, substantial increase (25-30% of the number of axillary shoots and leaves, shoot length and fresh and dry weights over MS was established for both species. This suggested VM as promising composition of nutrients in multiplication stage. Microshoots obtained on MS, VM, Rugini Olive Medium and DKW Juglans Medium were successfully rooted in vitro and subsequently established ex vitro. The findings expand the information on magnolia response to culture conditions and contribute to elaboration of innovative elements of protocols for establishing tissue cultures with high regeneration capacity.

  19. IVF culture medium affects post-natal weight in humans during the first 2 years of life

    NARCIS (Netherlands)

    Kleijkers, Sander H. M.; van Montfoort, Aafke P. A.; Smits, Luc J. M.; Viechtbauer, Wolfgang; Roseboom, Tessa J.; Nelissen, Ewka C. M.; Coonen, Edith; Derhaag, Josien G.; Bastings, Lobke; Schreurs, Inge E. L.; Evers, Johannes L. H.; Dumoulin, John C. M.

    2014-01-01

    Is post-natal growth during the first 2 years of life in IVF singletons affected by type of medium used for culturing human embryos during an IVF treatment? The in vitro culture of human embryos in medium from Cook resulted in singletons with a lower weight during the first 2 years of life compared

  20. Mitochondrial DNA content in embryo culture medium is significantly associated with human embryo fragmentation.

    Science.gov (United States)

    Stigliani, S; Anserini, P; Venturini, P L; Scaruffi, P

    2013-10-01

    Is the amount of cell-free DNA released by human embryos into culture medium correlated with embryo morphological features? The mitochondrial DNA (mtDNA) content of culture medium is significantly associated with the fragmentation rate on Days 2 and 3 of embryo development, whether the oocyte came from women ≤ 35 or >35 years old. Cellular fragmentation is often utilized as one of the morphological parameters for embryo quality assessment. The amount of cellular fragments is considered to be an important morphological parameter for embryo implantation potential. It has been hypothesized that fragments are apoptotic bodies or anuclear cytoplasmatic pieces of blastomeres, although no definitive conclusion has been drawn about their pathogenesis. Human fertilized oocytes were individually cultured from Day 1 to Days 2 and 3. A total of 800 samples (166 spent media from Day 2 and 634 from Day 3) were enrolled into the present study. Double-stranded DNA (dsDNA) was quantified in 800 spent embryo culture media by Pico Green dye fluorescence assay. After DNA purification, genomic DNA (gDNA) and mtDNA were profiled by specific quantitative PCR. Statistical analyses defined correlations among DNA contents, embryo morphology and maternal age. Different independent tests confirmed the presence of DNA into embryo culture medium and, for the first time, we demonstrate that both gDNA and mtDNA are detectable in the secretome. The amount of DNA is larger in embryos with bad quality cleavage compared with high-grade embryos, suggesting that the DNA profile of culture medium is an objective marker for embryo quality assessment. In particular, DNA profiles are significantly associated with fragmentation feature (total dsDNA: P = 0.0010; mtDNA; P = 0.0247) and advanced maternal age. It is necessary to establish whether DNA profiling of spent embryo culture medium is a robust onsite test that can improve the prediction of blastulation, implantation and/or pregnancy rate. The

  1. Plasma generated in culture medium induces damages of HeLa cells due to flow phenomena

    Science.gov (United States)

    Sato, Yusuke; Sato, Takehiko; Yoshino, Daisuke

    2018-03-01

    Plasma in a liquid has been anticipated as an effective tool for medical applications, however, few reports have described cellular responses to plasma generated in a liquid similar to biological fluids. Herein we report the effects of plasma generated in a culture medium on HeLa cells. The plasma in the culture medium produced not only heat, shock waves, and reactive chemical species but also a jet flow with sub millimeter-sized bubbles. Cells exposed to the plasma exhibited detachment, morphological changes, and changes in the actin cytoskeletal structure. The experimental results suggest that wall shear stress over 160 Pa was generated on the surface of the cells by the plasma. It is one of the main factors that cause those cellular responses. We believe that our findings would provide valuable insight into advancements in medical applications of plasma in a liquid.

  2. Adapting the Medium: Dynamics of Intermedial Adaptation in Contemporary Japanese Popular Visual Culture

    Directory of Open Access Journals (Sweden)

    Pusztai Beáta

    2015-08-01

    Full Text Available With respect to adaptation studies, contemporary Japanese popular culture signifies a unique case, as different types of media (be those textual, auditive, visual or audio-visual are tightly intertwined through the “recycling” of successful characters and stories. As a result, a neatly woven net of intermedial adaptations has been formed - the core of this complex system being the manga-anime-live-action film “adaptational triangle.” On the one hand, the paper addresses the interplay of the various factors by which the very existence of this network is made possible, such as the distinctive cultural attitude to “originality,” the structure of the comics, animation and film industries, and finally, the role of fictitious genealogies of both traditional and contemporary media in the negotiation of national identity. On the other hand, the essay also considers some of the most significant thematic, narrative, and stylistic effects this close interconnectedness has on the individual medium. Special attention is being paid to the nascent trend of merging the adaptive medium with that of the original story (viewing adaptation as integration, apparent in contemporary manga-based live- action comedies, as the extreme case of intermedial adaptation. That is, when the aim of the adaptational process is no longer the transposition of the story but the adaptation (i.e. the incorporation of the medium itself- elevating certain medium-specific devices into transmedial phenomena.

  3. The Effect of Culture Medium on Metabolic and Antibacterial Activities of Probiotic Bacteria

    Directory of Open Access Journals (Sweden)

    Mirdavoudi F

    2012-01-01

    Full Text Available Background and Objectives: Probiotic bacteria is added directly to food components and it has beneficial effect on function and the health of organisms. The bifidogenic factors enter the colon where they contribute to an increase lactic acid bacteria population including Lactobacilli and Bifidobacteria and they inhibit enteric pathogenic bacterial growth. The aim of this study is to investigate the effect of culture medium on metabolic and antibacterial of probiotic bacteria.Methods: In this study, the probiotics bacterial and intestine pathogenic are to be used. Lactobacilli and Bifidobacterium were identified by plating samples on MRS medium, Gram Staining and standard biochemical methods. The effect of antagonistic probiotics was investigated in the presence of growth factor in the method well diffusion Ager on the Shigella flexneri (PTCC 1234, Escherichia coli (PTCC 1552, Salmonella typhi ( PTCC 1609 and the culture medium pH was measured.Results: The probiotics bacterial growth in MRS and lactose1%, sorbitol, raffinose, riboflavin were shown the effect antibacterial. The results of the study show the most antagonistic activity in commercial strain Lactobacillus acidophilus on Shigella flexneri and lower activity was in Lactobacillus casei (PTCC 1608, and Salmonella typhimurium (PTCC 1609, and also in Bbifidobacterium bifidum, it showed the most decrease pH value.Conclusion: According to the result of the study, adding growth factors to MRS medium base and lactose 1%, probiotic growth was increased and which also increased antagonistic activity.

  4. Online games as a medium of cultural communication: An ethnographic study of socio-technical transformation

    OpenAIRE

    Chee, Florence

    2012-01-01

    This dissertation explores the place and meaning of online games in everyday life. In South Korea, online games are a prominent part of popular culture and this medium has come under public criticism for various societal ills, such as Internet addiction and a hopeless dependence upon online games. Humanistic accounts of Information-Communication Technology (ICT) usage are still a minority body of research. All too often, studies of engagement with technology reduce questions to their basic va...

  5. Does supplementation of in-vitro culture medium with melatonin improve IVF outcome in PCOS?

    Science.gov (United States)

    Kim, Mi Kyoung; Park, Eun A; Kim, Hyung Joon; Choi, Won Yun; Cho, Jung Hyun; Lee, Woo Sik; Cha, Kwang Yul; Kim, You Shin; Lee, Dong Ryul; Yoon, Tae Ki

    2013-01-01

    Human pre-ovulatory follicular fluid (FF) contains a higher concentration of melatonin than serum. The aim of this study was to evaluate the effect of melatonin supplementation of culture medium on the clinical outcomes of an in-vitro maturation (IVM) IVF-embryo transfer programme for patients with polycystic ovarian syndrome (PCOS). Melatonin concentrations in the culture media of granulosa cells (GC) or cumulus-oocyte-complexes (COC) were measured and the clinical outcomes after using IVM media with or without melatonin were analysed. In the culture media of GC or COC, melatonin concentrations gradually increased. When human chorionic gonadotrophin priming protocols were used, implantation rates in the melatonin-supplemented group were higher than those of the non-supplemented control group (PPregnancy rates were also higher, although not significantly. The findings suggest that the addition of melatonin to IVM media may improve the cytoplasmic maturation of human immature oocytes and subsequent clinical outcomes. It is speculated that follicular melatonin may be released from luteinizing GC during late folliculogenesis and that melatonin supplementation may be used to improve the clinical outcomes of IVM IVF-embryo transfer. Melatonin is primarily produced by the pineal gland and regulates a variety of important central and peripheral actions related to circadian rhythms and reproduction. Interestingly, human pre-ovulatory follicular fluid contains a higher concentration of melatonin than serum. However, in contrast to animal studies, the direct role of melatonin on oocyte maturation in the human system has not yet been investigated. So, the aim of the study was to evaluate the effect of melatonin supplementation of culture medium on the clinical outcome of an in-vitro maturation (IVM) IVF-embryo transfer programme for PCOS patients. The melatonin concentrations in culture medium of granulosa cells (GC) or cumulus-oocyte-complexes (COC) were measured and the

  6. Effect of explant density and medium culture volumes on cassava micropropagation in Temporal Immersion System

    Directory of Open Access Journals (Sweden)

    Milagros Basail

    2003-04-01

    Full Text Available Due to the need of producing high quality planting material available to cassava growers, it has been necessary to look for alternatives in order to increase the efficiancy of in vitro propagation methods and their automation, such as the use of the Temporal Immersion Systems (RITA®. This work was carried out to increase the multiplication coefficient for cassava mass propagation through out Temporal Immersion Systems. The clone ‘CMC-40’ was used. Different medium volumes per explant, and material density per unit at a given Immersion frequency were tested. The highest results were obtained in the 2.8 multiplication coefficient with 20 ml culture medium volume and 3.2 using a density of 40 explants/flask. When the Temporal Immersion System is used with these results, a more efficient method for cassava micropropagation is established and also higher quality vitroplants for the rooting stage and further acclimatization in field conditions are produced. Key Words: Tissue Culture, liquid culture medium, Manihot esculenta Crantz

  7. Influence of embryo culture medium on incidence of ectopic pregnancy in in vitro fertilization.

    Science.gov (United States)

    Lin, Shengli; Li, Rong; Zheng, Xiaoying; Chi, Hongbin; Ren, Xiulian; Yang, Rui; Liu, Ping; Qiao, Jie

    2015-12-01

    To explore the effect of type of media used to culture embryos for IVF on the incidence of ectopic pregnancy (EP). Retrospective analysis. University-affiliated IVF center. The retrospective analysis involved 23,481 women who underwent IVF-ET cycles between 2011 and 2013. None. There was an association between EP and the culture medium. During 23,481 fresh transfer cycles, 364 patients were diagnosed with EP. The EP to clinical pregnancy rate was 3.01% in the G5 group, 3.89% in the G5 Plus group, and 4.04% in the Global group. The EP to clinical pregnancy rates were significantly higher in the G5 Plus and Global groups than in the G5 group. After adjusting for confounding factors, the incidence of EP was significantly associated with the G5 Plus and Global media. Our results showed that there is an association between incidence of EP and the culture medium. The rates of EP to clinical pregnancy were significantly higher in the G5 Plus and Global media than in the G5 medium. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Effect of cultural conditions on antrodin C production by basidiomycete Antrodia camphorata in solid-state fermentation.

    Science.gov (United States)

    Xia, Yongjun; Wang, Yuanlong; Zhang, Bobo; Xu, Ganrong; Ai, Lianzhong

    2014-01-01

    Antrodia camphorata is a medicinal fungus and antrodin C is one of the main bioactive components of A. camphorata in the submerged fermentation (SmF). To optimize the culture conditions, the factors influencing the production of antrodin C by A. camphorata under solid-state fermentation (SSF) were investigated in this study. Different solid substrates and external nitrogen sources were tested for their efficiency in producing antrodin C. The response surface methodology was applied to evaluate the influence of several variables, namely, the concentrations of soybean meal, initial moisture content, and inoculum density on antrodin C production in solid-state fermentation. The experimental results show that the optimum fermentation medium for antrodin C production by A. camphorata was composed of 0.578 g soybean meal, 0.05 g Na2 HPO4 , 0.05 g MgSO4 for 100 g rice, with 51.83% initial moisture content, 22 day culture time, 28 °C culture temperature, and 35.54% inoculum density. At optimized conditions, 6,617.36 ± 92.71 mg kg(-1) yield of antrodin C was achieved. Solid-state fermentation is one good cultural method to improve the production of antrodin C by A. camphorata. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  9. Fate and effects of octylphenol in a Microcystis aeruginosa culture medium

    Energy Technology Data Exchange (ETDEWEB)

    Baptista, Mafalda S. [CIMAR/CIIMAR, Centro Interdisciplinar de Investigacao Marinha e Ambiental and FCUP, Faculdade de Ciencias, Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto (Portugal)], E-mail: abaptista@fc.up.pt; Stoichev, Teodor; Basto, M. Clara P.; Vasconcelos, Vitor M.; Vasconcelos, M.Teresa S.D. [CIMAR/CIIMAR, Centro Interdisciplinar de Investigacao Marinha e Ambiental and FCUP, Faculdade de Ciencias, Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto (Portugal)

    2009-04-09

    Octylphenol (OP) is a xenobiotic with endocrine disrupting properties found in freshwaters worldwide. Its effects have been studied in organisms with nuclear receptors but effects on phytoplankton communities are poorly characterized, despite the fact that these organisms are constantly exposed to this compound. For this reason fate and effects of OP in the cyanobacterium Microcystis aeruginosa were assessed from 10 nM to 5 {mu}M OP concentration. Up to a test concentration of 250 nM, OP removal increased significantly in the presence of cyanobacteria, the compound half-life in the absence of cells being 15 days against 9 days in the presence of the cells. Only 4% of the total OP removed was found bound to the cells, indicating an active metabolization of the compound. Moreover, the role of the exudates produced by M. aeruginosa, in the OP removal from culture medium, was assessed. Culture medium with exudates, resulting from a 7-day growth of M. aeruginosa, spiked with 50 nM OP, showed a higher half-life (22 days). Compared to culture medium without exudates, it can be hypothesized that higher organic matter concentrations make the hydrolysis or photolysis of OP more difficult. In culture media, the cells of M. aeruginosa could compensate and even counteract this, as OP half-life was shortened. At higher OP levels (1.25 and 5 {mu}M) M. aeruginosa growth was impaired, indicating toxic effects. This shortage of biomass prevented the M. aeruginosa-assisted OP withdrawal from the culture media.

  10. Fate and effects of octylphenol in a Microcystis aeruginosa culture medium

    International Nuclear Information System (INIS)

    Baptista, Mafalda S.; Stoichev, Teodor; Basto, M. Clara P.; Vasconcelos, Vitor M.; Vasconcelos, M.Teresa S.D.

    2009-01-01

    Octylphenol (OP) is a xenobiotic with endocrine disrupting properties found in freshwaters worldwide. Its effects have been studied in organisms with nuclear receptors but effects on phytoplankton communities are poorly characterized, despite the fact that these organisms are constantly exposed to this compound. For this reason fate and effects of OP in the cyanobacterium Microcystis aeruginosa were assessed from 10 nM to 5 μM OP concentration. Up to a test concentration of 250 nM, OP removal increased significantly in the presence of cyanobacteria, the compound half-life in the absence of cells being 15 days against 9 days in the presence of the cells. Only 4% of the total OP removed was found bound to the cells, indicating an active metabolization of the compound. Moreover, the role of the exudates produced by M. aeruginosa, in the OP removal from culture medium, was assessed. Culture medium with exudates, resulting from a 7-day growth of M. aeruginosa, spiked with 50 nM OP, showed a higher half-life (22 days). Compared to culture medium without exudates, it can be hypothesized that higher organic matter concentrations make the hydrolysis or photolysis of OP more difficult. In culture media, the cells of M. aeruginosa could compensate and even counteract this, as OP half-life was shortened. At higher OP levels (1.25 and 5 μM) M. aeruginosa growth was impaired, indicating toxic effects. This shortage of biomass prevented the M. aeruginosa-assisted OP withdrawal from the culture media

  11. Somatic embryogenesis on Musa AAAB, cv. FHIA-18, using liquids culture mediums

    Directory of Open Access Journals (Sweden)

    Luis A. Barranco

    2002-04-01

    Full Text Available Homogenous cell suspensions were iniciated from somatic embryos in the globular stage and the greatest volume of cell biomass on multiplying the suspensions at a density of 3.0% PCV. From the fifteenth day in culture medium for the formation of embryos, structures consisting of proembryos and somatic embryos in the globular stage started to form. With respect to the densities studied, the best results were obtained with 100 mgFW, where 1 871 SE.l-1 formed with a weight of 248 mgFW.l-1 after 30 days. With an initial density of 0.6 gFW in the culture medium for secondary multiplication, an increase of 42.9-fold the initial amount of fresh weight was obtained; after 60 days of culture 15 985 SE.l-1 were obtained. The greatest percentage of maturation was obtained with 400 mgFW with 70% of mature somatic embryos. The positive effect of Biobras-6 (brassinosteroid analogous was confirmed, with a concentration of 0.01 mg.l-1 the best germination percentages were obtained in liquid and semisolid culture medium. Embryo germination in temporaly inmersion (RITA was achieved with an inoculum density of 0.5 gFW for system with 85% germination. One thousand plants obtained from somatic embryos were taken to ex vitro environment, along with plants derived from conventional micropropagation (shoot tips to carry out studies on the possible presence of somaclonal variation. During the first cycle of production, the plants derived from the two methods in vitro culture showed differences with respect to the plants derived from corms in height, diameter and number of suckers. In the second production cycle, the plants from somatic embryos showed similar characteristics to the plants derived from shoot tip and corms with respect to the morphological parameters evaluated, with only 0.2% of the plants with phenotypic changes. Key Words: Banana, cellular density, germination, somaclonal variability, somatic embryo

  12. Improved Murine Blastocyst Quality and Development in a Single Culture Medium Compared to Sequential Culture Media.

    Science.gov (United States)

    Hennings, Justin M; Zimmer, Randall L; Nabli, Henda; Davis, J Wade; Sutovsky, Peter; Sutovsky, Miriam; Sharpe-Timms, Kathy L

    2016-03-01

    Validate single versus sequential culture media for murine embryo development. Prospective laboratory experiment. Assisted Reproduction Laboratory. Murine embryos. Thawed murine zygotes cultured for 3 or 5 days (d3 or d5) in single or sequential embryo culture media developed for human in vitro fertilization. On d3, zygotes developing to the 8 cell (8C) stage or greater were quantified using 4',6-diamidino-2-phenylindole (DAPI), and quality was assessed by morphological analysis. On d5, the number of embryos reaching the blastocyst stage was counted. DAPI was used to quantify total nuclei and inner cell mass nuclei. Localization of ubiquitin C-terminal hydrolase L1 (UCHL1) and ubiquitin C-terminal hydrolase L3 (UCHL3) was reference points for evaluating cell quality. Comparing outcomes in single versus to sequential media, the odds of embryos developing to the 8C stage on d3 were 2.34 time greater (P = .06). On d5, more embryos reached the blastocyst stage (P = culture. Human embryo studies are needed. © The Author(s) 2015.

  13. Development and Validation of a Liquid Medium (M7H9C) for Routine Culture of Mycobacterium avium subsp. paratuberculosis To Replace Modified Bactec 12B Medium

    Science.gov (United States)

    Whittington, Ann-Michele; Waldron, Anna; Begg, Douglas J.; de Silva, Kumi; Purdie, Auriol C.; Plain, Karren M.

    2013-01-01

    Liquid culture of Mycobacterium avium subsp. paratuberculosis from clinical samples, such as feces, is the most sensitive antemortem test for the diagnosis of Johne's disease in ruminants. In Australia, New Zealand, the United States, and some other countries, the Bactec 460 system with modified Bactec 12B medium (Becton, Dickinson) has been the most commonly used liquid culture system, but it was discontinued in 2012. In this study, a new liquid culture medium, M7H9C, was developed. It consists of a Middlebrook 7H9 medium base with added Casitone, albumin, dextrose, catalase, egg yolk, mycobactin J, and a cocktail of antibiotics. We found that polyoxyethylene stearate (POES) was not essential for the cultivation of M. avium subsp. paratuberculosis in either the Bactec 12B or the M7H9C medium. The limit of detection determined using pure cultures of the C and S strains of M. avium subsp. paratuberculosis was 7 bacilli per 50 μl inoculum in the two media. The new medium was validated using 784 fecal and tissue samples from sheep and cattle, >25% of which contained viable M. avium subsp. paratuberculosis. Discrepant results for the clinical samples between the two media were mostly associated with samples that contained <10 viable bacilli per gram, but these results were relatively uncommon, and the performances of the two media were not significantly different. M7H9C medium was less than half the cost of the Bactec 12B medium and did not require regular examination during incubation, but a confirmatory IS900 PCR test had to be performed on every culture after the predetermined incubation period. PMID:24048541

  14. Generation of reactive oxygen species from porous silicon microparticles in cell culture medium.

    Science.gov (United States)

    Low, Suet Peng; Williams, Keryn A; Canham, Leigh T; Voelcker, Nicolas H

    2010-06-01

    Nanostructured (porous) silicon is a promising biodegradable biomaterial, which is being intensively researched as a tissue engineering scaffold and drug-delivery vehicle. Here, we tested the biocompatibility of non-treated and thermally-oxidized porous silicon particles using an indirect cell viability assay. Initial direct cell culture on porous silicon determined that human lens epithelial cells only poorly adhered to non-treated porous silicon. Using an indirect cell culture assay, we found that non-treated microparticles caused complete cell death, indicating that these particles generated a toxic product in cell culture medium. In contrast, thermally-oxidized microparticles did not reduce cell viability significantly. We found evidence for the generation of reactive oxygen species (ROS) by means of the fluorescent probe 2',7'-dichlorofluorescin. Our results suggest that non-treated porous silicon microparticles produced ROS, which interacted with the components of the cell culture medium, leading to the formation of cytotoxic species. Oxidation of porous silicon microparticles not only mitigated, but also abolished the toxic effects.

  15. Evaluation of a Chromogenic Culture Medium for Isolation of Clostridium difficile within 24 Hours ▿

    Science.gov (United States)

    Perry, John D.; Asir, Kerry; Halimi, Diane; Orenga, Sylvain; Dale, Joanne; Payne, Michelle; Carlton, Ruth; Evans, Jim; Gould, F. Kate

    2010-01-01

    Rapid and effective methods for the isolation of Clostridium difficile from stool samples are desirable to obtain isolates for typing or to facilitate accurate diagnosis of C. difficile-associated diarrhea. We report on the evaluation of a prototype chromogenic medium (ID C. difficile prototype [IDCd]) for isolation of C. difficile. The chromogenic medium was compared using (i) 368 untreated stool samples that were also inoculated onto CLO medium, (ii) 339 stool samples that were subjected to alcohol shock and also inoculated onto five distinct selective agars, and (iii) standardized suspensions of 10 C. difficile ribotypes (untreated and alcohol treated) that were also inoculated onto five distinct selective agars. Two hundred thirty-six isolates of C. difficile were recovered from 368 untreated stool samples, and all but 1 of these strains (99.6%) were recovered on IDCd within 24 h, whereas 74.6% of isolates were recovered on CLO medium after 48 h. Of 339 alcohol-treated stool samples cultured onto IDCd and five other selective agars, C. difficile was recovered from 218 samples using a combination of all media. The use of IDCd allowed recovery of 96.3% of isolates within 24 h, whereas 51 to 83% of isolates were recovered within 24 h using the five other media. Finally, when they were challenged with pure cultures, all 10 ribotypes of C. difficile generated higher colony counts on IDCd irrespective of alcohol pretreatment or duration of incubation. We conclude that IDCd is an effective medium for isolation of C. difficile from stool samples within 24 h. PMID:20739493

  16. Formation of industrial mixed culture biofilm in chlorophenol cultivated medium of microbial fuel cell

    Science.gov (United States)

    Hassan, Huzairy; Jin, Bo; Dai, Sheng; Ngau, Cornelius

    2016-11-01

    The formation of microbial biofilm while maintaining the electricity output is a challenging topic in microbial fuel cell (MFC) studies. This MFC critical factor becomes more significant when handling with industrial wastewater which normally contains refractory and toxic compounds. This study explores the formation of industrial mixed culture biofilm in chlorophenol cultivated medium through observing and characterizing microscopically its establishment on MFC anode surface. The mixed culture was found to develop its biofilm on the anode surface in the chlorophenol environment and established its maturity and dispersal stages with concurrent electricity generation and phenolic degradation. The mixed culture biofilm engaged the electron transfer roles in MFC by generating current density of 1.4 mA/m2 and removing 53 % of 2,4-dichlorophenol. The results support further research especially on hazardous wastewater treatment using a benign and sustainable method.

  17. Profiling of Extracellular Toxins Associated with Diarrhetic Shellfish Poison in Prorocentrum lima Culture Medium by High-Performance Liquid Chromatography Coupled with Mass Spectrometry

    Science.gov (United States)

    Pan, Lei; Chen, Junhui; Shen, Huihui; He, Xiuping; Li, Guangjiu; Song, Xincheng; Zhou, Deshan; Sun, Chengjun

    2017-01-01

    Extracellular toxins released by marine toxigenic algae into the marine environment have attracted increasing attention in recent years. In this study, profiling, characterization and quantification of extracellular toxin compounds associated with diarrhetic shellfish poison (DSP) in the culture medium of toxin-producing dinoflagellates were performed using high-performance liquid chromatography–high-resolution mass spectrometry/tandem mass spectrometry for the first time. Results showed that solid-phase extraction can effectively enrich and clean the DSP compounds in the culture medium of Prorocentrum lima (P. lima), and the proposed method achieved satisfactory recoveries (94.80%–100.58%) and repeatability (relative standard deviation ≤9.27%). Commercial software associated with the accurate mass information of known DSP toxins and their derivatives was used to screen and identify DSP compounds. Nine extracellular DSP compounds were identified, of which seven toxins (including OA-D7b, OA-D9b, OA-D10a/b, and so on) were found in the culture medium of P. lima for the first time. The results of quantitative analysis showed that the contents of extracellular DSP compounds in P. lima culture medium were relatively high, and the types and contents of intracellular and extracellular toxins apparently varied in the different growth stages of P. lima. The concentrations of extracellular okadaic acid and dinophysistoxin-1 were within 19.9–34.0 and 15.2–27.9 μg/L, respectively. The total concentration of the DSP compounds was within the range of 57.70–79.63 μg/L. The results showed that the proposed method is an effective tool for profiling the extracellular DSP compounds in the culture medium of marine toxigenic algae. PMID:28974018

  18. Profiling of Extracellular Toxins Associated with Diarrhetic Shellfish Poison in Prorocentrum lima Culture Medium by High-Performance Liquid Chromatography Coupled with Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Lei Pan

    2017-09-01

    Full Text Available Extracellular toxins released by marine toxigenic algae into the marine environment have attracted increasing attention in recent years. In this study, profiling, characterization and quantification of extracellular toxin compounds associated with diarrhetic shellfish poison (DSP in the culture medium of toxin-producing dinoflagellates were performed using high-performance liquid chromatography–high-resolution mass spectrometry/tandem mass spectrometry for the first time. Results showed that solid-phase extraction can effectively enrich and clean the DSP compounds in the culture medium of Prorocentrum lima (P. lima, and the proposed method achieved satisfactory recoveries (94.80%–100.58% and repeatability (relative standard deviation ≤9.27%. Commercial software associated with the accurate mass information of known DSP toxins and their derivatives was used to screen and identify DSP compounds. Nine extracellular DSP compounds were identified, of which seven toxins (including OA-D7b, OA-D9b, OA-D10a/b, and so on were found in the culture medium of P. lima for the first time. The results of quantitative analysis showed that the contents of extracellular DSP compounds in P. lima culture medium were relatively high, and the types and contents of intracellular and extracellular toxins apparently varied in the different growth stages of P. lima. The concentrations of extracellular okadaic acid and dinophysistoxin-1 were within 19.9–34.0 and 15.2–27.9 μg/L, respectively. The total concentration of the DSP compounds was within the range of 57.70–79.63 μg/L. The results showed that the proposed method is an effective tool for profiling the extracellular DSP compounds in the culture medium of marine toxigenic algae.

  19. Profiling of Extracellular Toxins Associated with Diarrhetic Shellfish Poison in Prorocentrum lima Culture Medium by High-Performance Liquid Chromatography Coupled with Mass Spectrometry.

    Science.gov (United States)

    Pan, Lei; Chen, Junhui; Shen, Huihui; He, Xiuping; Li, Guangjiu; Song, Xincheng; Zhou, Deshan; Sun, Chengjun

    2017-09-30

    Extracellular toxins released by marine toxigenic algae into the marine environment have attracted increasing attention in recent years. In this study, profiling, characterization and quantification of extracellular toxin compounds associated with diarrhetic shellfish poison (DSP) in the culture medium of toxin-producing dinoflagellates were performed using high-performance liquid chromatography-high-resolution mass spectrometry/tandem mass spectrometry for the first time. Results showed that solid-phase extraction can effectively enrich and clean the DSP compounds in the culture medium of Prorocentrum lima ( P. lima ), and the proposed method achieved satisfactory recoveries (94.80%-100.58%) and repeatability (relative standard deviation ≤9.27%). Commercial software associated with the accurate mass information of known DSP toxins and their derivatives was used to screen and identify DSP compounds. Nine extracellular DSP compounds were identified, of which seven toxins (including OA-D7b, OA-D9b, OA-D10a/b, and so on) were found in the culture medium of P. lima for the first time. The results of quantitative analysis showed that the contents of extracellular DSP compounds in P. lima culture medium were relatively high, and the types and contents of intracellular and extracellular toxins apparently varied in the different growth stages of P. lima . The concentrations of extracellular okadaic acid and dinophysistoxin-1 were within 19.9-34.0 and 15.2-27.9 μg/L, respectively. The total concentration of the DSP compounds was within the range of 57.70-79.63 μg/L. The results showed that the proposed method is an effective tool for profiling the extracellular DSP compounds in the culture medium of marine toxigenic algae.

  20. Influence of embryo culture medium (G5 and HTF) on pregnancy and perinatal outcome after IVF: a multicenter RCT

    NARCIS (Netherlands)

    Kleijkers, S.H.; Mantikou, E.; Slappendel, E.; Consten, D.; Echten-Arends, J. van; Wetzels, A.M.M.; Wely, M. van; Smits, L.J.; Montfoort, A.P. van; Repping, S.; Dumoulin, J.C.; Mastenbroek, S.

    2016-01-01

    STUDY QUESTION: Does embryo culture medium influence pregnancy and perinatal outcome in IVF? SUMMARY ANSWER: Embryo culture media used in IVF affect treatment efficacy and the birthweight of newborns. WHAT IS KNOWN ALREADY: A wide variety of culture media for human preimplantation embryos in

  1. Influence of embryo culture medium (G5 and HTF) on pregnancy and perinatal outcome after IVF: a multicenter RCT

    NARCIS (Netherlands)

    Kleijkers, Sander H. M.; Mantikou, Eleni; Slappendel, Els; Consten, Dimitri; van Echten-Arends, Jannie; Wetzels, Alex M.; van Wely, Madelon; Smits, Luc J. M.; van Montfoort, Aafke P. A.; Repping, Sjoerd; Dumoulin, John C. M.; Mastenbroek, Sebastiaan

    2016-01-01

    Does embryo culture medium influence pregnancy and perinatal outcome in IVF? Embryo culture media used in IVF affect treatment efficacy and the birthweight of newborns. A wide variety of culture media for human preimplantation embryos in IVF/ICSI treatments currently exists. It is unknown which

  2. Influence of embryo culture medium (G5 and HTF) on pregnancy and perinatal outcome after IVF : a multicenter RCT

    NARCIS (Netherlands)

    Kleijkers, Sander H. M.; Mantikou, Eleni; Slappendel, Els; Consten, Dimitri; van Echten - Arends, Jannie; Wetzels, Alex M.; van Wely, Madelon; Smits, Luc J. M.; van Montfoort, Aafke P. A.; Repping, Sjoerd; Dumoulin, John C. M.; Mastenbroek, Sebastiaan

    2016-01-01

    Does embryo culture medium influence pregnancy and perinatal outcome in IVF? Embryo culture media used in IVF affect treatment efficacy and the birthweight of newborns. A wide variety of culture media for human preimplantation embryos in IVF/ICSI treatments currently exists. It is unknown which

  3. Organizational Culture and Open Innovation Performance in Small and Medium-sized Enterprises (SMEs in Poland

    Directory of Open Access Journals (Sweden)

    Mazur Jolanta

    2016-09-01

    Full Text Available This study investigates the links between organizational culture, the use of open innovation sources and the performance of SMEs. The main hypothesis of the study is that a special type of organizational culture (termed innovative culture, which fosters creativity, learning and inter-employee cooperation – will correspond with a greater scope of open innovation sources and higher levels of innovative, operational and financial performance. The study was based on a representative CATI survey of 473 SMEs operating in manufacturing and services industries in Poland. Our statistical analysis relied on building and testing structural equation model with the AMOS software. The findings confirmed a positive association between innovative culture and the scope of open sources of innovation. However, innovative culture had no direct effect on the percentage of sales from new and modified products, which is often used as a metric of innovativeness, but did show a positive influence on an index of operational performance and ROI. Such statistical patterns suggest that fostering innovative culture is beneficial to a company, though probably not through an increased number of product innovations, but rather via process, administrative and marketing innovations, as well as other gains in efficiency attained due to more streamlined employee cooperation and knowledge exchange. The study adds to the existing body of knowledge in management science by providing a better understanding of mechanisms underlying innovative culture’s impacts on open innovation practices and metrics of operational and financial performance in the context of small and medium enterprises.

  4. Effect of medium components and culture conditions in Bacillus subtilis EA-CB0575 spore production.

    Science.gov (United States)

    Posada-Uribe, Luisa F; Romero-Tabarez, Magally; Villegas-Escobar, Valeska

    2015-10-01

    Bacillus subtilis spores have important biotechnological applications; however, achieving both, high spore cell densities and sporulation efficiencies in fermentation, is poorly reported. In this study, medium components and culture conditions were optimized with different statistical methods to increase spore production of the plant growth promoting rhizobacteria B. subtilis EA-CB0575. Key medium components were determined with Plackett-Burman (PB) design, and the optimum concentration levels of two components (glucose, MgSO4·7H2O) were optimized with a full factorial and central composite design, achieving 1.37 × 10(9) CFU/mL of spore cell density and 93.5 % of sporulation efficiency in shake flask. The optimized medium was used to determine the effect of culture conditions on spore production at bioreactor level, finding that maintaining pH control did not affect significantly spore production, while the interaction of agitation and aeration rates had a significant effect on spore cell density. The overall optimization generated a 17.2-fold increase in spore cell density (8.78 × 10(9) CFU/mL) and 1.9-fold increase in sporulation efficiency (94.2 %) compared to that of PB design. These results indicate the potential of B. subtilis EA-CB0575 to produce both, high spore cell densities and sporulation efficiencies, with very low nutrient requirements and short incubation period which can represent savings of process production.

  5. Using of acoustic technologies for detection of explosives in gas, liquid and solid medium

    International Nuclear Information System (INIS)

    Valyaev, A. N.; Yanushkevich, V.A.

    2004-01-01

    contained acoustic radiator and detector, with probe selection of diagnostic material. That is why at first the acoustic passports will be obtained for all types of explosives and their data bank will be formed. This bank will included C (ω,T), α (ω,T) and ρ (ω,T) dependences, which will identify a presence and a specific type of explosive. We propose the universal system, that may be quickly adapted for search and diagnostics of different types of matter, including a qualitative and quantitative analysis of their few main components. This system may be realized in the development of devices, that used the acoustic techniques. The principles of system function is discussed. The practical realization of this method on example of image of alcohol beverage type. The presented 2a and 3D diagrams for system of ethyl alcohol-sugar-water solutions proved the possibilities of our technique for the operative and reliable diagnostic. The similar technique we successfully used in the development and the application of new devices for monitoring and measurements of alcohol and sugar concentrations in industrial flood plains of vodka, brandies, liqueurs, wines and champagnes at Russian plants. The principle advantages and disadvantages of the proposed technique, including the special engineering and design elements of the devices for diagnostic of explosives are under consideration. It is noticed, that this technique may be used for detection of solid and liquid explosives in grounds and in another solid materials, in water and gas mediums and we recommend to use it as the special monitoring method for very important large objects, such as dams of uranium tailing storages in Tyan-Shan Mountains

  6. A SIMPLE METHOD TO CONTROL THE MOISTURE CONTENT OF THE FERMENTING MEDIUM DURING LABORATORY-SCALE SOLID-STATE FERMENTATION EXPERIMENTS

    Directory of Open Access Journals (Sweden)

    W. BORZANI

    1999-03-01

    Full Text Available When the moisture content of the fermenting medium significantly decreases during laboratory-scale solid-state fermentation tests, the quantity of water to be periodically added to the medium in order to control its moisture content may be evaluated from the water evaporation rate of the non-inoculated medium.

  7. Effect of cell culture medium components on color of formulated monoclonal antibody drug substance.

    Science.gov (United States)

    Vijayasankaran, Natarajan; Varma, Sharat; Yang, Yi; Mun, Melissa; Arevalo, Silvana; Gawlitzek, Martin; Swartz, Trevor; Lim, Amy; Li, Feng; Zhang, Boyan; Meier, Steve; Kiss, Robert

    2013-01-01

    As the industry moves toward subcutaneous delivery as a preferred route of drug administration, high drug substance concentrations are becoming the norm for monoclonal antibodies. At such high concentrations, the drug substance may display a more intense color than at the historically lower concentrations. The effect of process conditions and/or changes on color is more readily observed in the higher color, high concentration formulations. Since color is a product quality attribute that needs to be controlled, it is useful to study the impact of process conditions and/or modifications on color. This manuscript summarizes cell culture experiments and reports on findings regarding the effect of various media components that contribute to drug substance color for a specific monoclonal antibody. In this work, lower drug substance color was achieved via optimization of the cell culture medium. Specifically, lowering the concentrations of B-vitamins in the cell culture medium has the effect of reducing color intensity by as much as 25%. In addition, decreasing concentration of iron was also directly correlated color intensity decrease of as much as 37%. It was also shown that the color of the drug substance directly correlates with increased acidic variants, especially when increased iron levels cause increased color. Potential mechanisms that could lead to antibody coloration are briefly discussed. © 2013 American Institute of Chemical Engineers.

  8. Medium term municipal solid waste generation prediction by autoregressive integrated moving average

    International Nuclear Information System (INIS)

    Younes, Mohammad K.; Nopiah, Z. M.; Basri, Noor Ezlin A.; Basri, Hassan

    2014-01-01

    Generally, solid waste handling and management are performed by municipality or local authority. In most of developing countries, local authorities suffer from serious solid waste management (SWM) problems and insufficient data and strategic planning. Thus it is important to develop robust solid waste generation forecasting model. It helps to proper manage the generated solid waste and to develop future plan based on relatively accurate figures. In Malaysia, solid waste generation rate increases rapidly due to the population growth and new consumption trends that characterize the modern life style. This paper aims to develop monthly solid waste forecasting model using Autoregressive Integrated Moving Average (ARIMA), such model is applicable even though there is lack of data and will help the municipality properly establish the annual service plan. The results show that ARIMA (6,1,0) model predicts monthly municipal solid waste generation with root mean square error equals to 0.0952 and the model forecast residuals are within accepted 95% confident interval

  9. Medium term municipal solid waste generation prediction by autoregressive integrated moving average

    Science.gov (United States)

    Younes, Mohammad K.; Nopiah, Z. M.; Basri, Noor Ezlin A.; Basri, Hassan

    2014-09-01

    Generally, solid waste handling and management are performed by municipality or local authority. In most of developing countries, local authorities suffer from serious solid waste management (SWM) problems and insufficient data and strategic planning. Thus it is important to develop robust solid waste generation forecasting model. It helps to proper manage the generated solid waste and to develop future plan based on relatively accurate figures. In Malaysia, solid waste generation rate increases rapidly due to the population growth and new consumption trends that characterize the modern life style. This paper aims to develop monthly solid waste forecasting model using Autoregressive Integrated Moving Average (ARIMA), such model is applicable even though there is lack of data and will help the municipality properly establish the annual service plan. The results show that ARIMA (6,1,0) model predicts monthly municipal solid waste generation with root mean square error equals to 0.0952 and the model forecast residuals are within accepted 95% confident interval.

  10. Medium term municipal solid waste generation prediction by autoregressive integrated moving average

    Energy Technology Data Exchange (ETDEWEB)

    Younes, Mohammad K.; Nopiah, Z. M.; Basri, Noor Ezlin A.; Basri, Hassan [Department of Civil and Structural Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia)

    2014-09-12

    Generally, solid waste handling and management are performed by municipality or local authority. In most of developing countries, local authorities suffer from serious solid waste management (SWM) problems and insufficient data and strategic planning. Thus it is important to develop robust solid waste generation forecasting model. It helps to proper manage the generated solid waste and to develop future plan based on relatively accurate figures. In Malaysia, solid waste generation rate increases rapidly due to the population growth and new consumption trends that characterize the modern life style. This paper aims to develop monthly solid waste forecasting model using Autoregressive Integrated Moving Average (ARIMA), such model is applicable even though there is lack of data and will help the municipality properly establish the annual service plan. The results show that ARIMA (6,1,0) model predicts monthly municipal solid waste generation with root mean square error equals to 0.0952 and the model forecast residuals are within accepted 95% confident interval.

  11. Human dental pulp stem cells cultured in serum-free supplemented medium

    Directory of Open Access Journals (Sweden)

    Virginie eBonnamain

    2013-12-01

    Full Text Available Growing evidence show that human dental pulp stem cells (DPSCs could provide a source of adult stem cells for the treatment of neurodegenerative pathologies. In this study, DPSCs were expanded and cultured with a protocol generally used for the culture of neural stem/progenitor cells.Methodology: DPSC cultures were established from third molars. The pulp tissue was enzymatically digested and cultured in serum-supplemented basal medium for 12 hours. Adherent (ADH and non-adherent (non-ADH cell populations were separated according to their differential adhesion to plastic and then cultured in serum-free defined N2 medium with epidermal growth factor (EGF and basic fibroblast growth factor (bFGF. Both ADH and non-ADH populations were analyzed by FACS and/or PCR.Results: FACS analysis of ADH-DPSCs revealed the expression of the mesenchymal cell marker CD90, the neuronal marker CD56, the transferrin receptor CD71, and the chemokine receptor CXCR3, whereas hematopoietic stem cells markers CD45, CD133 and CD34 were not expressed. ADH-DPSCs expressed transcripts coding for the Nestin gene, whereas expression levels of genes coding for the neuronal markers β-III tubulin and NF-M, and the oligodendrocyte marker PLP-1 were donor dependent. ADH-DPSCs did not express the transcripts for GFAP, an astrocyte marker. Cells of the non-ADH population that grew as spheroids expressed Nestin, β-III tubulin, NF-M and PLP-1 transcripts. DPSCs migrated out of the spheroids exhibited an odontoblast-like morphology and expressed a higher level of DSPP and osteocalcin transcripts than ADH-DPSCs. Conclusion: Collectively, these data indicate that human DPSCs can be expended and cultured in serum-free supplemented medium with EGF and bFGF. ADH-DPSCs and non-ADH populations contained neuronal and/or oligodendrocyte precursors at different stages of commitment and interestingly, cells from spheroid structures seem to be more engaged into the odontoblastic lineage than the

  12. Resource efficiency and culture--workplace training for small and medium-sized enterprises.

    Science.gov (United States)

    Bliesner, Anna; Liedtke, Christa; Rohn, Holger

    2014-05-15

    Although there are already some qualification offers available for enterprises to support resource efficiency innovations, the high potentials that can be identified especially for small and medium sized enterprises (SMEs) have not been activated until now. As successful change lies in the hands of humans, the main aim of vocational education has to be the promotion of organisational and cultural changes in the enterprises. As there is already a small but increasing number of enterprises that perform very well in resource efficiency innovations one question arises: What are typical characteristics of those enterprises? Leaning on a good-practice approach, the project "ResourceCulture" is going to prove or falsify the hypothesis that enterprises being successful with resource efficiency innovations have a specific culture of trust, which substantially contributes to innovation processes, or even initially enables them. Detailed empirical field research will light up which correlations between resource efficiency, innovation and cultures of trust can be found and will offer important aspects for the improvement of management instruments and qualification concepts for workplace training. The project seizes qualification needs that were likewise mentioned by enterprises and consultants, regarding the implementation of resource efficiency. This article - based on first empirical field research results - derives preliminary indications for the design of the qualification module for the target groups resource efficiency consultants and managers. On this basis and in order to implement "ResourceCulture" conceptual and methodological starting points for workplace training are outlined. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. HepG2 cells develop signs of riboflavin deficiency within four days of culture in riboflavin-deficient medium*

    OpenAIRE

    Werner, Ricarda; Manthey, Karoline C.; Griffin, Jacob B.; Zempleni, Janos

    2005-01-01

    Flavin mononucleotide and flavin adenine dinucleotide are essential coenzymes in redox reactions. For example, flavin adenine dinucleotide is a coenzyme for both glutathione reductase and enzymes that mediate the oxidative folding of secretory proteins. Here we investigated short-term effects of moderately riboflavin-deficient culture medium on flavin-related responses in HepG2 hepatocarcinoma cells. Cells were cultured in riboflavin-deficient (3.1 nmol/L) medium for up to six days; controls ...

  14. A novel culture medium designed for the simultaneous enhancement of biomass and lipid production by Chlorella vulgaris UTEX 26.

    Science.gov (United States)

    Ramírez-López, Citlally; Chairez, Isaac; Fernández-Linares, Luis

    2016-07-01

    A novel culture medium to enhance the biomass and lipid production simultaneously by Chlorella vulgaris UTEX 26 was designed in three stages of optimization. Initially, a culture medium was inferred applying the response surface method to adjust six factors [NaNO3, NH4HCO3, MgSO4·7H2O, KH2PO4, K2HPO4 and (NH4)2HPO4], which were selected on the basement of BBM (Bold's Basal Medium) and HAMGM (Highly Assimilable Minimal Growth Medium) culture media. Afterwards, the nitrogen source compound was optimized to reduce both, ammonium and nitrate concentrations. As result of the optimization process, the proposed culture medium improved 40% the biomass (0.73gL(-1)) compared with the BBM medium and 85% the lipid concentration (281mgL(-1)), with respect to HAMGM medium. Some culture media components concentrations were reduced up to 50%. Gas chromatography analysis revealed that C16:0, C18:0, C18:1, C18:2 and C18:3 were the major fatty acids produced by C. vulgaris UTEX 26. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. In vitro propagation of Morus alba L. in semisolid culture medium.

    Directory of Open Access Journals (Sweden)

    Enrique Salas Barbosa

    2005-04-01

    Full Text Available Apical buds as explants were used with the objective to propagate in vitro mulberry plants in semisolid MS culture medium suplemented with 6-BAP and KIN in their establishment and, with different combinations of 6-BAP with ANA in the multiplication phase. In vitro plants were evaluated during the acclimatization phase. It is necessary to supplement the basal MS culture media with 0.5 mg.l-1 of 6-BAP to induce the sprouting and, 0.5 mg.l-1 of 6-BAP and 0.5 mg.l-1 of ANA to multiply the mulberry by nodal segments. In the acclimatization phase a 95% of survival, 30.2 cm of height, 9.8 leaves and 2.02 g.plant-1 of dry mass was observed. In vitro propagation of mulberry was achieved as an alternative for plants production. Key words: acclimatization, apical buds, establishment, explant, shooting

  16. Optimization of culture medium for heavy-ion irradiation bread yeast design

    International Nuclear Information System (INIS)

    Ma Liang; Wang Jufang; Lu Dong; Li Wenjian; Xiao Guoqing

    2013-01-01

    A mutant bread yeast strain with high protein content of 55% was gained by use of "1"2C"6"+ ions. The MINITAB 16.0 software, Plackett-Burman experimental design and response surface methodology were applied to optimize the culture medium for the irradiated yeast. The most important three factors which influenced the culture results were identified as glucose, magnesium sulphate and yeast extract. The path of the steepest ascent was undertaken to approach the optimal region of the three significant factors. Box-Behnken design and response surface methodology were used for the regression analysis. Finally, the optimal fermentation conditions were identified as glucose 11.03 g/L, yeast extract 6.53 g/L and magnesium sulphate 5.59 g/L by the regression analysis. It was found that the biomass of the bread yeasts reached 4.84 g/L and increased by 15% compared to original conditions. (authors)

  17. Biological effects of low doses of ionizing radiations. Evidence of effect of pre-irradiation of culture medium on subsequent growth in Cyanobacterium Synechococcus lividus in culture

    International Nuclear Information System (INIS)

    Conter, A.; Planel, H.

    1986-01-01

    In order to distinguish the direct effects of low dose of ionizing radiations at the cellular level from those indirect through the culture medium, we have compared proliferation of Synechococcus lividus grown in pre-irradiated medium to proliferation of cultures grown in non-irradiated medium. A stimulation of growth was observed at the 7th day in cultures inoculated with cells selected in deceleration phase, while an inhibition occured in cultures inoculated with exponential growing cells. Addition of catalase (100 U/ml) counteracted the stimulating effect but did not change the inhibiting effect induced by pre-irradiated medium. Results demonstrated the indirect effect of low dose of irradiation, implying hydrogen peroxide, but let us to think that others radioproduced products could be also involved in the mechanism [fr

  18. In Vitro Selection of Peanut Somatic Embryos on Medium Containing Culture Filtrate of Sclerotium rolfsii and Plantlet Regeneration

    Directory of Open Access Journals (Sweden)

    YUSNITA

    2005-06-01

    Full Text Available Attempts to identify somaclonal variants of peanut with resistance to Sclerotium stem rot disease due to infection of S. rolfsii were conducted. The objectives of this study were to develop in vitro selection method using culture filtrates of S. rolfsii, identify culture filtrate-insensitive somatic embryo (SE of peanut after in vitro selection and regenerate peanut R0 lines originated from culture filtrate-insensitive SE. To achieve these objectives, peanut embryogenic tissues were cultured on selective medium containing various concentrations of S. rolfsii culture filtrates and sublethal concentration of the filtrates. Medium containing sublethal level of S. rolfsii culture filtrates was used to identify culture filtrate-insensitive SE of peanut. Subsequently, the selected SEs were germinated, plantlets were regenerated and preliminary tested against S. rolfsii. Results of the experiments showed that addition of S. rolfsii culture filtrates into medium for inducing peanut somatic embryos drastically reduced their growth and proliferation. S. rolfsii culture filtrates at 10% concentration has significantly reduced the number of proliferated SE per explant. However, sublethal level was achieved at 30% of culture filtrates concentration. Responses of five peanut cultivars against 30% of culture filtrates were similar, indicating they were similar in their susceptibility against S. rolfsii. A number of culture filtrate-insensitive SE were identified after culturing 1500 clumps of embryogenic tissue of peanut cv. Kelinci for three consecutive passages on medium containing 30% of culture filtrates. Germination of selected SE and regeneration of plantlet from culture filtrate-insensitive SE resulted in 50 peanut R0 lines. These lines have been grown in the plastic house and produced normal seeds for further evaluation. Results of S. rolfsii inoculation indicated the existence of chimera for insensitivity against S. rolfsii.

  19. Low versus high volume of culture medium during embryo transfer: a randomized clinical trial.

    Science.gov (United States)

    Sigalos, George Α; Michalopoulos, Yannis; Kastoras, Athanasios G; Triantafyllidou, Olga; Vlahos, Nikos F

    2018-04-01

    The aim of this prospective randomized control trial was to evaluate if the use of two different volumes (20-25 vs 40-45 μl) of media used for embryo transfer affects the clinical outcomes in fresh in vitro fertilization (IVF) cycles. In total, 236 patients were randomized in two groups, i.e., "low volume" group (n = 118) transferring the embryos with 20-25 μl of medium and "high volume" group (n = 118) transferring the embryos with 40-45 μl of medium. The clinical pregnancy, implantation, and ongoing pregnancy rates were compared between the two groups. No statistically significant differences were observed in clinical pregnancy (46.8 vs 54.3%, p = 0.27), implantation (23.7 vs 27.8%, p = 0.30), and ongoing pregnancy (33.3 vs 40.0%, p = 0.31) rates between low and high volume group, respectively. Higher volume of culture medium to load the embryo into the catheter during embryo transfer does not influence the clinical outcome in fresh IVF cycles. NCT03350646.

  20. Nanoparticle growth and surface chemistry changes in cell-conditioned culture medium.

    Science.gov (United States)

    Kendall, Michaela; Hodges, Nikolas J; Whitwell, Harry; Tyrrell, Jess; Cangul, Hakan

    2015-02-05

    When biomolecules attach to engineered nanoparticle (ENP) surfaces, they confer the particles with a new biological identity. Physical format may also radically alter, changing ENP stability and agglomeration state within seconds. In order to measure which biomolecules are associated with early ENP growth, we studied ENPs in conditioned medium from A549 cell culture, using dynamic light scattering (DLS) and linear trap quadrupole electron transfer dissociation mass spectrometry. Two types of 100 nm polystyrene particles (one uncoated and one with an amine functionalized surface) were used to measure the influence of surface type. In identically prepared conditioned medium, agglomeration was visible in all samples after 1 h, but was variable, indicating inter-sample variability in secretion rates and extracellular medium conditions. In samples conditioned for 1 h or more, ENP agglomeration rates varied significantly. Agglomerate size measured by DLS was well correlated with surface sequestered peptide number for uncoated but not for amine coated polystyrene ENPs. Amine-coated ENPs grew much faster and into larger agglomerates associated with fewer sequestered peptides, but including significant sequestered lactose dehydrogenase. We conclude that interference with extracellular peptide balance and oxidoreductase activity via sequestration is worthy of further study, as increased oxidative stress via this new mechanism may be important for cell toxicity. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  1. Influence of culture medium composition on relative mRNA abundances in domestic cat embryos.

    Science.gov (United States)

    Hribal, R; Jewgenow, K; Braun, B C; Comizzoli, P

    2013-04-01

    Different culture conditions have been used to produce domestic cat embryos. As part of the in vitro procedures, the medium composition significantly affects the quality of the embryo development also. Quality assessments based on cleavage kinetics and blastomere symmetry are useful, but embryos also can differ in their relative gene expression patterns despite similar morphological characteristics. The aim of this study was to compare cat embryos produced with two different in vitro culture systems routinely used in two different laboratories [Smithsonian Conservation Biology Institute, Washington D.C., USA (SCBI) and Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany (IZW)]. Specifically, relative mRNA expression patterns of critical genes for pre-implantation embryo development were assessed in both conditions. Embryos were produced in parallel in both culture systems by IVF using frozen-thawed ejaculated semen in the United States and fresh epididymal sperm in Germany. Success of embryo development in vitro was recorded as well as relative mRNA abundances [DNA methyltransferases 1 and 3A (DNMT1, DNMT3A), gap junction protein alpha 1 (GJA1), octamer-binding transcription factor 4 [OCT4], insulin-like growth factors 1 and 2 receptors (IGF1R, IGF2R), beta-actin (ACTB)] in pools of days 4-5 morulae by semi-quantitative RT-PCR assay. Percentages of cleaved embryos were similar (p > 0.05) between both culture systems, regardless of the location. OCT4 mRNA abundance was higher (p culture system compared with those from the IZW system when epididymal sperm was used for IVF. No clear correlation between the expression pattern and the culture system could be found for all other genes. It is suggested that OCT4 expression might be affected by the media composition in some conditions and can be the indicator of a better embryo quality. © 2012 Blackwell Verlag GmbH.

  2. A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

    Energy Technology Data Exchange (ETDEWEB)

    Aslanova, Afag [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Takagi, Ryo; Yamato, Masayuki; Okano, Teruo [Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Yamamoto, Masakazu, E-mail: yamamoto.ige@twmu.ac.jp [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan)

    2015-05-01

    With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

  3. A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

    International Nuclear Information System (INIS)

    Aslanova, Afag; Takagi, Ryo; Yamato, Masayuki; Okano, Teruo; Yamamoto, Masakazu

    2015-01-01

    With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

  4. Accumulation of 137Cs in trefoil (leaf and stem), ''Mitsuba'', Cryptotaenia japonica Hassk, immersed in hydroponic culture medium

    International Nuclear Information System (INIS)

    Motegi, Misako; Miyake, Sadaaki; Ohsawa, Takashi; Nakazawa, Kiyoaki; Izumo, Yoshiro

    1998-01-01

    Accumulation of 137 Cs in trefoil (leaf and stem), ''Mitsuba'', Cryptotaenia japonica Hassk, with or without root was investigated to prepare higher radioactive plant in hydroponic culture medium (140-150 Bq/ml). It was found that 137 Cs concentration in plant tissue was increased with time, as high as 1.6 times of that in the culture medium after 4 days. On the other hand, 137 Cs concentration was affected by carrier element (Cs>6 ppm) and coexistent elements in the medium. Radioactivity of the plant after 4 days was shown to be sufficient for successive experiments. (author)

  5. Accumulation of {sup 137}Cs in trefoil (leaf and stem), ``Mitsuba``, Cryptotaenia japonica Hassk, immersed in hydroponic culture medium

    Energy Technology Data Exchange (ETDEWEB)

    Motegi, Misako; Miyake, Sadaaki; Ohsawa, Takashi; Nakazawa, Kiyoaki [Saitama Institute of Public Health, Urawa (Japan); Izumo, Yoshiro

    1998-11-01

    Accumulation of {sup 137}Cs in trefoil (leaf and stem), ``Mitsuba``, Cryptotaenia japonica Hassk, with or without root was investigated to prepare higher radioactive plant in hydroponic culture medium (140-150 Bq/ml). It was found that {sup 137}Cs concentration in plant tissue was increased with time, as high as 1.6 times of that in the culture medium after 4 days. On the other hand, {sup 137}Cs concentration was affected by carrier element (Cs>6 ppm) and coexistent elements in the medium. Radioactivity of the plant after 4 days was shown to be sufficient for successive experiments. (author)

  6. Enhanced solubility and bioavailability of sibutramine base by solid dispersion system with aqueous medium.

    Science.gov (United States)

    Li, Dong Xun; Jang, Ki-Young; Kang, Wonku; Bae, Kyoungjin; Lee, Mann Hyung; Oh, Yu-Kyoung; Jee, Jun-Pil; Park, Young-Joon; Oh, Dong Hoon; Seo, Youn Gee; Kim, Young Ran; Kim, Jong Oh; Woo, Jong Soo; Yong, Chul Soon; Choi, Han-Gon

    2010-01-01

    To develop a novel sibutramine base-loaded solid dispersion with improved solubility bioavailability, various solid dispersions were prepared with water, hydroxypropylmethyl cellulose (HPMC), poloxamer and citric acid using spray-drying technique. The effect of HPMC, poloxamer and citric acid on the aqueous solubility of sibutramine was investigated. The physicochemical properties of solid dispersion were investigated using scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and X-ray powder diffraction. The dissolution and pharmacokinetics in rats of solid dispersion were evaluated compared to the sibutramine hydrochloride monohydrate-loaded commercial product (Reductil). The sibutramine base-loaded solid dispersion gave two type forms. Like conventional solid dispersion system, one type appeared as a spherical shape with smooth surface, as the carriers and drug with relatively low melting point were soluble in water and formed it. The other appeared as an irregular form with relatively rough surface. Unlike conventional solid dispersion system, this type changed no crystalline form of drug. Our results suggested that this type was formed by attaching hydrophilic carriers to the surface of drug without crystal change, resulting from changing the hydrophobic drug to hydrophilic form. The sibutramine-loaded solid dispersion at the weight ratio of sibutramine base/HPMC/poloxamer/citric acid of 5/3/3/0.2 gave the maximum drug solubility of about 3 mg/ml. Furthermore, it showed the similar plasma concentration, area under the curve (AUC) and C(max) of parent drug, metabolite I and II to the commercial product, indicating that it might give the similar drug efficacy compared to the sibutramine hydrochloride monohydrate-loaded commercial product in rats. Thus, this solid dispersion system would be useful to deliver poorly water-soluble sibutramine base with enhanced bioavailability.

  7. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF

    NARCIS (Netherlands)

    Vergouw, C.G.; Kostelijk, E.H.; Doejaaren, E.; Hompes, P.G.A.; Lambalk, C.B.; Schats, R.

    2012-01-01

    STUDY QUESTION Does the type of medium used to culture fresh and frozenthawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? SUMMARY ANSWER A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean

  8. Determination of specific growth stages of plant cell suspension cultures by monitoring conductivity changes in the medium.

    Science.gov (United States)

    Hahlbrock, K; Ebel, J; Oaks, A; Auden, J; Liersch, M

    1974-03-01

    Conductivity changes in the medium of cultured soybean (Glycine max L.) cells were shown to be strictly correlated with nitrate uptake and growth of the cultures. A continuous record of the conductivity was used as a simple and reliable method of determining specific growth stages and concomitant peaks in the activities of nitrate reductase and phenylalanine ammonia-lyase.

  9. Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp. GO5.

    Science.gov (United States)

    Kim, Mi-Hee; Kong, Yoon-Jung; Baek, Hong; Hyun, Hyung-Hwan

    2006-01-02

    To enhance the production of micrococcin GO5, a bacteriocin produced by Micrococcus sp. GO5, cultivation conditions and medium composition were optimized. The optimal initial pH and temperature for bacteriocin production were 7.0-9.0 and 37 degrees C, respectively. Micrococcus sp. GO5 displayed the highest micrococcin GO5 activity when grown in modified MRS medium that contained lactose or sucrose, rather than glucose, as a carbon source. The maximum bacteriocin activity was obtained in modified MRS medium containing 0.5% tryptone and 1.0% yeast extract as nitrogen sources instead of the other nitrogen sources present in MRS medium. Bacteriocin production was greatly affected by the concentration of K(2)HPO(4); strain GO5 produced eight-fold more bacteriocin in medium containing 2.0-2.5% K(2)HPO(4) than in medium containing 0.2% K(2)HPO(4). The optimal concentration of MgSO(4).7H(2)O for bacteriocin production was 0.5%. The production of micrococcin GO5 was increased 32-fold in shake flask culture and 16-fold in a bioreactor using the optimized medium (TY medium), compared with culturing in MRS medium.

  10. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir ( Pseudotsuga menziesii) shoot cultures

    OpenAIRE

    Chen, Chien-Chih; Bates, Rick; Carlson, John

    2015-01-01

    The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabili...

  11. The Effect of Culture Medium on Metabolic and Antibacterial Activities of Probiotic Bacteria

    Directory of Open Access Journals (Sweden)

    f Mirdavoudi

    2012-05-01

    Full Text Available

    Background and Objectives: Probiotic bacteria is added directly to food components and it has beneficial effect on function and the health of organisms. The bifidogenic factors enter the colon where they contribute to an increase lactic acid bacteria population including Lactobacilli and Bifidobacteria and they inhibit enteric pathogenic bacterial growth. The aim of this study is to investigate the effect of culture medium on metabolic and antibacterial of probiotic bacteria.

     

    Methods: In this study, the probiotics bacterial and intestine pathogenic are to be used. Lactobacilli and Bifidobacterium were identified by plating samples on MRS medium, Gram Staining and standard biochemical methods. The effect of antagonistic probiotics was investigated in the presence of growth factor in the method well diffusion Ager on the Shigella flexneri (PTCC 1234, Escherichia coli (PTCC 1552, Salmonella typhi ( PTCC 1609 and the culture medium pH was measured.

     

    Results: The probiotics bacterial growth in MRS and lactose1%, sorbitol, raffinose, riboflavin were shown the effect antibacterial. The results of the study show the most antagonistic activity in commercial strain Lactobacillus acidophilus on Shigella flexneri and lower activity was in Lactobacillus casei (PTCC 1608, and Salmonella typhimurium (PTCC 1609, and also in Bbifidobacterium bifidum, it showed the most decrease pH value.

     

    Conclusion: According to the result of the study, adding growth factors to MRS medium base and lactose 1%, probiotic growth was increased and which also increased antagonistic activity.

     

  12. Non-invasive optical detection of glucose in cell culture nutrient medium

    Science.gov (United States)

    Cote, Gerald L.

    1993-01-01

    The objective of the proposed research was to begin the development of a non-invasive optical sensor for measuring glucose concentration in the output medium of cell cultures grown in a unique NASA bioreactor referred to as an integrated rotating-wall vessel (IRWV). The input, a bovine serum based nutrient media, has a known glucose concentration. The cells within the bioreactor digest a portion of the glucose. Thus, the non-invasive optical sensor is needed to monitor the decrease in glucose due to cellular consumption since the critical parameters for sustained cellular productivity are glucose and pH. Previous glucose sensing techniques have used chemical reactions to quantify the glucose concentration. Chemical reactions, however, cannot provide for continuous, real time, non-invasive measurement as is required in this application. Our effort while in the fellowship program was focused on the design, optical setup, and testing of one bench top prototype non-invasive optical sensor using a mid-infrared absorption spectroscopy technique. Glucose has a fundamental vibrational absorption peak in the mid-infrared wavelength range at 9.6 micron. Preliminary absorption data using a CO2 laser were collected at this wavelength for water based glucose solutions at different concentrations and one bovine serum based nutrient medium (GTSF) with added glucose. The results showed near linear absorption responses for the glucose-in-water data with resolutions as high at 108 mg/dl and as low as 10 mg/dl. The nutrient medium had a resolution of 291 mg/dl. The variability of the results was due mainly to thermal and polarization drifts of the laser while the decrease in sensitivity to glucose in the nutrient medium was expected due to the increase in the number of confounders present in the nutrient medium. A multispectral approach needs to be used to compensate for these confounders. The CO2 laser used for these studies was wavelength tunable (9.2 to 10.8 micrometers), however

  13. Evaluation of conidia production and mycelial growth in solid culture ...

    African Journals Online (AJOL)

    FATIMAYALBERTO

    2012-10-02

    Oct 2, 2012 ... liquid culture media of different species of ..... Jackson MA, McGuire MR, Lacey LA, Wraight SP (1997). Liquid culture ... fermentation and the substrate in the production of Trichoderma ... Conidiation induction in Penicillium.

  14. Electromechanical and elastic probing of bacteria in a cell culture medium

    International Nuclear Information System (INIS)

    Thompson, G L; Reukov, V V; Vertegel, A A; Nikiforov, M P; Jesse, S; Kalinin, S V

    2012-01-01

    Rapid phenotype characterization and identification of cultured cells, which is needed for progress in tissue engineering and drug testing, requires an experimental technique that measures physical properties of cells with sub-micron resolution. Recently, band excitation piezoresponse force microscopy (BEPFM) has been proven useful for recognition and imaging of bacteria of different types in pure water. Here, the BEPFM method is performed for the first time on physiologically relevant electrolyte media, such as Dulbecco’s phosphate-buffered saline (DPBS) and Dulbecco’s modified Eagle’s medium (DMEM). Distinct electromechanical responses for Micrococcus lysodeikticus (Gram-positive) and Pseudomonas fluorescens (Gram-negative) bacteria in DPBS are demonstrated. The results suggest that mechanical properties of the outer surface coating each bacterium, as well as the electrical double layer around them, are responsible for the BEPFM image formation mechanism in electrolyte media. (paper)

  15. Effect of culture medium volume and embryo density on early mouse embryonic development: tracking the development of the individual embryo.

    Science.gov (United States)

    Dai, Shan-Jun; Xu, Chang-Long; Wang, Jeffrey; Sun, Ying-Pu; Chian, Ri-Cheng

    2012-07-01

    To determine the optimal volume or density of embryos for the well-of-the-well (WOW) system in order to track the development of individual embryos and to determine whether the WOW system can reverse the negative impact of culturing embryos singly. (1) Mouse embryos (groups of nine at the 2-cell stage) were cultured in 6.25 μl, 12.50 μl, 25.00 μl and 50.00 μl of droplets of culture medium under paraffin oil; (2) Groups of three, six, nine and twelve embryos at the 2-cell stage were cultured in 50 μl of droplet of culture medium under paraffin oil; (3) Groups of nine embryos at the 2-cell stage were cultured in 50 μl of droplet under paraffin oil with or without nine micro-wells made on the bottom of the Petri dish into each of which were placed one of the nine embryos (WOW system). Also single 2-cell stage embryos was cultured individually in 5.5 μl of droplet of culture medium under paraffin oil with or without a single micro-well made on the bottom of the Petri dish (WOW system for single culture). At the end of culture, the percentages of blastocyst development, hatching and hatched blastocysts were compared in each group. The blastocysts were fixed for differential staining. The blastocyst development was significantly higher (P WOW system. The blastocyst development was not improved when single embryo cultured individually in a micro-well was compared to single embryo cultured individually without micro-well. The total cell numbers of blastocysts were significantly higher in group embryo culture than single embryo culture regardless of whether the WOW system was used. In addition, the total cell numbers of blastocysts were significantly higher (P WOW system than without. Group embryo culture is superior to single embryo culture for blastocyst development. The WOW system with 50 μl of droplet of culture medium can be used to track the individual development of embryo cultured in groups while preserving good embryonic development. The reduced

  16. Cell culture medium improvement by rigorous shuffling of components using media blending.

    Science.gov (United States)

    Jordan, Martin; Voisard, Damien; Berthoud, Antoine; Tercier, Laetitia; Kleuser, Beate; Baer, Gianni; Broly, Hervé

    2013-01-01

    A novel high-throughput methodology for the simultaneous optimization of many cell culture media components is presented. The method is based on the media blending approach which has several advantages as it works with ready-to-use media. In particular it allows precise pH and osmolarity adjustments and eliminates the need of concentrated stock solutions, a frequent source of serious solubility issues. In addition, media blending easily generates a large number of new compositions providing a remarkable screening tool. However, media blending designs usually do not provide information on distinct factors or components that are causing the desired improvements. This paper addresses this last point by considering the concentration of individual medium components to fix the experimental design and for the interpretation of the results. The extended blending strategy was used to reshuffle the 20 amino acids in one round of experiments. A small set of 10 media was specifically designed to generate a large number of mixtures. 192 mixtures were then prepared by media blending and tested on a recombinant CHO cell line expressing a monoclonal antibody. A wide range of performances (titers and viable cell density) was achieved from the different mixtures with top titers significantly above our previous results seen with this cell line. In addition, information about major effects of key amino acids on cell densities and titers could be extracted from the experimental results. This demonstrates that the extended blending approach is a powerful experimental tool which allows systematic and simultaneous reshuffling of multiple medium components.

  17. Optimization of Culture Medium for Lactobacillus bulgaricus using Box-Behnken Design

    Directory of Open Access Journals (Sweden)

    Zhang Bowen

    2017-06-01

    Full Text Available Lactobacillus bulgaricus is a common yogurt starter in dairy production. But the viable counts of the bacteria in the productions are relatively low during free-drying and storage which is not good for its commercial production. In order to obtain a medium with high activity and high density for bacterial cultured, the experiments and regression analysis were conducted by Box-Behnken design in this study, and a model was established to predict the influence of glucose (9-11 g·L−1, casein hydrolysate (15-17 g·L−1 and glutamate (6.5-7.5 mg·L−1 on viable counts of L. bulgaricus and. The results showed that the glucose, 9.5 g·L−1; casein hydrolysate, 15.5 g·L−1; glutamate, 7.0mg·L−1, the number of viable bacteria of L. bulgaricus could reach (2.95±0.07 ×109, which was very similar to the predicted value of the model of 3.00×109 cfu·mL−1, indicating that the optimized conditions and models used were feasible and effective. The optimized medium components can improve the viable counts of bacteria which are useful from its application in industrial production.

  18. Quantification of the aggregation of magnetic nanoparticles with different polymeric coatings in cell culture medium

    International Nuclear Information System (INIS)

    Eberbeck, D; Zirpel, P; Trahms, L; Kettering, M; Hilger, I; Bergemann, C

    2010-01-01

    The knowledge of the physico-chemical characteristics of magnetic nanoparticles (MNPs) is essential to enhance the efficacy of MNP-based therapeutic treatments (e.g. magnetic heating, magnetic drug targeting). According to the literature, the MNP uptake by cells may depend on the coating of MNPs, the surrounding medium as well as on the aggregation behaviour of the MNPs. Therefore, in this study, the aggregation behaviour of MNPs in various media was investigated. MNPs with different coatings were suspended in cell culture medium (CCM) containing fetal calf serum (FCS) and the distribution of the hydrodynamic sizes was measured by magnetorelaxometry (MRX). FCS as well as bovine serum albumin (BSA) buffer (phosphate buffered saline with 0.1% bovine serum albumin) may induce MNP aggregation. Its strength depends crucially on the type of coating. The degree of aggregation in CCM depends on its FCS content showing a clear, local maximum at FCS concentrations, where the IgG concentration (part of FCS) is of the order of the MNP number concentration. Thus, we attribute the observed aggregation behaviour to the mechanism of agglutination of MNPs by serum compartments as for example IgG. No aggregation was induced for MNPs coated with dextran, polyarabic acid or sodium phosphate, respectively, which were colloidally stable in CCM.

  19. L-Threonine and its analogue added to autoclaved solid medium suppress trichothecene production by Fusarium graminearum.

    Science.gov (United States)

    Maeda, Kazuyuki; Nakajima, Yuichi; Tanahashi, Yoshikazu; Kitou, Yoshiyuki; Miwa, Akihiro; Kanamaru, Kyoko; Kobayashi, Tetsuo; Nishiuchi, Takumi; Kimura, Makoto

    2017-08-01

    Fusarium graminearum produces trichothecene mycotoxins under certain nutritional conditions. When L-Thr and its analogue L-allo-threonine were added to brown rice flour solid medium before inoculation, trichothecene production after 4 days of incubation was suppressed. A time-course analysis of gene expression demonstrated that L-Thr suppressed transcription of Tri6, a trichothecene master regulator gene, and a terpene cyclase Tri5 gene. Regulation of trichothecene biosynthesis by altering major primary metabolic processes may open up the possibility to develop safe chemicals for the reduction of mycotoxin contamination might be developed.

  20. Explosive Breakup of a Water Droplet with a Nontransparent Solid Inclusion Heated in a High-Temperature Gaseous Medium

    Directory of Open Access Journals (Sweden)

    Dmitrienko Margarita A.

    2015-01-01

    Full Text Available This paper investigates the evaporation of a water droplet with a comparably sized solid nontransparent inclusion in a high-temperature (500–800 K gas medium. Water evaporates from the free surface of the inclusion. During this process, intensive vapor formation occurs on the inner interface “water droplet – solid inclusion” with the subsequent explosive decay of the droplet. Experiments have been conducted using high-speed (up to 105 fps video cameras “Phantom” and software “Phantom Camera Control”. The conditions of the explosive vapor formation of the heterogeneous water droplet were found. The typical phase change mechanisms of the heterogeneous water droplet under the conditions of intensive heat exchange were determined.

  1. LED-CT Scan for pH Distribution on a Cross-Section of Cell Culture Medium.

    Science.gov (United States)

    Higashino, Nobuya; Takayama, Toshio; Ito, Hiroaki; Horade, Mitsuhiro; Yamaguchi, Yasutaka; Dylan Tsai, Chia-Hung; Kaneko, Makoto

    2018-01-11

    In cell culture, the pH of the culture medium is one of the most important conditions. However, the culture medium may have non-uniform pH distribution due to activities of cells and changes in the environment. Although it is possible to measure the pH distribution with an existing pH meter using distributed electrodes, the method involves direct contact with the medium and would greatly increase the risk of contamination. Here in this paper, we propose a computed tomography (CT) scan for measuring pH distribution using the color change of phenol red with a light-emitting diode (LED) light source. Using the principle of CT scan, we can measure pH distribution without contacting culture medium, and thus, decrease the risk of contamination. We have developed the device with a LED, an array of photo receivers and a rotation mechanism. The system is firstly calibrated with different shapes of wooden objects that do not pass light, we succeeded in obtaining their 3D topographies. The system was also used for measuring a culture medium with two different pH values, it was possible to obtain a pH distribution that clearly shows the boundary.

  2. Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response.

    Science.gov (United States)

    Haniu, Hisao; Saito, Naoto; Matsuda, Yoshikazu; Tsukahara, Tamotsu; Maruyama, Kayo; Usui, Yuki; Aoki, Kaoru; Takanashi, Seiji; Kobayashi, Shinsuke; Nomura, Hiroki; Okamoto, Masanori; Shimizu, Masayuki; Kato, Hiroyuki

    2013-09-01

    We examined the cytotoxicity of multi-walled carbon nanotubes (MWCNTs) and the resulting cytokine secretion in BEAS-2B cells or normal human bronchial epithelial cells (HBEpCs) in two types of culture media (Ham's F12 containing 10% FBS [Ham's F12] and serum-free growth medium [SFGM]). Cellular uptake of MWCNT was observed by fluorescent microscopy and analyzed using flow cytometry. Moreover, we evaluated whether MWCNT uptake was suppressed by 2 types of endocytosis inhibitors. We found that BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM showed similar biological responses, but BEAS-2B cells cultured in SFGM did not internalize MWCNTs, and the 50% inhibitory concentration value, i.e., the cytotoxicity, was increased by more than 10-fold. MWCNT uptake was suppressed by a clathrin-mediated endocytosis inhibitor and a caveolae-mediated endocytosis inhibitor in BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM. In conclusion, we suggest that BEAS-2B cells cultured in a medium containing serum should be used for the safety evaluation of nanomaterials as a model of normal human bronchial epithelial cells. However, the culture medium composition may affect the proteins that are expressed on the cytoplasmic membrane, which may influence the biological response to MWCNTs. Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.

  3. Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2017-10-01

    Full Text Available Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg2+, Fe2+, and Zn2+ concentrations were adjusted in a minimal mineral (MM medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy.

  4. Medium optimization for nuclease P1 production by Penicillium citrinum in solid-state fermentation using polyurethane foam as inert carrier

    NARCIS (Netherlands)

    Zhu, Y.; Knol, W.; Smits, J.P.; Bol, J.

    1996-01-01

    A solid-state fermentation system, using polyurethane foam as an inert carrier, was used for the production of nuclease P1 by Penicillium citrinum. Optimization of nuclease P1 production was carried out using a synthetic liquid medium. After a two-step medium optimization using a fractional

  5. Ribonucleic artefacts: are some extracellular RNA discoveries driven by cell culture medium components?

    Science.gov (United States)

    Tosar, Juan Pablo; Cayota, Alfonso; Eitan, Erez; Halushka, Marc K; Witwer, Kenneth W

    2017-01-01

    In a recently published study, Anna Krichevsky and colleagues raise the important question of whether results of in vitro extracellular RNA (exRNA) studies, including extracellular vesicle (EV) investigations, are confounded by the presence of RNA in cell culture medium components such as foetal bovine serum (FBS). The answer, according to their data, is a resounding "yes". Even after lengthy ultracentrifugation to remove bovine EVs from FBS, the majority of exRNA in FBS remained. Although technical factors may affect the degree of depletion, residual EVs and exRNA in FBS could influence the conclusions of in vitro studies: certainly, for secreted RNA, and possibly also for cell-associated RNA. In this commentary, we critically examine some of the literature in this field, including a recent study from some of the authors of this piece, in light of the Wei et al. study and explore how cell culture-derived RNAs may affect what we think we know about EV RNAs. These findings hold particular consequence as the field moves towards a deeper understanding of EV-RNA associations and potential functions.

  6. Study on recycling of waste rubbers as medium components for hydroponic culture of rose

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin-Kuk; Lee, Hyung-Gyu; Jeong, Byoung-Ryong; Hwang, Seung-Jae [Gyeongsang National Univ., Kumi(Korea)

    2000-06-30

    Recently, the efficient disposal of the waste rubber is necessary due to increasing amount of the waste rubbers. In this paper, method of recycling waste rubbers as components of medium for hydroponic rose culture was suggested. We investigated growth of rose, and macro- and micro-elements, pH and EC of the media amended with waste rubber. In the beginning of culture, stress symptoms such as thin brittle stem and incipient wilting were observed, but they disappeared in a few weeks. Concentration of Zn{sup 2+} in media at flowering increased in proportion to contents of waste tire in the media. pH of media at flowering were in the range of 5.70 to 6.35. Rose growth in all media, except in waste rock wool mixed with EPDM powder at 9:3 ratio, was normal and equivalent to the control in terms of stem length, number of stems harvested and fresh weight. (author). 10 refs., 5 tabs., 4 figs.

  7. Diverse Requirements for Microglial Survival, Specification, and Function Revealed by Defined-Medium Cultures.

    Science.gov (United States)

    Bohlen, Christopher J; Bennett, F Chris; Tucker, Andrew F; Collins, Hannah Y; Mulinyawe, Sara B; Barres, Ben A

    2017-05-17

    Microglia, the resident macrophages of the CNS, engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol. Using microglial cultures that have never been exposed to serum, we demonstrate a dramatic and lasting change in phagocytic capacity after serum exposure. Finally, we find that mature microglia rapidly lose signature gene expression after isolation, and that this loss can be reversed by engrafting cells back into an intact CNS environment. These data indicate that the specialized gene expression profile of mature microglia requires continuous instructive signaling from the intact CNS. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Culture medium modulates the behaviour of human dental pulp-derived cells: Technical Note

    Directory of Open Access Journals (Sweden)

    S Lopez-Cazaux

    2006-02-01

    Full Text Available In vitro approaches have extensively been developed to study reparative dentinogenesis. While dental pulp is a source of unidentified progenitors able to differentiate into odontoblast-like cells, we investigated the effect of two media; MEM (1.8mM Ca and 1mM Pi and RPMI 1640 (0.8mM Ca and 5mM Pi on the behaviour of human dental pulp cells. Our data indicate that MEM significantly increased cell proliferation and markedly enhanced the proportion of -smooth muscle actin positive cells, which represent a putative source of progenitors able to give rise to odontoblast-like cells. In addition, MEM strongly stimulated alkaline phosphatase activity and was found to induce expression of transcripts encoding dentin sialophosphoprotein, an odontoblastic marker, without affecting that of parathyroid hormone/parathyroid hormone related protein-receptor and osteonectin. In conclusion, these observations demonstrate that not only proliferation but also differentiation into odontoblast-like cells was induced by rich calcium and poor phosphate medium (MEM as compared to RPMI 1640. This study provides important data for the determination of the optimal culture conditions allowing odontoblast-like differentiation in human pulp cell culture.

  9. In vitro plant regeneration of two cucumber (Cucumis sativum L. genotypes: Effects of explant types and culture medium

    Directory of Open Access Journals (Sweden)

    Grozeva Stanislava

    2014-01-01

    Full Text Available The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

  10. The incorporation of low and medium level radioactive wastes (solids and liquids) in cement

    International Nuclear Information System (INIS)

    Palmer, J.D.; Smith, D.L.

    1985-07-01

    Experimentation has shown that high temperatures generated during the setting of ordinary Portland cement/simulant waste mixes can be significantly reduced by the use of a blend of ground granulated blast furnace slag and ordinary Portland cement. Trials on simulated waste showed that blended cement gave improved stability and a reduction in leach rates, and confirmed that the cement-based process can be used for the immobilisation of most types of low and medium level waste. (U.K.)

  11. Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support

    Directory of Open Access Journals (Sweden)

    Anita Muraglia

    2017-11-01

    Full Text Available Present cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i an heparin-free human platelet lysate (PL devoid of serum or plasma components (v-PL and (ii an heparin-free human serum derived from plasma devoid of PL components (Pl-s and to their use as single components or in combination in primary or cell line cultures. Human mesenchymal stem cells (MSC primary cultures were obtained from adipose tissue, bone marrow, and umbilical cord. Human chondrocytes were obtained from articular cartilage biopsies. In general, MSC expanded in the presence of Pl-s alone showed a low or no proliferation in comparison to cells grown with the combination of Pl-s and v-PL. Confluent, growth-arrested cells, either human MSC or human articular chondrocytes, treated with v-PL resumed proliferation, whereas control cultures, not supplemented with v-PL, remained quiescent and did not proliferate. Interestingly, signal transduction pathways distinctive of proliferation were activated also in cells treated with v-PL in the absence of serum, when cell proliferation did not occur, indicating that v-PL could induce the cell re-entry in the cell cycle (cell commitment, but the presence of serum proteins was an absolute requirement for cell proliferation to happen. Indeed, Pl-s alone supported cell growth in constitutively activated cell lines (U-937, HeLa, HaCaT, and V-79 regardless of the co-presence of v-PL. Plasma- and plasma-derived serum were equally able to sustain cell proliferation although, for cells cultured in adhesion, the Pl-s was more efficient than the plasma from which it was derived. In conclusion, the cells expanded in the presence of the new additives maintained their differentiation potential and did not show alterations in their karyotype.

  12. Dependence of synchronized bursting activity on medium stirring and the perfusion rate in a cultured network of neurons

    Science.gov (United States)

    Heo, Ryoun; Kim, Hyun; Lee, Kyoung J.

    2016-05-01

    A cultured network of neurons coupled with a multi-electrode-array (MEA) recording system has been a useful platform for investigating various issues in neuroscience and engineering. The neural activity supported by the system can be sensitive to environmental fluctuations, for example, in the medium's nutrient composition, ph, and temperature, and to mechanical disturbances, yet this issue has not been the subject. Especially, a normal practice in maintaining neuronal cell cultures involves an intermittent sequence of medium exchanges, typically at a time interval of a few days, and one such sudden medium exchange is unavoidably accompanied by many unintended disturbances. Here, based on a quantitative time-series analysis of synchronized bursting events, we explicitly demonstrate that such a medium exchange can, indeed, bring a huge change in the existing neural activity. Subsequently, we develop a medium perfusion-stirring system and an ideal protocol that can be used in conjunction with a MEA recording system, providing long-term stability. Specifically, we systematically evaluate the effects of medium stirring and perfusion rates. Unexpectedly, even some vigorous mechanical agitations do not have any impacts on neural activity. On the other hand, too much replenishment ( e.g., 1.8 ml/day for a 1.8-ml dish) of neurobasal medium results in an excitotoxicity.

  13. Solid Waste from Four-stroke Medium Speed Engine Power Plant Operation

    OpenAIRE

    Smart, Heidi

    2016-01-01

    This thesis is a study of the solid-state waste generated in power plants based on internal combustion engine (ICE) technology, supplied by the Wärtsilä Energy Solutions division. This thesis, which characterizes and quantifies the waste, was done to enhance the ability of supporting customers in issues related to social and environmental impact assessment, environmental permits, waste management planning, etc. The Performance Standards on Environmental and Social Sustainability and the E...

  14. Advances in medium and high temperature solid oxide fuel cell technology

    CERN Document Server

    Salvatore, Aricò

    2017-01-01

    In this book well-known experts highlight cutting-edge research priorities and discuss the state of the art in the field of solid oxide fuel cells giving an update on specific subjects such as protonic conductors, interconnects, electrocatalytic and catalytic processes and modelling approaches. Fundamentals and advances in this field are illustrated to help young researchers address issues in the characterization of materials and in the analysis of processes, not often tackled in scholarly books.

  15. Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis.

    Science.gov (United States)

    Kashyap, R S; Ramteke, S S; Gaherwar, H M; Deshpande, P S; Purohit, H J; Taori, G M; Daginawala, H

    2010-01-01

    The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.

  16. Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Kashyap R

    2010-01-01

    Full Text Available The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth for the diagnosis of tuberculous meningitis (TBM in cerebrospinal fluid (CSF. CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50 were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64% out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI: 67-93%] and 36% specificity (95% CI: 57-98% for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM. This coexistence may go undetected or potentially lead to erroneous reporting of results.

  17. Implications in studies of environmental risk assessments: Does culture medium influence the results of toxicity tests of marine bacteria?

    Science.gov (United States)

    Díaz-García, Alejandra; Borrero-Santiago, Ana R; Riba, Inmaculada

    2018-04-14

    Two marine bacterial populations (Roseobacter sp. and Pseudomonas litoralis) were exposed to different concentrations of zinc (300, 625, 1250, 2000, 2500 and 5000 mg L -1 ) and cadmium (75, 250, 340, 500 and 1000 mg L -1 ) using two culture media (full nutrient Marine Broth 2216 "MB" and 1:10 (vol/vol) dilution with seawater of Marine Broth 2216 "MB SW "), in order to assess population responses depending on the culture medium and also potential adverse effects associated with these two metals. Different responses were found depending on the culture medium (Bacterial abundance (cells·mL -1 ), growth rates (μ, hours -1 ), and production of Extracellular Polysaccharides Substances (EPS) (μg glucose·cells -1 ). Results showed negative effects in both strains after the exposure to Zn treatments. Both strains showed highest metal sensitivity at low concentrations using both culture media. However, different results were found when exposing the bacterial populations to Cd treatments depending on the culture medium. Highest toxicity was observed using MB at low levels of Cd concentrations, whereas MB SW showed toxicity to bacteria at higher concentrations of Cd. Results not only showed adverse effects on Roseobacter sp. and Pseudomonas litoralis associated with the concentration of Zn and Cd, but also confirm that depending on the culture medium results can differ. This work suggests MB SW as an adequate culture medium to study metal toxicity bioassays in order to predict realistic effects on marine bacterial populations. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Medium from X-rayed cultures induces DNA strand-breaks in non-irradiated HeLa cells

    International Nuclear Information System (INIS)

    Ikushima, T.; Okuyama, K.; Tanizaki, Y.

    2002-01-01

    There is growing evidence to indicate that several types of responses are induced by ionizing radiation in non-irradiated cells. Such bystander effects include the killing of non-irradiated cells, the induction of sister chromatid exchanges and chromosomal aberrations, and the induction of gene mutations and chromosomal instability and enhanced cell growth. In the present study, we assessed whether the medium from irradiated cultures can induce DNA strand-breaks in non-irradiated cells, using single-cell gel electrophoresis assay (comet assay). HeLa cells in culture were irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken from the irradiated culture, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-target cells. After incubation for 30 min, the comet assay was performed under alkaline and neutral conditions. Such treatments resulted in a dose-dependent increase in tail moment under either alkaline or neutral condition, indicating the induction of DNA single- or double-strand breaks, respectively. It was also shown that the clonogenic survival was reduced in the cells cultured in the medium from irradiated cultures. Such a change was not detected at all when medium alone was irradiated. These results provided disputed evidence that irradiated cells released certain genotoxic factor(s) into the culture medium that can induce DNA strand breaks leading to cell death. Our results suggest that physical contact between irradiated and non-irradiated cells may not be necessary for the bystander effects observed in this study. It appears that bystander responses may be mediated by multiple mechanisms

  19. Deriving a Planting Medium from Solid Waste Compost and Construction, Demolition and Excavation Waste

    Science.gov (United States)

    Farajalla, Nadim; Assaf, Eleni; Bashour, Issam; Talhouk, Salma

    2014-05-01

    Lebanon's very high population density has been increasing since the end of the war in the early 1990s reaching 416.36 people per square kilometer. Furthermore, the influx of refugees from conflicts in the region has increased the resident population significantly. All these are exerting pressure on the country's natural resources, pushing the Lebanese to convert more forest and agricultural land into roads, buildings and houses. This has led to a building boom and rapid urbanization which in turn has created a demand for construction material - mainly rock, gravel, sand, etc. nearly all of which were locally acquired through quarrying to the tune of three million cubic meters annually. This boom has been followed by a war with Israel in 2006 which resulted in thousands of tonnes of debris. The increase in population has also led to an increase in solid waste generation with 1.57 million tonnes of solid waste generated in Lebanon per year. The combination of construction, demolition and excavation (CDE) waste along with the increase in solid waste generation has put a major stress on the country and on the management of its solid waste problem. Compounding this problem are the issues of quarries closure and rehabilitation and a decrease in forest and vegetative cover. The on-going research reported in this paper aims to provide an integrated solution to the stated problem by developing a "soil mix" derived from a mélange of the organic matter of the solid waste (compost), the CDE waste, and soil. In this mix, native and indicator plants are planted (in pots) from which the most productive mix will be selected for further testing at field level in later experiments. The plant species used are Matiolla, a native Lebanese plant and Zea mays, which is commonly known used as an indicator plant due to its sensitivity to environmental conditions. To ensure sustainability and environmental friendliness of the mix, its physical and chemical characteristics are monitored

  20. Solid-stemmed spring wheat cultivars give better androgenic response than hollow-stemmed cultivars in anther culture

    OpenAIRE

    Weigt, Dorota; Kiel, Angelika; Nawraca?a, Jerzy; Pluta, Mateusz; ?acka, Agnieszka

    2016-01-01

    Solid-stemmed spring wheat cultivars (Triticum aestivum L.) are resistant to the stem sawfly (Cephus cinctus Nort.) and lodging. Anthers of 24 spring wheat cultivars with varying content of pith in the stem were used in the experiment. All were classified into three groups: solid, medium?solid and hollow stems. There was considerable influence of the cultivar on callus formation and green plant regeneration. The highest efficiency of green plant regeneration (24%) was observed for the solid-s...

  1. Age of G-1 PLUS v5 embryo culture medium is inversely associated with birthweight of the newborn.

    Science.gov (United States)

    Kleijkers, Sander H M; van Montfoort, Aafke P A; Smits, Luc J M; Coonen, Edith; Derhaag, Josien G; Evers, Johannes L H; Dumoulin, John C M

    2015-06-01

    Does age of G-1 PLUS v5 embryo culture medium affect IVF outcome? Birthweight of singletons born after IVF showed an inverse association with age of the embryo culture medium, while no association was found between age of culture medium and fertilization rate, embryonic development or ongoing pregnancy. It has been reported that IVF culture media can deteriorate during storage, which suggests that the capacity of culture media to support optimal embryo development decreases over time. Some animal studies showed an effect of storage time on embryo development, in contrast to other studies, while the effect of aging culture medium on IVF outcome in humans is unknown. We used data on outcome of 1832 IVF/ICSI cycles with fresh embryo transfer, performed in the period 2008-2012 to evaluate the association of fertilization rate, embryonic development, ongoing pregnancy and birthweight of singletons with age of the culture medium (Vitrolife AB G-1 PLUS v5). Age of the culture medium was calculated by subtracting the production date from the date of ovum retrieval. Data analysis included linear regression and logistic regression on continuous and categorical outcomes, respectively. Age of the culture medium was not associated with fertilization rate (P = 0.543), early cleavage rate (P = 0.155), percentage of embryos containing four or more cells on Day 2 (P = 0.401), percentage of embryos containing eight or more cells on Day 3 (P = 0.175), percentage of embryos with multinucleated blastomeres (P = 0.527), or ongoing pregnancy (P = 0.729). However, birthweight of the newborn was inversely associated with age of the medium (β = -3.6 g, SE: 1.5 g, P = 0.021), after controlling for possible confounders (day of embryo transfer, number of transferred embryos, child's gender, gestational age at birth, parity, pregnancy complications, maternal smoking, height and weight, and paternal height and weight) and the association was not biased by year of treatment, time since first

  2. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF.

    Science.gov (United States)

    Vergouw, Carlijn G; Kostelijk, E Hanna; Doejaaren, Els; Hompes, Peter G A; Lambalk, Cornelis B; Schats, Roel

    2012-09-01

    Does the type of medium used to culture fresh and frozen-thawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean birthweight adjusted for gestational age, gender and parity (z-scores) of singletons born after a fresh or frozen-thawed SET. Furthermore, we show that embryo freezing and thawing cycles may lead to a significantly higher mean birthweight. Animal studies have shown that culture media constituents are responsible for changes in birthweight of offspring. In human IVF, there is still little knowledge of the effect of medium type on birthweight. Until now, only a small number of commercially available culture media have been investigated (Vitrolife, Cook(®) Medical and IVF online medium). Our study adds new information: it has a larger population of singleton births compared with the previously published studies, it includes outcomes of other media types (HTF and Sage(®)), not previously analysed, and it includes data on frozen-thawed SETs. This study was a retrospective analysis of birthweights of singleton newborns after fresh (Day 3) or frozen-thawed (Day 5) SET cycles, using embryos cultured in either of two different types of commercially available culture media, between 2008 and 2011. Before January 2009, a single-step culture medium was used: human tubal fluid (HTF) with 4 mg/ml human serum albumin. From January 2009 onwards, a commercially available sequential medium was introduced: Sage(®), Quinn's advantage protein plus medium. Singletons born after a fresh SET (99 embryos cultured in HTF and 259 in Sage(®)) and singletons born after a frozen-thawed SET (32 embryos cultured in HTF only, 41 in HTF and Sage(®) and 86 in Sage(®) only) were analysed. Only patients using autologous gametes without the use of a gestational carrier were considered. Also excluded were (vanishing) twins, triplets

  3. Increasing efficiency of human mesenchymal stromal cell culture by optimization of microcarrier concentration and design of medium feed.

    Science.gov (United States)

    Chen, Allen Kuan-Liang; Chew, Yi Kong; Tan, Hong Yu; Reuveny, Shaul; Weng Oh, Steve Kah

    2015-02-01

    Large amounts of human mesenchymal stromal cells (MSCs) are needed for clinical cellular therapy. In a previous publication, we described a microcarrier-based process for expansion of MSCs. The present study optimized this process by selecting suitable basal media, microcarrier concentration and feeding regime to achieve higher cell yields and more efficient medium utilization. MSCs were expanded in stirred cultures on Cytodex 3 microcarriers with media containing 10% fetal bovine serum. Process optimization was carried out in spinner flasks. A 2-L bioreactor with an automated feeding system was used to validate the optimized parameters explored in spinner flask cultures. Minimum essential medium-α-based medium supported faster MSC growth on microcarriers than did Dulbecco's modified Eagle's medium (doubling time, 31.6 ± 1.4 vs 42 ± 1.7 h) and shortened the process time. At microcarrier concentration of 8 mg/mL, a high cell concentration of 1.08 × 10(6) cells/mL with confluent cell concentration of 4.7 × 10(4)cells/cm(2) was achieved. Instead of 50% medium exchange every 2 days, we have designed a full medium feed that is based on glucose consumption rate. The optimal medium feed that consisted of 1.5 g/L glucose supported MSC growth to full confluency while achieving the low medium usage efficiency of 3.29 mL/10(6)cells. Finally, a controlled bioreactor with the optimized parameters achieved maximal confluent cell concentration with 16-fold expansion and a further improved medium usage efficiency of 1.68 mL/10(6)cells. We have optimized the microcarrier-based platform for expansion of MSCs that generated high cell yields in a more efficient and cost-effective manner. This study highlighted the critical parameters in the optimization of MSC production process. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  4. Solid waste from leather industry as adsorbent of organic dyes in aqueous-medium

    International Nuclear Information System (INIS)

    Oliveira, Luiz C.A.; Goncalves, Maraisa; Oliveira, Diana Q.L.; Guerreiro, Mario C.; Guilherme, Luiz R.G.; Dallago, Rogerio M.

    2007-01-01

    The industrial tanning of leather usually produces considerable amounts of chromium-containing solid waste and liquid effluents and raises many concerns on its environmental effect as well as on escalating landfill costs. Actually, these shortcomings are becoming increasingly a limiting factor to this industrial activity that claims for alternative methods of residue disposals. In this work, it is proposed a novel alternative destination of the solid waste, based on the removal of organic contaminants from the out coming aqueous-residue. The adsorption isotherm pattern for the wet blue leather from the Aurea tanning industry in Erechim-RS (Brazil) showed that these materials present high activity on adsorbing the reactive red textile dye as well as other compounds. The adsorbent materials were characterized by IR spectroscopy and SEM and tested for the dye adsorption (reactive textile and methylene blue dyes). The concentrations of dyes were measured by UV-vis spectrophotometry and the chromium extraction from leather waste was realized by basic hydrolysis and determined by atomic absorption. As a low cost abundant adsorbent material with high adsorption ability on removing dye methylene blue (80 mg g -1 ) and textile dye reactive red (163 mg g -1 ), the leather waste is revealed to be a interesting alternative relatively to more costly adsorbent materials

  5. Solid waste from leather industry as adsorbent of organic dyes in aqueous-medium

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Luiz C.A. [Universidade Federal de Lavras, Depto. de Quimica, Caixa Postal 37, CEP 37200.000, Lavras-MG (Brazil)]. E-mail: luizoliveira@ufla.br; Goncalves, Maraisa [Universidade Federal de Lavras, Depto. de Quimica, Caixa Postal 37, CEP 37200.000, Lavras-MG (Brazil); Oliveira, Diana Q.L. [Universidade Federal de Lavras, Depto. de Quimica, Caixa Postal 37, CEP 37200.000, Lavras-MG (Brazil); Guerreiro, Mario C. [Universidade Federal de Lavras, Depto. de Quimica, Caixa Postal 37, CEP 37200.000, Lavras-MG (Brazil); Guilherme, Luiz R.G. [Universidade Federal de Lavras, Depto. de Ciencia do solo, CEP 37200.000, Lavras-MG (Brazil); Dallago, Rogerio M. [URI-Campus Erechim, Av. 7 Setembro 1621, Centro, CEP 99700-000, Depto de Quimica, Erechim-RS (Brazil)

    2007-03-06

    The industrial tanning of leather usually produces considerable amounts of chromium-containing solid waste and liquid effluents and raises many concerns on its environmental effect as well as on escalating landfill costs. Actually, these shortcomings are becoming increasingly a limiting factor to this industrial activity that claims for alternative methods of residue disposals. In this work, it is proposed a novel alternative destination of the solid waste, based on the removal of organic contaminants from the out coming aqueous-residue. The adsorption isotherm pattern for the wet blue leather from the Aurea tanning industry in Erechim-RS (Brazil) showed that these materials present high activity on adsorbing the reactive red textile dye as well as other compounds. The adsorbent materials were characterized by IR spectroscopy and SEM and tested for the dye adsorption (reactive textile and methylene blue dyes). The concentrations of dyes were measured by UV-vis spectrophotometry and the chromium extraction from leather waste was realized by basic hydrolysis and determined by atomic absorption. As a low cost abundant adsorbent material with high adsorption ability on removing dye methylene blue (80 mg g{sup -1}) and textile dye reactive red (163 mg g{sup -1}), the leather waste is revealed to be a interesting alternative relatively to more costly adsorbent materials.

  6. Organisational culture as a part in the development of open innovation - the perspective of small and medium-sized enterprises

    Directory of Open Access Journals (Sweden)

    Szymańska Katarzyna

    2016-05-01

    Full Text Available The ability to introduce various concepts and business models is nowadays a prerequisite of creating a competitive advantage. This is to a large extent closely linked to the ability of enterprises to create, implement and disseminate a variety of innovative solutions. Today the use of open innovation is a necessity. This applies not only to large organisations, but also to small and medium-sized enterprises. In order to implement open innovation, small and medium-sized enterprises need to effectively manage their own growth through the preparation of appropriate strategies and the development of a model that encompasses all changes, taking into account a number of factors related to the growth dynamics of this sector. It is understood that an appropriate organisational culture plays an important role in the implementation of innovation in the sector of small and medium-sized enterprises. There are many indications that a cultural mismatch and misunderstanding are the main reasons for major problems related to the low level of implementation of innovation by small and medium-sized enterprises. The aim of the paper is to outline the issue of the impact of organisational culture on the development of the concept of open innovation in the sector of small and medium-sized enterprises.

  7. Solid Culturing of Bacillus amyloliquefaciens for α-Amylase Production

    Directory of Open Access Journals (Sweden)

    Dhanya Gangadharan

    2006-01-01

    Full Text Available Fourteen different agroresidues were screened for alpha amylase production using Bacillus amyloliquefaciens ATCC 23842. Among them, wheat bran (WB and groundnut oil cake (GOC in mass ratio of 1:1 was proved as the best substrate source. Supplementation with 0.01 M KH2PO4 and 1 % soluble starch enhanced the enzyme yield considerably. Maximum enzyme recovery from the solid mass was obtained when extracted with 0.1 M acetate buffer, pH=5.0. Maximum enzyme titer expressed as units per mass of dry substrate obtained was 62 470 U/g after 72 hours of fermentation at 37 °C by using the above solid substrate mixture (5 g with the initial moisture of 85 % and inoculated with Bacillus amyloliquefaciens of 2·109 CFU/mL.

  8. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir (Pseudotsuga menziesii shoot cultures [version 2; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Chien-Chih Chen

    2015-05-01

    Full Text Available The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1 determine medium pH change over time in storage conditions and with presence of explants, 2 evaluate the effects of medium pH change on explant growth performance and 3 assess the effects of adding a pH stabilizer, 2-(N-morpholinoethanesulfonic acid (MES that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth.

  9. Agglomeration, sedimentation, and cellular toxicity of alumina nanoparticles in cell culture medium

    International Nuclear Information System (INIS)

    Yoon, Dokyung; Woo, Daekwang; Kim, Jung Heon; Kim, Moon Ki; Kim, Taesung; Hwang, Eung-Soo; Baik, Seunghyun

    2011-01-01

    The cytotoxicity of alumina nanoparticles (NPs) was investigated for a wide range of concentration (25–200 μg/mL) and incubation time (0–72 h) using floating cells (THP-1) and adherent cells (J774A.1, A549, and 293). Alumina NPs were gradually agglomerated over time although a significant portion of sedimentation occurred at the early stage within 6 h. A decrease of the viability was found in floating (THP-1) and adherent (J774A.1 and A549) cells in a dose-dependent manner. However, the time-dependent decrease in cell viability was observed only in adherent cells (J774A.1 and A549), which is predominantly related with the sedimentation of alumina NPs in cell culture medium. The uptake of alumina NPs in macrophages and an increased cell-to-cell adhesion in adherent cells were observed. There was no significant change in the viability of 293 cells. This in vitro test suggests that the agglomeration and sedimentation of alumina NPs affected cellular viability depending on cell types such as monocytes (THP-1), macrophages (J774A.1), lung carcinoma cells (A549), and embryonic kidney cells (293).

  10. Agglomeration, sedimentation, and cellular toxicity of alumina nanoparticles in cell culture medium

    Science.gov (United States)

    Yoon, Dokyung; Woo, Daekwang; Kim, Jung Heon; Kim, Moon Ki; Kim, Taesung; Hwang, Eung-Soo; Baik, Seunghyun

    2011-06-01

    The cytotoxicity of alumina nanoparticles (NPs) was investigated for a wide range of concentration (25-200 μg/mL) and incubation time (0-72 h) using floating cells (THP-1) and adherent cells (J774A.1, A549, and 293). Alumina NPs were gradually agglomerated over time although a significant portion of sedimentation occurred at the early stage within 6 h. A decrease of the viability was found in floating (THP-1) and adherent (J774A.1 and A549) cells in a dose-dependent manner. However, the time-dependent decrease in cell viability was observed only in adherent cells (J774A.1 and A549), which is predominantly related with the sedimentation of alumina NPs in cell culture medium. The uptake of alumina NPs in macrophages and an increased cell-to-cell adhesion in adherent cells were observed. There was no significant change in the viability of 293 cells. This in vitro test suggests that the agglomeration and sedimentation of alumina NPs affected cellular viability depending on cell types such as monocytes (THP-1), macrophages (J774A.1), lung carcinoma cells (A549), and embryonic kidney cells (293).

  11. Children’s picturebook on sexual educationas a cultural and political medium

    Directory of Open Access Journals (Sweden)

    Małgorzata Cackowska

    2011-11-01

    Full Text Available In my article I deal with a social construction of meanings of picturebooks’ content and form in Poland and abroad, so also with what kinds of discourses and ideologies determine the conditions of picturebooks’ production in societies under analysis. For the analysis I have chosen picturebooks which deal with sexual education. The methodology applied in the research consists mostly of content analysis and critical discourse analysis. The research is a part of a bigger collaborative project called “Discursive construction of subjectivity” financed by Ministry of Science and Higher Education in Poland, grant no. N 10702632/3637, and conducted at the University of Gdansk. I present, basing on the empirical material, the critique of the dominant discourse in Poland which is powerful in the production of picturebooks, which is based on the conservative ideology and social and sexual roles defined in stereotypical, hierarchical and heterosexual terms. In this aura discourses based on liberal or radical ideologies are marginalized.The results show the knowledge/power relations, symptoms of symbolic violence in exemplified discourses and explain to what practices of ideological and political control the subject is exposed. In this context a picturebook is seen as a meaningful cultural and political medium, within the content and form of which various (possible ideologies and conceptions of the child are included to or excluded from social environment, what can occur as a real issue for educational theory and practice.

  12. Prototypical antipsychotic drugs protect hippocampal neuronal cultures against cell death induced by growth medium deprivation

    Directory of Open Access Journals (Sweden)

    Williams Sylvain

    2006-03-01

    Full Text Available Abstract Background Several clinical studies suggested that antipsychotic-based medications could ameliorate cognitive functions impaired in certain schizophrenic patients. Accordingly, we investigated the effects of various dopaminergic receptor antagonists – including atypical antipsychotics that are prescribed for the treatment of schizophrenia – in a model of toxicity using cultured hippocampal neurons, the hippocampus being a region of particular relevance to cognition. Results Hippocampal cell death induced by deprivation of growth medium constituents was strongly blocked by drugs including antipsychotics (10-10-10-6 M that display nM affinities for D2 and/or D4 receptors (clozapine, haloperidol, (±-sulpiride, domperidone, clozapine, risperidone, chlorpromazine, (+-butaclamol and L-741,742. These effects were shared by some caspases inhibitors and were not accompanied by inhibition of reactive oxygen species. In contrast, (--raclopride and remoxipride, two drugs that preferentially bind D2 over D4 receptors were ineffective, as well as the selective D3 receptor antagonist U 99194. Interestingly, (--raclopride (10-6 M was able to block the neuroprotective effect of the atypical antipsychotic clozapine (10-6 M. Conclusion Taken together, these data suggest that D2-like receptors, particularly the D4 subtype, mediate the neuroprotective effects of antipsychotic drugs possibly through a ROS-independent, caspase-dependent mechanism.

  13. Radiobiological application of atomic force microscopy. Analysis on human chromosomes in culture medium

    International Nuclear Information System (INIS)

    Watanabe, Makoto; Kinjo, Yasuhito

    1995-01-01

    We have proposed a 'Heterogeneous Chromatin Target Model' on the regulating mechanisms involved in chromosome mutation due to ionizing radiations. The heterogeneity of chromatin is derived from the highly condensed organization of chromatin segments that consist of hypersensitive and fragile sites in the fluctuating assembly of nucleosome clusters (superbeads). The above consideration is going to be subjected to a new experimental approach applying the atomic force microscope (AFM), one of the most promising members of a family of scanning probe microscope (SPM). The AFM can be operated in liquid as well as in air. A living specimen can be examined without any preparative procedures (for instance, fixation, staining, vecuum evaporation and so on). Micromanipulation of the isolated chromosome is also possible by the precise positional control of a cantilever on the nanometer scale. In the present report, the mitotic metaphase chromosomes released from living cells (human lymphocytes RPMI) were spread on the clean surface of distilled water filled in a trough. The spread surface film, in which the chromosomes were embedded, was picked up and adhered tightly on a specimen substrate made of silicon. The whole-mounted chromosome were submerged in a solution of culture medium and observed within a liquid immersion cell for AFM. We used an AFM system, SPA-300 made by Seiko Instruments. The particulate chromatin segments of nucleosome clusters (superbeads) were clearly observed within mitotic human chromosomes in a living hydrated condition. These findings support the heterogeneity of chromatin target in a living cell. (author)

  14. Optimization of a Culture Medium Using the Taguchi Approach for the Production of Microorganisms Active in Odorous Compound Removal

    Directory of Open Access Journals (Sweden)

    Krzysztof Makowski

    2017-07-01

    Full Text Available The aim of this work was to develop the composition of a medium for the cultivation of six microbial strains forming a deodorizing consortium: Pseudomonas fluorescens, Enterococcus faecium, Bacillus subtilis, Bacillus megaterium, Leuconostoc mesenteroides and Lactobacillus plantarum. The study focused on the optimization of a highly efficient culture medium composed of readily available components of plant origin to maximize microbial biomass yields, and to create a less expensive alternative to the commercial Tryptic Soy Broth medium (TSB. After preliminary efficiency screening of all tested media components, we selected four substrates for further optimization—soy protein concentrate (SPC, glucose or sucrose, and phosphate salts. The final concentrations of all components were fine-tuned using the Taguchi design for experiments according to an L9 array. Taguchi optimization led to formulation of a culture medium, which was approximately 5 times cheaper than TSB (depending on the components used. Consequently, microbial biomass yields were improved by up to 15-fold (1564%, depending on the strain. The results obtained in the laboratory experiments were then confirmed in pilot- (42 L and industrial- (300 L scale fermentation. Our results show that this method of using a parallel culture microbioreactor with the Taguchi approach can be recommended for optimization of culture media based on substrates of plant origin.

  15. Enhancement of excision-repair efficiency by conditioned medium from density-inhibited cultures in V79 Chinese hamster cells

    International Nuclear Information System (INIS)

    Nakano, S.

    1979-01-01

    Conditioned medium from density-inhibited V79 Chinese hamster cell cultures, given as a post-treatment to UV-irradiated homologous cells, was demonstrated to reduce the lethal action of ultraviolet light by temporarily blocking DNA replication. Since the increased survival was not affected by various nontoxic concentrations of caffeine, such protective effect would be attributable to the prolonged intervention of excision repair before DNA replication during the post-treatment period. The influence of conditioned medium on the UV-induced mutation at the ouabain-resistance locus was also examined and a significant decrease in mutation frequecy was noted. The observed reduction in killing and mutation as a result of post-incubation in conditioned medium, which delays DNA replication, would be interpreted as evidence that conditioned medium provides a longer period of time for an error-free excision-repair process, leaving lesion in DNA available for error-prone post-replication repair. (Auth.)

  16. IVF culture medium affects human intrauterine growth as early as the second trimester of pregnancy.

    Science.gov (United States)

    Nelissen, Ewka C M; Van Montfoort, Aafke P A; Smits, Luc J M; Menheere, Paul P C A; Evers, Johannes L H; Coonen, Edith; Derhaag, Josien G; Peeters, Louis L; Coumans, Audrey B; Dumoulin, John C M

    2013-08-01

    When does a difference in human intrauterine growth of singletons conceived after IVF and embryo culture in two different culture media appear? Differences in fetal development after culture of embryos in one of two IVF media were apparent as early as the second trimester of pregnancy. Abnormal fetal growth patterns are a major risk factor for the development of chronic diseases in adult life. Previously, we have shown that the medium used for culturing embryos during the first few days after fertilization significantly affects the birthweight of the resulting human singletons. The exact onset of this growth difference was unknown. In this retrospective cohort study, all 294 singleton live births after fresh embryo transfer in the period July 2003 to December 2006 were included. These embryos originated from IVF treatments that were part of a previously described clinical trial. Embryos were allocated to culture in either Vitrolife or Cook commercially available sequential culture media. We analysed ultrasound examinations at 8 (n = 290), 12 (n = 83) and 20 weeks' (n = 206) gestation and used first-trimester serum markers [pregnancy-associated plasma protein-A (PAPP-A) and free β-hCG]. Differences between study groups were tested by the Student's t-test, χ(2) test or Fisher's exact test, and linear multivariable regression analysis to adjust for possible confounders (for example, parity, gestational age at the time of ultrasound and fetal gender). A total of 294 singleton pregnancies (Vitrolife group nVL = 168, Cook group: nC = 126) from 294 couples were included. At 8 weeks' gestation, there was no difference between crown-rump length-based and ovum retrieval-based gestational age (ΔGA) (nVL = 163, nC = 122, adjusted mean difference, -0.04 days, P = 0.84). A total of 83 women underwent first-trimester screening at 12 weeks' gestation (nVL = 45, nC = 38). ΔGA, nuchal translucency (multiples of median, MoM) and PAPP-A (MoM) did not differ between the study

  17. Culture medium, gas atmosphere and MAPK inhibition affect regulation of RNA-binding protein targets during mouse preimplantation development.

    Science.gov (United States)

    Calder, Michele D; Watson, Patricia H; Watson, Andrew J

    2011-11-01

    During oogenesis, mammalian oocytes accumulate maternal mRNAs that support the embryo until embryonic genome activation. RNA-binding proteins (RBP) may regulate the stability and turnover of maternal and embryonic mRNAs. We hypothesised that varying embryo culture conditions, such as culture medium, oxygen tension and MAPK inhibition, affects regulation of RBPs and their targets during preimplantation development. STAU1, ELAVL1, KHSRP and ZFP36 proteins and mRNAs were detected throughout mouse preimplantation development, whereas Elavl2 mRNA decreased after the two-cell stage. Potential target mRNAs of RBP regulation, Gclc, Slc2a1 and Slc7a1 were detected during mouse preimplantation development. Gclc mRNA was significantly elevated in embryos cultured in Whitten's medium compared with embryos cultured in KSOMaa, and Gclc mRNA was elevated under high-oxygen conditions. Inhibition of the p38 MAPK pathway reduced Slc7a1 mRNA expression while inhibition of ERK increased Slc2a1 mRNA expression. The half-lives of the potential RBP mRNA targets are not regulated in parallel; Slc2a1 mRNA displayed the longest half-life. Our results indicate that mRNAs and proteins encoding five RBPs are present during preimplantation development and more importantly, demonstrate that expression of RBP target mRNAs are regulated by culture medium, gas atmosphere and MAPK pathways.

  18. Effects of medium components and culture conditions on mycelial biomass and the production of bioactive ingredients in submerged culture of Xylaria nigripes (Ascomycetes), a Chinese medicinal fungus.

    Science.gov (United States)

    Chen, Jian-Zhi; Lo, Hui-Chen; Lin, Fang-Yi; Chang, Shih-Liang; Hsieh, Changwei; Liang, Zeng-Chin; Ho, Wai-Jane; Hsu, Tai-Hao

    2014-01-01

    The optimal culture conditions were investigated to maximize the production of mycelial biomass and bioactive ingredients in submerged cultivation of Xylaria nigripes, a Chinese medicinal fungus. The one-factor-at-a-time method was used to explore the effects of medium components, including carbon, nitrogen, mineral sources, and initial pH of the medium and environmental factors, such as culture temperature and rotation speed, on mycelial growth and production of bioactive ingredients. The results indicated that the optimal culture temperature and rotation speed were 25°C and 100 rpm in a medium with 20 g fructose, 6 g yeast extract, and 2 g magnesiun sulfate heptahydrate as carbon, nitrogen, and mineral sources, respectively, in 1 L distilled water with an initial medium pH of 5.5. With optimal medium components and conditions of cultivation, the maximal production of mycelial biomass was 6.64 ± 0.88 g/L, with maximal production of bioactive ingredients such as extracellular polysaccharides (2.36 ± 0.18 mg/mL), intracellular polysaccharides (2.38 ± 0.07 mg/g), adenosine (43.27 ± 2.37 mg/g), total polyphenols (36.57 ± 1.36 mg/g), and triterpenoids (31.29 ± 1.17 mg/g) in a shake flask culture. These results suggest that different bioactive ingredients including intracellular polysaccharides, adenosine, total polyphenols and triterpenoids in mycelia and extracellular polysaccharides in broth can be obtained from one simple medium for submerged cultivation of X. nigripes.

  19. Sorghum bagasse as substrate for cellulase production by submerged and solid-state cultures of Trichoderma

    Directory of Open Access Journals (Sweden)

    Teodor Vintilă

    2014-05-01

    Full Text Available Sweet sorghum bagasse was used as cellulosic substrate in submerged and solid-state cultures of Trichoderma for cellulase production. Submerged liquid cultures (SLC were obtained by inoculation of Mandels media containing 1% cellulose with spores suspension of Trichoderma. Solid-state cultures (SSC were carried out in Erlenmayer flasks, where the substrate was distributed 1 cm layers. Comparing the yields of cellulases produced by Trichoderma strains in the systems applied in this study, using as substrate sorghum bagasse, we found the solid-state cultures as the system to produce the highest cellulase yields. The local strain of T. viride CMIT3.5. express high productivity in SSC system in laboratory conditions. The cellulolytic enzymes have maximum activity at 50oC, pH 4,8. The results recommend solid-state cultures of Trichoderma on sorghum bagasse as systems for producing cellulolytic products with higher activity than submerged cultures of Trichoderma on the same substrate.

  20. Cow dung is an ideal fermentation medium for amylase production in solid-state fermentation by Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Ponnuswamy Vijayaraghavan

    2015-12-01

    Full Text Available Amylase production by Bacillus cereus IND4 was investigated by solid state fermentation (SSF using cow dung substrate. The SSF conditions were optimized by using one-variable-at-a-time approach and two level full factorial design. Two level full factorial design demonstrated that moisture, pH, fructose, yeast extract and ammonium sulphate have significantly influenced enzyme production (p < 0.05. A central composite design was employed to investigate the optimum concentration of these variables affecting amylase production. Maximal amylase production of 464 units/ml of enzyme was observed in the presence of 100% moisture, 0.1% fructose and 0.01% ammonium sulphate. The enzyme production increased three fold compared to the original medium. The optimum pH and temperature for the activity of amylase were found to be 8.0 and 50 °C, respectively. This enzyme was highly stable at wide pH range (7.0–9.0 and showed 32% enzyme activity after initial denaturation at 50 °C for 1 h. This is the first detailed report on the production of amylase by microorganisms using cow dung as the low cost medium.

  1. Chemistry by nanocatalysis: First example of a solid-supported RAPTA complex for organic reactions in aqueous medium

    KAUST Repository

    García-Garrido, Sergio E.

    2010-11-18

    A ruthenium-arene-PTA (RAPTA) complex has been supported for the first time on an inorganic solid, that is, silica-coated ferrite nanoparticles. The resulting magnetic material proved to be a general, very efficient and easily reusable catalyst for three synthetically useful organic transformations; selective nitrile hydration, redox isomerization of allylic alcohols, and heteroannulation of (Z)-enynols. The use of low metal concentration, environmentally friendly water as a reaction medium, with no use at all of organic solvent during or after the reactions, and microwaves as an alternative energy source renders the synthetic processes reported herein "truly" green and sustainable. RAPTA\\'s delight: A nano-RAPTA complex supported on silica-coated ferrite nanoparticles proved to be a general, very efficient and easily reusable catalyst for three synthetically useful organic transformations; selective nitrile hydration, redox isomerization of allylic alcohols, and heteroannulation of (Z)-enynols. The use of low metal concentrations, water as a reaction medium, and microwaves as an energy source renders these processes green and sustainable. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Optimization of Culture Medium for the Growth of Candida sp. and Blastobotrys sp. as Starter Culture in Fermentation of Cocoa Beans (Theobroma cacao) Using Response Surface Methodology (RSM).

    Science.gov (United States)

    Mahazar, N H; Zakuan, Z; Norhayati, H; MeorHussin, A S; Rukayadi, Y

    2017-01-01

    Inoculation of starter culture in cocoa bean fermentation produces consistent, predictable and high quality of fermented cocoa beans. It is important to produce healthy inoculum in cocoa bean fermentation for better fermented products. Inoculum could minimize the length of the lag phase in fermentation. The purpose of this study was to optimize the component of culture medium for the maximum cultivation of Candida sp. and Blastobotrys sp. Molasses and yeast extract were chosen as medium composition and Response Surface Methodology (RSM) was then employed to optimize the molasses and yeast extract. Maximum growth of Candida sp. (7.63 log CFU mL-1) and Blastobotrys sp. (8.30 log CFU mL-1) were obtained from the fermentation. Optimum culture media for the growth of Candida sp., consist of 10% (w/v) molasses and 2% (w/v) yeast extract, while for Blastobotrys sp., were 1.94% (w/v) molasses and 2% (w/v) yeast extract. This study shows that culture medium consists of molasses and yeast extract were able to produce maximum growth of Candida sp. and Blastobotrys sp., as a starter culture for cocoa bean fermentation.

  3. Influence of culture medium supplementation of tobacco NT1 cell suspension cultures on the N-glycosylation of human secreted alkaline phosphatase.

    Science.gov (United States)

    Becerra-Arteaga, Alejandro; Shuler, Michael L

    2007-08-15

    We report for the first time that culture conditions, specifically culture medium supplementation with nucleotide-sugar precursors, can alter significantly the N-linked glycosylation of a recombinant protein in plant cell culture. Human secreted alkaline phosphatase produced in tobacco NT1 cell suspension cultures was used as a model system. Plant cell cultures were supplemented with ammonia (30 mM), galactose (1 mM) and glucosamine (10 mM) to improve the extent of N-linked glycosylation. The highest levels of cell density and active extracellular SEAP in supplemented cultures were on average 260 g/L and 0.21 U/mL, respectively, compared to 340 g/L and 0.4 U/mL in unsupplemented cultures. The glycosylation profile of SEAP produced in supplemented cultures was determined via electrospray ionization mass spectrometry with precursor ion scanning and compared to that of SEAP produced in unsupplemented cultures. In supplemented and unsupplemented cultures, two biantennary complex-type structures terminated with one or two N-acetylglucosamines and one paucimannosidic glycan structure comprised about 85% of the SEAP glycan pool. These three structures contained plant-specific xylose and fucose residues and their relative abundances were affected by each supplement. High mannose structures (6-9 mannose residues) accounted for the remaining 15% glycans in all cases. The highest proportion (approximately 66%) of a single complex-type biantennary glycan structure terminated in both antennae by N- acetylglucosamine was obtained with glucosamine supplementation versus only 6% in unsupplemented medium. This structure is amenable for in vitro modification to yield a more human-like glycan and could serve as a route to plant cell culture produced therapeutic glycoproteins. (c) 2007 Wiley Periodicals, Inc.

  4. Saccharification of biomass using whole solid-state fermentation medium to avoid additional separation steps.

    Science.gov (United States)

    Pirota, Rosangela D P B; Baleeiro, Flávio C F; Farinas, Cristiane S

    2013-01-01

    The enzymatic hydrolysis of steam-exploded sugarcane bagasse (SESB) was investigated using enzymatic extracts (EE) and whole fermentation media (WM), produced in-house, from Aspergillus niger 3T5B8 and Trichoderma reesei Rut-C30 cultivated on wheat bran under solid-state fermentation (SSF). A detailed and quantitative comparison of the different hydrolysis conditions tested was carried out using the Chrastil approach for modeling enzymatic reactions by fitting the experimental data of total reducing sugar (TRS) released according to hydrolysis time. Conversion of SESB using A. niger enzymatic complex were up to 3.2-fold higher (in terms of TRS) than T. reesei at similar enzyme loadings, which could be correlated to the higher β-glucosidase levels (up to 35-fold higher) of A. niger enzymatic complex. Conversion yields after 72 h exceeded 40% in terms of TRS when the WM was supplemented with a low dosage of a commercial enzyme preparation. When the combination of WM (from either T. reesei or A. niger) and commercial cellulase was used, the dosage of the commercial enzyme could be reduced by half, while still providing a hydrolysis that was up to 36% more efficient. Furthermore, SESB hydrolysis using either EE or WM resulted in similar yields, indicating that the enzyme extraction/filtration steps could be eliminated from the overall process. This procedure is highly advantageous in terms of reduced enzyme and process costs, and also avoids the generation of unnecessary effluent streams. Thus, the enzymatic conversion of SESB using the WM from SSF is cost-effective and compatible with the biorefinery concept. © 2013 American Institute of Chemical Engineers.

  5. Use of secondary sewage water as a culture medium for Chaetoceros gracilis and Thalassiosira Sp (Chrysophyceae in laboratory conditions

    Directory of Open Access Journals (Sweden)

    Rauquírio André Albuquerque Marinho da Costa

    1999-01-01

    Full Text Available Experiments were carried out in order to test the efficiency of additions of secondary sewage as a culture medium for Chaetoceros gracilis and Thalassiosira sp (Chrysophyceae under laboratory conditions. These algae were cultivated in sea water with concentrations of 10%, 20%, 30% and 40% of wastewater. The results were compared with those obtained by the nutritive medium f2 of Guillard (1975. The best results in terms of cellular densities were observed at 40% additions. There were significant differences (significance levels of 5% between the nutritive medium f2 and the 40% additions for both the species. Maximum cellular densities observed for all additions tested were, 4,125.00 x 10³ cells/ml for Chaetoceros gracilis on the ninth day and 834.00 x 10³ cells/ml for Thalassiosira sp on the fifth day. Biomass was higher in the nutritive medium f2 than in the other treatments, reaching average values of 2,363μg/ml for Chaetoceros gracilis. At all experimental units, the best results were registered at 40% addition for Chaetoceros gracilis, where average values of 0.768μg/ml were observed on the fifth day, and at 30% additions for Thalassiosira sp where 0.883μg/ml were observed on the thirteenth day. It was concluded that secondary sewage could be used as a culture medium for the species tested here, after large scale tests.

  6. Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

    Directory of Open Access Journals (Sweden)

    José Soares do Nascimento

    2007-07-01

    Full Text Available The false truffle is one of the main problems in the production of the Agaricus brasiliensis in Brazil and the control of this fungal competitor has been rather difficult due to difficulties in the isolation and cultivation of this pathogen. This experiment was conducted in three stages, the first consisting of the isolation of Diehliomyces microsporus starting from portions of the fruiting body and through the ascospores suspension; second, D. microsporus cultivated in vitro at 15, 20, 25, 30 and 35ºC in six different culture media (CSDA, OCDA, PCDA, ODA, PDA, CDA; third, D. microsporus was inoculated on sterilized compost for formation of the fruiting body. The colony formation from tissue of D. microsporus starting from portions of fruiting body was more efficient than germination of the ascospores. Compost medium (CDA allowed a larger diameter of the D. microsporus colony, followed by the medium made up of compost and potato mixture, favoring a denser composition. The largest mycelial growth speed of D. microsporus occurred when the culture was incubated at 28 and 30ºC. Incubation temperatures lower than 15ºC or above 35ºC inhibited the mycelial growth of D. microsporus completely. The fruiting bodies were obtained easily in sterilized compost and later inoculated along with mycelial competitor.A falsa trufa está sendo um dos principais problemas na produção do Agaricus brasiliensis cultivado no Brasil e o controle deste fungo competidor tem sido difícil, devido às dificuldades encontradas no isolamento e cultivo do patógeno. Este experimento foi conduzido em três etapas, sendo a primeira constituída pelo isolamento de Diehliomyces microsporus a partir de porções do ascostroma e através da suspensão de ascósporos; a segunda, o cultivo in vitro de D. microsporus nas temperaturas de 15, 20, 25, 30 e 35ºC e em seis meios de cultura (CTDA, ACDA, BCDA, ADA, BDA e CDA e a terceira pela inoculação de D. microsporus no composto

  7. A microPIXE investigation of the interaction of cells of Schizosaccharomyces pombe with the culture medium

    Energy Technology Data Exchange (ETDEWEB)

    Rombouts, P.M.M. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Gomez-Morilla, I. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Grime, G.W. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Webb, R.P. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Cuenca, L. [Fluids and Systems Research Centre, School of Engineering, University of Surrey, Guildford GU2 7XH (United Kingdom); Rodriguez, R. [Fluids and Systems Research Centre, School of Engineering, University of Surrey, Guildford GU2 7XH (United Kingdom); Browton, M. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Wardell, N. [School of Biomedical and Molecular Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom); Underwood, B. [Fluids and Systems Research Centre, School of Engineering, University of Surrey, Guildford GU2 7XH (United Kingdom); Kirkby, N.F. [Fluids and Systems Research Centre, School of Engineering, University of Surrey, Guildford GU2 7XH (United Kingdom); Kirkby, K.J. [Ion Beam Centre, Advanced Technology Institute, School of Electronics and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom)]. E-mail: k.kirkby@surrey.ac.uk

    2007-07-15

    Schizosaccharomyces pombe (S. pombe) is a eucaryotic cell type similar to mammalian cells but much more simple. As it also executes its cell cycle rapidly it is very useful for investigating basic processes in cells. In this paper we report a feasibility study of the applicability of microPIXE to investigate the interaction between S. pombe cells and the surrounding culture medium. Cells were cultured in various growth medium prior to preparation for analysis. 1 {mu}l drops of medium and cells were spotted onto polypropylene foils held in contact with a polished copper block previously cooled in liquid nitrogen. The samples were dehydrated by freeze-drying. Micro PIXE analysis was carried out with the IBC microbeam facility using a beam of 2.5 MeV protons focused to 1-2 {mu}m diameter. Initially no elemental contrast was observed between the cells and the medium, but by modifying the dilution of the cell suspension, the cells could be distinguished from the surrounding medium through an increased concentration of P and reduced concentration of Cl. The distribution of Na in the medium around the cells showed evidence of the action of the Na pump. Sporulation appears to be induced in the cells by adding Cu to the growth medium and the uptake of Cu by the cells could be clearly observed. This study shows that it is possible to analyse the mass transport of elements in and out of cells In the future this will enable concentration gradients to be analysed and allow the rate of production or consumption of individual cells to be calculated. By observing these patterns for individual cells (not populations) at various known points in the cell cycle, fundamental data can be derived.

  8. Using CR1aa versus KSOM as the culture medium for in vitro embryo production of cattle

    Directory of Open Access Journals (Sweden)

    Endang Triwulaninngsih

    2002-03-01

    Full Text Available This research has been conducted at the laboratory of in vitro fertilization in the Department of Animal Science University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. Oocytes were matured in TCM- 199 medium (in 5% CO2 incubator and at 390C enriched with follicle stimulating hormone (FSH 10 μl/ml, oestradiol 17 β 1μl/ml and 10% Fetal Calf Serum (FCS. The oocytes were fertilized in vitro with motile sperm and incubation between sperm and oocytes in fertilization medium Tyroide Albumin Lactate Pyruvate (TALP for 20 hours. All zygotes were cultured in CR1aa (n=1549 medium versus modification of protein-free pottasium simplex optimized medium (KSOM (n=675 up to blastocyst stage and were fed FCS 5 μl/50 μl medium on day 6, as treatment A and B respectively. Data were analyzed by completely randomized design with SAS program. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 91.4% vs 75.6 %; 75.6% vs 58.9%; 61.5% vs 38.5%; 31.2% vs 5.1%, 8.6% vs 24.4%, 15.7% vs 8% which were significantly different (P<0.01 for treatment CR1aa and KSOM respectively. Based on this study, CR1aa medium is better culture medium than KSOM for efficient in vitro production (IVP of bovine embryos.

  9. A microPIXE investigation of the interaction of cells of Schizosaccharomyces pombe with the culture medium

    International Nuclear Information System (INIS)

    Rombouts, P.M.M.; Gomez-Morilla, I.; Grime, G.W.; Webb, R.P.; Cuenca, L.; Rodriguez, R.; Browton, M.; Wardell, N.; Underwood, B.; Kirkby, N.F.; Kirkby, K.J.

    2007-01-01

    Schizosaccharomyces pombe (S. pombe) is a eucaryotic cell type similar to mammalian cells but much more simple. As it also executes its cell cycle rapidly it is very useful for investigating basic processes in cells. In this paper we report a feasibility study of the applicability of microPIXE to investigate the interaction between S. pombe cells and the surrounding culture medium. Cells were cultured in various growth medium prior to preparation for analysis. 1 μl drops of medium and cells were spotted onto polypropylene foils held in contact with a polished copper block previously cooled in liquid nitrogen. The samples were dehydrated by freeze-drying. Micro PIXE analysis was carried out with the IBC microbeam facility using a beam of 2.5 MeV protons focused to 1-2 μm diameter. Initially no elemental contrast was observed between the cells and the medium, but by modifying the dilution of the cell suspension, the cells could be distinguished from the surrounding medium through an increased concentration of P and reduced concentration of Cl. The distribution of Na in the medium around the cells showed evidence of the action of the Na pump. Sporulation appears to be induced in the cells by adding Cu to the growth medium and the uptake of Cu by the cells could be clearly observed. This study shows that it is possible to analyse the mass transport of elements in and out of cells In the future this will enable concentration gradients to be analysed and allow the rate of production or consumption of individual cells to be calculated. By observing these patterns for individual cells (not populations) at various known points in the cell cycle, fundamental data can be derived

  10. A novel liquid medium for the efficient growth of the salmonid pathogen Piscirickettsia salmonis and optimization of culture conditions.

    Directory of Open Access Journals (Sweden)

    Mirtha Henríquez

    Full Text Available Piscirickettsia salmonis is the bacterium that causes Piscirickettsiosis, a systemic disease of salmonid fish responsible for significant economic losses within the aquaculture industry worldwide. The growth of the bacterium for vaccine formulation has been traditionally accomplished by infecting eukaryotic cell lines, a process that involves high production costs and is time-consuming. Recent research has demonstrated that it is possible to culture pure P. salmonis in a blood containing (cell-free medium. In the present work we demonstrate the growth of P. salmonis in a liquid medium free from blood and serum components, thus establishing a novel and simplified bacteriological medium. Additionally, the new media reported provides improved growth conditions for P. salmonis, where biomass concentrations of approximately 800 mg cell dry weight L(-1 were obtained, about eight times higher than those reported for the blood containing medium. A 2- level full factorial design was employed to evaluate the significance of the main medium components on cell growth and an optimal temperature range of 23-27°C was determined for the microorganism to grow in the novel liquid media. Therefore, these results represent a breakthrough regarding P. salmonis research in order to optimize pure P. salmonis growth in liquid blood and serum free medium.

  11. A randomized clinical trial to evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium for in vitro fertilization

    DEFF Research Database (Denmark)

    Ziebe, Søren; Loft, Anne; Povlsen, Betina B

    2013-01-01

    To evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium on ongoing implantation rate (OIR).......To evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium on ongoing implantation rate (OIR)....

  12. Development of a mixed culture chain elongation process based on municipal solid waste and ethanol

    NARCIS (Netherlands)

    Grootscholten, T.I.M.

    2013-01-01

    Keywords: mixed culture fermentation; Carboxylates; Caproate; Heptanoate; ethanol; OFMSW

    To reduce dependence on oil, alternative fuel and chemical production processes are investigates. In this thesis, we investigated the production of medium chain fatty acids (MCFAs) using an anaerobic

  13. Influence of embryo culture medium (G5 and HTF) on pregnancy and perinatal outcome after IVF: a multicenter RCT.

    Science.gov (United States)

    Kleijkers, Sander H M; Mantikou, Eleni; Slappendel, Els; Consten, Dimitri; van Echten-Arends, Jannie; Wetzels, Alex M; van Wely, Madelon; Smits, Luc J M; van Montfoort, Aafke P A; Repping, Sjoerd; Dumoulin, John C M; Mastenbroek, Sebastiaan

    2016-10-01

    Does embryo culture medium influence pregnancy and perinatal outcome in IVF? Embryo culture media used in IVF affect treatment efficacy and the birthweight of newborns. A wide variety of culture media for human preimplantation embryos in IVF/ICSI treatments currently exists. It is unknown which medium is best in terms of clinical outcomes. Furthermore, it has been suggested that the culture medium used for the in vitro culture of embryos affects birthweight, but this has never been demonstrated by large randomized trials. We conducted a multicenter, double-blind RCT comparing the use of HTF and G5 embryo culture media in IVF. Between July 2010 and May 2012, 836 couples (419 in the HTF group and 417 in the G5 group) were included. The allocated medium (1:1 allocation) was used in all treatment cycles a couple received within 1 year after randomization, including possible transfers with frozen-thawed embryos. The primary outcome was live birth rate. Couples that were scheduled for an IVF or an ICSI treatment at one of the six participating centers in the Netherlands or their affiliated clinics. The live birth rate was higher, albeit nonsignificantly, in couples assigned to G5 than in couples assigned to HTF (44.1% (184/417) versus 37.9% (159/419); RR: 1.2; 95% confidence interval (CI): 0.99-1.37; P = 0.08). Number of utilizable embryos per cycle (2.8 ± 2.3 versus 2.3 ± 1.8; P culture media on perinatal outcome remains to be determined. Embryo culture media used in IVF affect not only treatment efficacy but also perinatal outcome. This suggests that the millions of human embryos that are cultured in vitro each year are sensitive to their environment. These findings should lead to increased awareness, mechanistic studies and legislative adaptations to protect IVF offspring during the first few days of their existence. This project was partly funded by The NutsOhra foundation (Grant 1203-061) and March of Dimes (Grant 6-FY13-153). The authors declare no conflict of

  14. Change in enzyme production by gradually drying culture substrate during solid-state fermentation.

    Science.gov (United States)

    Ito, Kazunari; Gomi, Katsuya; Kariyama, Masahiro; Miyake, Tsuyoshi

    2015-06-01

    The influence of drying the culture substrate during solid-state fermentation on enzyme production was investigated using a non-airflow box. The drying caused a significant increase in enzyme production, while the mycelium content decreased slightly. This suggests that changes in the water content in the substrate during culture affect enzyme production in fungi. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  15. Radiation-induced organogenesis: effects of irradiated medium and its components on tobacco tissue culture

    International Nuclear Information System (INIS)

    Degani, N.

    1975-01-01

    Gamma irradiated medium induces the formation of buds in non-irradiated dark growth tobacco callus (Nicotiana tabacum Var. Wisconsin No.38). Experiments were conducted to determine the component(s) of the medium that is effective in this radiation-induced organogenesis. Fraction of medium were irradiated singly and in combination, then combined with non-irradiated fractions to form the complete growth medium. The results showed that irradiated indoleacetic acid (IAA) was not the effective component in the induction of organogensis. Omission of IAA from the medium resulted in the formation of buds, as expected. Irradiated myo-inositol induced organogenesis more consistently than the other irradiated components. The age of the inoculum tissue and its passage number from the tobacco stem affected the potency of the tobacco callus to organise. (author)

  16. Impact of culture medium on maturation of bone marrow-derived murine dendritic cells via the aryl hydrocarbon receptor.

    Science.gov (United States)

    Ilchmann, Anne; Krause, Maren; Heilmann, Monika; Burgdorf, Sven; Vieths, Stefan; Toda, Masako

    2012-05-01

    The aryl hydrocarbon receptor (AhR) plays a role in modulating dendritic cell (DC) immunity. Iscove's modified Dulbecco's medium (IMDM) contains higher amounts of AhR ligands than RPMI1640 medium. Here, we examined the influence of AhR ligand-containing medium on the maturation and T-cell stimulatory capacity of bone marrow-derived murine dendritic cells (BMDCs). BMDCs generated in IMDM (BMDCs/IMDM) expressed higher levels of co-stimulatory and MHC class II molecules, and lower levels of pattern-recognition receptors, especially toll-like receptor (TLR) 2, TLR4, and scavenger receptor class A (SR-A), compared to BMDCs generated in RPMI1640 medium (BMDCs/RPMI). Cytokine responses against ligands of TLRs and antigen uptake mediated by SR-A were remarkably reduced in BMDCs/IMDM, whereas the T-cell stimulatory capacity of the cells was enhanced, compared to BMDCs/RPMI. The enhanced maturation of BMDCs/IMDM was attenuated in the presence of an AhR antagonist, indicating involvement of AhR in the maturation. Interestingly, BMDCs/IMDM induced Th2 and Th17 differentiation at low and high concentrations of antigen respectively, when co-cultured with CD4(+) T-cells from antigen-specific T-cell receptor transgenic mice. In contrast, BMDCs/RPMI induced Th1 differentiation predominantly in the co-culture. Taken together, optimal selection of medium seems necessary when studying BMDCs, depending on the target receptors on the cell surface of DCs and type of helper T-cells for the co-culture. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture.

    Science.gov (United States)

    Fan, Yuzhou; Kildegaard, Helene Faustrup; Andersen, Mikael Rørdam

    2017-01-01

    Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. In this chapter, we will focus on how to carry out experiments for N-glycosylation modulation through medium and feed optimization. The workflow and typical methods involved in the experiment process will be presented.

  18. Influence of the Addition of Riboflavin in Culture Medium on Delivering Biomass Using Yeast Strains of Saccharomyces Carlsbengensis

    Directory of Open Access Journals (Sweden)

    Cornelia Nicoară

    2010-05-01

    Full Text Available Yeasts requirements for growth factors should be considered both in terms of ability to summarize the simpleaverage and the dependence on external supplies. Vitamins are components of coenzymes or enzymes prostheticgroups and thus they are growth factors for yeast. The study concerns about the influence of the addition ofriboflavin in culture medium in different quantities, the accumulation of yeast biomass under the action of yeaststrains of beer. The process of cultivation has been made for 24 hours at a temperature of 220C. The addition ofriboflavin in culture medium of yeast biomass increased in each strain of yeast compared with the witness - thesample without added riboflavin. Biomass obtained by follow this procedure could be used to create new foodproducts with high ration nutritional value.

  19. Successful non-surgical deep uterine transfer of porcine morulae after 24 hour culture in a chemically defined medium.

    Directory of Open Access Journals (Sweden)

    Emilio A Martinez

    Full Text Available Excellent fertility and prolificacy have been reported after non-surgical deep uterine transfers of fresh in vivo-derived porcine embryos. Unfortunately, when this technology is used with vitrified embryos, the reproductive performance of recipients is low. For this reason and because the embryos must be stored until they are transferred to the recipient farms, we evaluated the potential application of non-surgical deep uterine transfers with in vivo-derived morulae cultured for 24 h in liquid stage. In Experiment 1, two temperatures (25 °C and 37 °C and two media (one fully defined and one semi-defined were assessed. Morulae cultured in culture medium supplemented with bovine serum albumin and fetal calf serum at 38.5 °C in 5% CO2 in air were used as controls. Irrespective of medium, the embryo viability after 24 h of culture was negatively affected (P<0.05 at 25 °C but not at 37 °C compared with the controls. Embryo development was delayed in all experimental groups compared with the control group (P<0.001. Most of the embryos (95.7% cultured at 37 °C achieved the full or expanded blastocyst stage, and unlike the controls, none of them hatched at the end of culture. In Experiment 2, 785 morulae were cultured in the defined medium at 37 °C for 24 h, and the resulting blastocysts were transferred to the recipients (n = 24. Uncultured embryos collected at the blastocyst stage (n = 750 were directly transferred to the recipients and used as controls (n = 25. No differences in farrowing rates (91.7% and 92.0% or litter sizes (9.0 ± 0.6 and 9.4 ± 0.8 were observed between the groups. This study demonstrated, for the first time, that high reproductive performance can be achieved after non-surgical deep uterine transfers with short-term cultured morulae in a defined medium, which opens new possibilities for the sanitary, safe national and international trade of porcine embryos and the commercial use of embryo transfer in pigs.

  20. Engineer medium and feed for modulating N-glycosylation of recombinant protein production in CHO cell culture

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Kildegaard, Helene Faustrup; Andersen, Mikael Rørdam

    2017-01-01

    Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. Control and optimization of protein N......-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell...

  1. Bicarbonate Plays a Critical Role in the Generation of Cytotoxicity during SIN-1 Decomposition in Culture Medium

    Directory of Open Access Journals (Sweden)

    Kyo Shirai

    2012-01-01

    Full Text Available 3-Morpholinosydnonimine (SIN-1 is used as a donor of peroxynitrite (ONOO− in various studies. We demonstrated, however, that, the cell-culture medium remains cytotoxic to PC12 cells even after almost complete SIN-1 decomposition, suggesting that reaction product(s in the medium, rather than ONOO−, exert cytotoxic effects. Here, we clarified that significant cytotoxicity persists after SIN-1 decomposes in bicarbonate, a component of the culture medium, but not in NaOH. Cytotoxic SIN-1-decomposed bicarbonate, which lacks both oxidizing and nitrosating activities, degrades to innocuous state over time. The extent of SIN-1 cytotoxicity, irrespective of its fresh or decomposed state, appears to depend on the total number of initial SIN-1 molecules per cell, rather than its concentration, and involves oxidative/nitrosative stress-related cell damage. These results suggest that, despite its low abundance, the bicarbonate-dependent cytotoxic substance that accumulates in the medium during SIN-1 breakdown is the cytotoxic entity of SIN-1.

  2. Application of Statistical Design to the Optimization of Culture Medium for Prodigiosin Production by Serratia marcescens SWML08

    OpenAIRE

    Venil, C. K.; Lakshmanaperumalsamy, P.

    2009-01-01

    Combination of Plackett – Burman design (PBD) and Box – Behnken design (BBD) were applied for optimization of different factors for prodigiosin production by Serratia marcescens SWML08. Among 11 factors, incubation temperature, and supplement of (NH4)2PO4 and trace salts into the culture medium were selected due to significant positive effect on prodigiosin yield. Box - Behnken design, a response surface methodology, was used for further optimization of these selected factors for better pro...

  3. Determination of free cisplatin in medium by differential pulse polarography after ultrasound and cisplatin treatment of a cancer cell culture

    International Nuclear Information System (INIS)

    Bernard, Vladan; Skorpikova, Jirina; Mornstein, Vojtech; Fojt, Lukas

    2011-01-01

    The in vitro study was carried out for detection of the cisplatin in free form and in culture medium, depending on various conditions of sonodynamic human ovarian cancer cells A2780 treatment by differential pulse polarography (DPP). For sonodynamic treatment, we used cisplatin alone and combined cisplatin/ultrasound treatments. The ultrasound exposure intensity of 1.0 and 2.0 Wcm 2 in far field for incubation periods 1, 24 and 48 h was used. The parameters of DPP measurements were - 1 s drop time, 5 mV.s -1 voltage scan rate, 50 mV modulation amplitude and negative scanning direction; platinum wire served as counter electrode and Ag|AgCl|3 M KCI as reference electrode. The results showed the dependence of free platinum quantities in culture medium on incubation time and treatment protocol. We found difference in concentration of free cisplatin between conventional application of cisplatin and sonodynamic treatment. The sonodynamic combined treatment of cisplatin and ultrasound field showed a higher cisplatin content in the culture medium than cisplatin treatment alone; a difference of 20% was observed for incubation time 48 h. The results also showed the influence of a time sequence of ultrasound and cytostatics in the sonodynamic treatment. The highest amount of free cisplatin in the solution was found for primary application of cisplatin and the subsequent ultrasound exposure. The quantity of free cisplatin increased with time, namely for time intervals 1-24 h. There was no difference between the DPP signal of cisplatin in reaction mixture containing cells in small quantities and micro-filtered mixture without cells. Thus, the DPP method is suitable for the detection and quantification of free cisplatin in the culture medium of cell suspension. Ultrasound field can be important factor during cytostatic therapy. (author)

  4. Production of Trametes pubescens Laccase under Submerged and Semi-Solid Culture Conditions on Agro-Industrial Wastes

    Science.gov (United States)

    Rodriguez, Alexander; Osma, Johann F.; Alméciga-Díaz, Carlos J.; Sánchez, Oscar F.

    2013-01-01

    Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametes pubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69±0.28 U mg-1 of protein for the crude extract, and 0.08±0.001 and 2.86±0.05 U mg-1 of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct. PMID:24019936

  5. Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes.

    Science.gov (United States)

    Gonzalez, Juan C; Medina, Sandra C; Rodriguez, Alexander; Osma, Johann F; Alméciga-Díaz, Carlos J; Sánchez, Oscar F

    2013-01-01

    Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametespubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69 ± 0.28 U mg(-1) of protein for the crude extract, and 0.08 ± 0.001 and 2.86 ± 0.05 U mg(-1) of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

  6. Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes.

    Directory of Open Access Journals (Sweden)

    Juan C Gonzalez

    Full Text Available Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametespubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM, and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1 and 60 kDa (Lac2. Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69 ± 0.28 U mg(-1 of protein for the crude extract, and 0.08 ± 0.001 and 2.86 ± 0.05 U mg(-1 of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

  7. Accurate and noninvasive embryos screening during in vitro fertilization (IVF) assisted by Raman analysis of embryos culture medium

    International Nuclear Information System (INIS)

    Shen, A G; Peng, J; Su, L; Wang, X H; Hu, J M; Zhao, Q H; Yang, J

    2012-01-01

    In combination with morphological evaluation tests, we employ Raman spectroscopy to select higher potential reproductive embryos during in vitro fertilization (IVF) based on chemical composition of embryos culture medium. In this study, 57 Raman spectra are acquired from both higher and lower quality embryos culture medium (ECM) from 10 patients which have been preliminarily confirmed by clinical assay. Data are fit by using a linear combination model of least squares method in which 12 basis spectra represent the chemical features of ECM. The final fitting coefficients provide insight into the chemical compositions of culture medium samples and are subsequently used as criterion to evaluate the quality of embryos. The relative fitting coefficients ratios of sodium pyruvate/albumin and phenylalanine/albumin seem act as key roles in the embryo screening, attaining 85.7% accuracy in comparison with clinical pregnancy. The good results demonstrate that Raman spectroscopy therefore is an important candidate for an accurate and noninvasive screening of higher quality embryos, which potentially decrease the time-consuming clinical trials during IVF

  8. Solutions to academic failure: The cognitive and cultural realities ofEnglish as the medium of instruction among black learners

    Directory of Open Access Journals (Sweden)

    R. Gamaroff

    2013-02-01

    Full Text Available In South Africa, black learners who are speakers of Bantu languages have to use a second language, namely English, as the medium of instruction from Std 3 onwards. The differences between English language-culture and Bantu languages-culture(s have generated a host of problems (and pseudo-problems?, where the main problem is academic failure. Three solutions to academic failure are discussed in the light of cultural and cognitive factors in multicultural education: 1. The use of the mother tongue as the exclusive medium of instruction 2. Critical Language Study (CLS and People's English 3. The separation of high ability learners from limited ability learners in the teaching situation. It is emphasised that culture is closely connected to a symbolic system, and thus an understanding of cognitive processes in academic learning requires an understanding of culture, and vice versa. Ultimately, of primary importance in academic study are the cognitive underpinnings of Cognitive Academic Language Proficiency (CALP developed in the first language. In Suid-Afrika word swart leerders wie se moedertaal een van die Afrika tale is, tans vanaf st. 3 in 'n tweede taal, naamlik Engels, onderrig. As gevolg van die verskille tussen die Engelse taalkultuur en die taalkulture van die A.frika tale het daar 'n groot aantal probleme (en pseudoprobleme? ontstaan, waarvan akademiese mislukking die belangrikste is. Drie oplossings vir hierdie akademiese mislukking word bespreek aan die hand van kulturele en kognitiewe faktore in multikulturele onderwys: 1. Die gebruik van die moedertaal as eksklusiewe medium van onderrig 2. "Critical Language Study" (CLS en "People's English" 3. Die afsonderlike hantering van hoogsbegaafde en minder begaafde leerlinge. Dit moet beklemtoon word dat kultuur nouverwant is aan 'n simbolesisteem. Gevolglik is 'n be grip van die kognitiewe prosesse betrokke by akademiese leer 'n voorvereiste vir 'n be grip van kultuur, en omgekeerd. Vera

  9. The Influence of the Intangible Cultural Heritage About propagation medium in the Big data Era

    Directory of Open Access Journals (Sweden)

    Jin Zhuo

    2016-01-01

    Full Text Available Film and television works have been attracting the attention of the modern people with their own characteristics. Image, vivid, intuitive, popular, the media uses its own advantage to spread out the information quickly, the transmission efficiency and the promotion is unmatched by other any medias, it also plays an important role for the dissemination of intangible cultural heritage. With our country pay more attention on protecting the intangible cultural heritage, digital media can use its own advantages to protect, promote, promote the intangible cultural heritage, to visual intangible cultural heritage, and present it to the public, in order to let the public understand what is intangible cultural heritage, take the initiative to protect the intangible cultural heritage. This paper discusses the influence of digital media on protecting intangible cultural heritage, with the power of digital media, we expect to protect it better, leave a valuable legacy for future generations.

  10. Effect of Somatic Cell Types and Culture Medium on in vitro Maturation, Fertilization and Early Development Capability of Buffalo Oocytes

    Directory of Open Access Journals (Sweden)

    H. Jamil*, H. A. Samad, N. Rehman, Z. I. Qureshi and L. A. Lodhi

    2011-04-01

    Full Text Available This study was designed to evaluate the efficacy of different somatic cell types and media in supporting in vitro maturation (IVM, in vitro fertilization (IVF and early embryonic development competence of buffalo follicular oocytes. Cumulus oocyte complexes were collected for maturation from follicles (>6mm of buffalo ovaries collected at the local abattoir. Oocytes were co-cultured in tissue culture medium (TCM-199 with either granulosa cells, cumulus cells, or buffalo oviductal epithelial cells (BOEC @ 3x106 cells/ml or in TCM-199 without helper cells (control at 39°C and 5%CO2 in humidified air. Fresh semen was prepared in modified Ca++ free Tyrode medium. Fertilization was carried out in four types of media: i Tyrode lactate albumin pyruvate (TALP, ii TALP+BOEC, iii modified Ca++ free Tyrode and iv modified Ca++ free Tyrode+BOEC. Fertilized oocytes were cultured for early embryonic development in TCM-199 with and without BOEC. Higher maturation rates were observed in the granulosa (84.24% and cumulus cells (83.44% than BOEC co culture system (73.37%. Highest fertilization rate was obtained in modified Ca++ free Tyrode with BOEC co culture (70.42%, followed by modified Ca++ free Tyrode alone (63.77%, TALP with BOEC (36.92% and TALP alone (10.94%. Development of early embryos (8-cell stage improved in TCM-199 with BOEC co culture than TCM-199 alone. From the results of this study, it can be concluded that addition of somatic cells (granulosa cells, cumulus cells results in higher maturation rates of buffalo follicular oocytes than BOEC co culture system, while fertilization rate improved in modified Ca++ free Tyrode with and without BOEC. Addition of BOEC to TCM-199 improved the developmental capacity of early embryo.

  11. Effects of polyurethane matrices on fungal tannase and gallic acid production under solid state culture

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The influence of the physical structure of polyurethane matrix as a support in a solid state culture in tannase production and gallic acid accumulation by Aspergillus niger Aa-20 was evaluated. Three different polyurethane matrices were used as the support: continuous, semi-discontinuous and discontinuous. The highest tannase production at 2479.59 U/L during the first 12 h of culture was obtained using the discontinuous matrix. The gallic acid was accumulated at 7.64 g/L at the discontinuous matrix. The results show that the discontinuous matrix of polyurethane is better for tannase production and gallic acid accumulation in a solid state culture bioprocess than the continuous and semi-discontinuous matrices.

  12. Determination better culture medium in the establishment phase for the in vitro propagation of banana (Musa paradisiaca L

    Directory of Open Access Journals (Sweden)

    Ancasi-Espejo Ruth Gabriela

    2016-08-01

    Full Text Available This research was conducted at the Laboratory of Plant Biotechnology of the Department of Biological and Natural Sciences of the Amazonian University of Pando, in 2014. The aim of the study was to determine better culture medium in the establishment phase for propagation in vitro banana (Musa paradisiaca L., 20 were selected and characterized mother plants NTRCA (New Technology Research Center Amazonia. A completely random design (CRD with three different culture media was used. The culture media were M1 Murashige and Skoog (MS was supplemented with ascorbic acid 100 mg/L and L-cysteine 2 ml /L, M2 Murashige and Skoog (MS was supplemented charcoal 2 g/L, M3 Murashige and Skoog (MS supplement-ed with ascorbic acid 100 mg/L and cítrico100 mg/L acid. The variables evaluated were: The survival of the former Plantes, where contamination and oxidation was observed. The results showed that in the first phase of establishment, the best answer for the survival of the former Plantes banana (Musa paradisiaca, was with the culture medium 3, where a lower degree of oxidation (0.26 and pollution for all explants was obtained was 28%.

  13. Interactions between Candida albicans and Candida glabrata in biofilms: Influence of the strain type, culture medium and glucose supplementation.

    Science.gov (United States)

    Hosida, Thayse Yumi; Cavazana, Thamires Priscila; Henriques, Mariana; Pessan, Juliano Pelim; Delbem, Alberto Carlos Botazzo; Monteiro, Douglas Roberto

    2018-04-01

    The relationship among Candida species may be influenced by several factors. Thus, this study evaluated the interactions between Candida albicans and Candida glabrata in biofilms, varying the strain type, culture medium and glucose supplementation. Biofilms were formed for 48 hours in Sabouraud dextrose broth (SDB) or RPMI 1640, supplemented with 0%, 1% or 5% glucose. Each strain of C. albicans was combined with two strains of C. glabrata, generating four biofilm associations, which were quantified by colony-forming units (CFUs), total biomass and metabolic activity. Data were analysed by ANOVA and Tukey's HSD test (α = 0.05). For CFUs, all associations were classified as indifferent for biofilms formed in RPMI 1640, while for SDB the interactions were antagonistic for C. albicans and indifferent for C. glabrata. The association of reference strains resulted in a dual-species biofilm with biomass significantly higher than that observed for each single biofilm developed in SDB. The metabolic activity of dual-species biofilms did not significantly differ from that found for single ones, except for co-culture of the reference strains. Glucose supplementation and culture media had a significant influence on all parameters. In conclusion, the strain type, culture medium and glucose supplementation influenced the interactions between C. albicans and C. glabrata. © 2017 Blackwell Verlag GmbH.

  14. IVF culture medium affects post-natal weight in humans during the first 2 years of life.

    Science.gov (United States)

    Kleijkers, Sander H M; van Montfoort, Aafke P A; Smits, Luc J M; Viechtbauer, Wolfgang; Roseboom, Tessa J; Nelissen, Ewka C M; Coonen, Edith; Derhaag, Josien G; Bastings, Lobke; Schreurs, Inge E L; Evers, Johannes L H; Dumoulin, John C M

    2014-04-01

    Is post-natal growth during the first 2 years of life in IVF singletons affected by type of medium used for culturing human embryos during an IVF treatment? The in vitro culture of human embryos in medium from Cook resulted in singletons with a lower weight during the first 2 years of life compared with singletons born after embryo culture in medium from Vitrolife. In a previous study, we reported that type of medium used for culturing human IVF embryos during the first few days after fertilization until fresh embryo transfer significantly affects fetal growth and consequently birthweight of the resulting singletons. From July 2003 to December 2006, a total of 1432 IVF treatment cycles with fresh embryo transfer were randomly allocated to have all embryos cultured in medium from Vitrolife AB (n = 715) or from Cook (n = 717). Two years after delivery, questionnaires were sent to the parents of all children requesting data about weight, height and head circumference around 1, 2, 3, 4, 6, 7.5, 9, 11, 14, 18 and 24 months of age. These measurements were collected as part of the children's health programme at municipal infant welfare centres in the Netherlands by health professionals unaware of this study. Patients requiring donor oocytes or applying for PGD were excluded from the study. From the 294 live born singletons that fulfilled our inclusion criteria, 29 were lost to follow-up. The remaining 265 singletons (Cook group: 117, Vitrolife group: 148) were included in the analysis. Data analysis included linear regression, to compare cross-sectionally weight standard deviation score (SDS), height SDS and head circumference, and the first order Berkey-Reed model for a longitudinal analysis of the growth data. Singletons in the Vitrolife group were heavier during the first 2 years of life compared with singletons in the Cook group. Cross-sectional analyses showed that adjusted weight SDS differed between groups at 1 (0.35 ± 0.14, P = 0.010), 2 (0.39 ± 0.14, P = 0

  15. Optimization of flask culture medium and conditions for hyaluronic acid production by a streptococcus equisimilis mutant nc2168

    Directory of Open Access Journals (Sweden)

    Yong-Hao Chen

    2012-12-01

    Full Text Available A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116by ultraviolet ray combined with60Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA. In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33ºC, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.

  16. THE INFLUENCE OF THE COMPOSITION OF THE CULTURE MEDIUM ON THE DEVELOPMENT OF LEUCONOSTOC LACTIS PRE-FERMENTATION

    Directory of Open Access Journals (Sweden)

    V. V. Kondratenko

    2017-01-01

    Full Text Available The regularity of the influence of the culture medium (substrate on the development of microflora at the stage of preliminary fermentation of the model medium on the basis of white cabbage varieties "Parus" was studied. During the research, strains of lactic acid microorganisms Leuconostoc lactis were used. Step-by-step mathematical processing of the experimental data was carried out. Functional dependencies are obtained that most adequately approximate experimental data for modified (MMC and basic (BMS model media. Analysis of the experimental data showed that, depending on the type (composition of the medium, the same species of microorganisms exhibit different dynamics of titer growth. In connection with this, an algorithm was developed to determine the optimal duration of pre-fermentation – «stop points». As a result of the research, it can be seen that the modification of the model medium with the addition of table salt and ascorbic acid to it promotes the formation of positive dynamics of the comparison indicator. This dynamics has three extremes, but only extremes are of practical significance, which were in the interval of the monotonic decrease of the titer. For successful development of the starting culture of the stage of the main fermentation, one of the conditions is a relatively small amount of the titer of the first culture at the end of the preliminary fermentation step to exclude competition. Thus, the position of the «stop-point» position corresponds to the period after the last peak of the comparison indicator. The investigated regularity of the effect of the preliminary cultivation of gram-positive microorganisms on the activity of lactic acid microorganisms in the process of fermentation is topical, since the whole process and the production of high-quality products fully depend on this approach.

  17. In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum.

    Science.gov (United States)

    Bruserud, Oystein; Tronstad, Karl Johan; Berge, Rolf

    2005-06-01

    Experimental in vitro models including well-characterised cell lines can be used to identify possible new therapeutic targets for the treatment of osteosarcoma. Culture media including inactivated serum is often recommended for in vitro culture of osteosarcoma cells, but the serum component then represents a nonstandardised parameter including a wide range of unidentified mediators. To improve the standardisation we have investigated whether serum-free culture media can be used in experimental in vitro studies of osteosarcoma cell lines. The seven osteosarcoma cell lines Cal72, SJSA-1, Saos-2, SK-ES-1, U2OS, 143.98.2, and KHOS-32IH were cultured in vitro in various serum-free media and media supplemented with 10% heat-inactivated fetal calf serum (FCS). Although proliferation often was relatively low in serum-free media (X-vivo 10, X-vivo 15, X-vivo 20, Stem Span SFEM), some cell lines (Cal72, KHOS-32IH, Saos-2) showed proliferation comparable with the recommended FCS-containing media even when using serum-free conditions. The optimal serum-free medium then varied between cell lines. We also compared 6 different FCS-containing media (including Stem Span with 10% FCS) and the optimal FCS-containing medium varied between cell lines. However, all cell lines proliferated well in Stem Span with FCS, and this medium was regarded as optimal for four of the lines. FCS could not be replaced by fatty acids or low density lipoprotein when testing the Stem Span medium. The release of a wide range of soluble mediators showed only minor differences when using serum-free and FCS-containing media (including Stem Span with and without FCS), and serum-free Stem Span could also be used for in vitro studies of mitogen-stimulated T cell activation in the presence of accessory osteosarcoma cells. The use of Stem Span with 10% FCS allowed the release of a wide range of chemokines by osteosarcoma cell lines (Cal72, SJSA-1), and the chemokine release profile was very similar to the

  18. Comparison of tumour age response to radiation for cells derived from tissue culture or solid tumours

    International Nuclear Information System (INIS)

    Keng, P.C.; Siemann, D.W.; Rochester Univ., NY; Rochester Univ., NY; Wheeler, K.T.

    1984-01-01

    Direct comparison of the cell age response of 9L and KHT tumour cells derived either from tissue culture or solid tumours was achieved. Cells from dissociated KHT and 9L tumours (the latter implanted either subcutaneously or intracerebrally) and cells from tissue culture were separated into homogenous sized populations by centrifugal elutriation. In both tumour models these homogeneous sized populations correspond to populations enriched at different stages of the cell cycle. The survival of these elutriated cell populations was measured after a single dose of Cs-137 gamma rays. For cells isolated from 9L solid tumours, there was little variation in radiosensitivity throughout the cell cycle; however, a very small but significant increase in resistance was found in late G 1 cells. This lack of a large variation in radiosensitivity through the cell cycle for 9L cells from solid tumours also was seen in 9L cells growing in monolayer tissue culture. When similar experiments were performed using the KHT sarcoma tumour model, the results showed that KHT cells in vitro exhibited a fairly conventional increase in radioresistance in both mid G 1 and late S. However, the cell age response of KHT cells from solid tumours was different; particularly in the late S and G 2 + M phases. (author)

  19. Induction of mitosis in the cultured rabbit lens initiated by the addition of insulin to medium KEI-4

    Energy Technology Data Exchange (ETDEWEB)

    Reddan, J R; Unakar, N J; Harding, C V; Bagchi, M; Saldana, G

    1975-01-01

    The epithelium of lenses cultured in KEI-4, a completely defined medium formulated with specific reference to the biochemistry and physiology of the rabbit lens, exhibits a pattern of cell division similar to that noted for the organ in situ. Initial fluctuations in mitotic activity occurred in the area of the germinative zone during the first 24 hr of culture. Mitosis decreased at 1 hr, was extremely low at 3 hr and returned to values comparable for lens in vivo by 22 hr. The precipitous drop in mitosis noted at 3 hr is in part attributable to the isolation of the lens from adjoining tissue. The addition of insulin to KEI-4 triggers a parasynchronous burst of DNA synthesis throughout the central lens epithelium. The activation requires the intact hormone; neither proinsulin nor the A and/or B chains of insulin, nor glucagon nor zinc chloride can initiate mitosis. The gamma-globulin-rich fraction of rabbit serum can also stimulate mitosis. The addition of dibutyryl adenosine 3':5' cyclic monophosphate (DBeAMP) plus theophylline to KEI-4-insulin inhibits mitosis and prevents the cells from entering the synthetic phase of the cell cycle. Theophylline alone or DBeAMP alone brings about a 90 percent reduction in the insulin-induced mitotic responses. Lenses exposed to insulin show a marked increase in RNA synthesis and also exhibit an increased binding of tritiated actinomycin D at 1 and 3 hr of culture relative to KEI-4 controls. The hormone apparently activates the genome including those genes governing cell division. The system is amenable for long-term culture of the mammalian lens and since the constituents of the medium are known it should be possible to determine the factor(s) in the medium which, in conjunction with insulin, are needed for the induction of cell division.

  20. AMMONIA REMOVAL FROM MAMMALIAN CELL CULTURE MEDIUM BY ION-EXCHANGE MEMBRANES

    Science.gov (United States)

    Metabolites such as ammonia and lactic acid formed during mammalian cell culture can frequently be toxic to the cells themselves beyond a threshold concentration of the metabolites. Cell culture conducted in the presence of such accumulated metabolites is therefore limited in pro...

  1. Influence of cell culture medium composition on in vitro dissolution behavior of a fluoride-containing bioactive glass.

    Science.gov (United States)

    Shah, Furqan A; Brauer, Delia S; Wilson, Rory M; Hill, Robert G; Hing, Karin A

    2014-03-01

    Bioactive glasses are used clinically for bone regeneration, and their bioactivity and cell compatibility are often characterized in vitro, using physiologically relevant test solutions. The aim of this study was to show the influence of varying medium characteristics (pH, composition, presence of proteins) on glass dissolution and apatite formation. The dissolution behavior of a fluoride-containing bioactive glass (BG) was investigated over a period of one week in Eagle's Minimal Essential Medium with Earle's Salts (MEM), supplemented with either, (a) acetate buffer, (b) 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, (c) HEPES + carbonate, or (d) HEPES + carbonate + fetal bovine serum. Results show pronounced differences in pH, ion release, and apatite formation over 1 week: Despite its acidic pH (pH 5.8 after BG immersion, as compared to pH 7.4-8.3 for HEPES-containing media), apatite formation was fastest in acetate buffered (HEPES-free) MEM. Presence of carbonate resulted in formation of calcite (calcium carbonate). Presence of serum proteins, on the other hand, delayed apatite formation significantly. These results confirm that the composition and properties of a tissue culture medium are important factors during in vitro experiments and need to be taken into consideration when interpreting results from dissolution or cell culture studies. Copyright © 2013 Wiley Periodicals, Inc.

  2. The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture.

    Science.gov (United States)

    Lee, Jong-Seon; Kim, Ju-Hwan; Seo, Young-Seok; Yang, Jung-Bo; Kim, Yong-Il; Kim, Hye-Jin; Lee, Ki-Hwan

    2013-09-01

    This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos. A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05. The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P media up to 4 weeks did not affect on embryonic development.

  3. Response surface optimization of culture medium for enhanced docosahexaenoic acid production by a Malaysian thraustochytrid.

    Science.gov (United States)

    Manikan, Vidyah; Kalil, Mohd Sahaid; Hamid, Aidil Abdul

    2015-02-27

    Docosahexaenoic acid (DHA, C22:6n-3) plays a vital role in the enhancement of human health, particularly for cognitive, neurological, and visual functions. Marine microalgae, such as members of the genus Aurantiochytrium, are rich in DHA and represent a promising source of omega-3 fatty acids. In this study, levels of glucose, yeast extract, sodium glutamate and sea salt were optimized for enhanced lipid and DHA production by a Malaysian isolate of thraustochytrid, Aurantiochytrium sp. SW1, using response surface methodology (RSM). The optimized medium contained 60 g/L glucose, 2 g/L yeast extract, 24 g/L sodium glutamate and 6 g/L sea salt. This combination produced 17.8 g/L biomass containing 53.9% lipid (9.6 g/L) which contained 44.07% DHA (4.23 g/L). The optimized medium was used in a scale-up run, where a 5 L bench-top bioreactor was employed to verify the applicability of the medium at larger scale. This produced 24.46 g/L biomass containing 38.43% lipid (9.4 g/L), of which 47.87% was DHA (4.5 g/L). The total amount of DHA produced was 25% higher than that produced in the original medium prior to optimization. This result suggests that Aurantiochytrium sp. SW1 could be developed for industrial application as a commercial DHA-producing microorganism.

  4. Medium dependant production of corymbiferone a novel product from Penicillium hordei cultured on plant tissue agar

    DEFF Research Database (Denmark)

    Overy, David Patrick; Zidorn, C.; Petersen, B.O.

    2005-01-01

    Medium dependant production and the structure elucidation of corymbiferone (1) from the fungus Penicillitan hordei grown on oatmeal and macerated tulip, yellow onion and red onion agars are reported. Compound 1 possesses an unusual oxygenated aromatic structure with a lactone bridge preventing full...

  5. Quality tests of culture medium for microbiological analysis of drinkable water

    International Nuclear Information System (INIS)

    Roncoroni, M.

    2000-01-01

    For each of the parameters considered, in accordance with the water's law standards concerning drinking water, they have compared various culture media produced by different manufacturers, in order to choose the products with best selectivity and quality. With this purpose, known microbial cultures (Atcc cultures specified in >) have been used at first, in order to verify the selectivity of soils and the type of growth related with the morphology of the colonies developed. Later, sowings of superficial water of Lake Como have been made, in which a mixed microbial population was present, in order to verify selectivity and possible interference in the execution of an analysis of a real sample [it

  6. Differential Effect of Medium on the Ratio of ICM/TE of Bovine Embryos in a Co-culture System

    Directory of Open Access Journals (Sweden)

    Mohsen Forouzanfar

    2010-01-01

    Full Text Available Background: This study was undertaken to investigate the efficiency of two differentembryo somatic cell co-culture conditions, tissue culture medium 199 (TCM199–vero cellsand Menezo B2 (B2-vero cells, for the in vitro developmental quantity and quality of bovineembryos.Materials and Methods: Bovine oocytes were allowed to mature and subsequently undergofertilization in vitro. Their presumptive zygotes were cultured in either TCM199 or B2 culturemedia in conjunction with vero cells for up to nine days. The culture media were refreshedevery two days and the proportion of embryos that cleaved and further developed to themorula and blastocyst (early, expand and hatched stages were recorded. Hatched blastocystsunderwent differential staining in order to determine the numbers of inner cell mass (ICMand tropho ectoderm (TE and total cell number (TCN.Results: Of the two groups, no significant difference was seen between the proportions ofthe presumptive zygotes cleaved, those which developed to 8-16 cells, morula and reacheddays 7or 8 blastocyst stage or hatched. However, the values for TCN and TE of the TCM199-vero embryos were significantly greater than those of B2-vero embryos. The values for ICM/TCN and ICM/TE were significantly greater in the B2-vero group versus the TCM199-verogroup.Conclusion: Both TCM199 and B2 culture media in conjunction with vero cells were ofthe same efficiency when used for in vitro development of bovine presumptive zygotes.However, TCM199 was superior in providing embryos with more embryo cell numbers,whereas B2 medium was superior in providing embryos with greater ICM/TE and ICM/TCN ratios.

  7. EFFECT OF TREATED DOMESTIC WASTEWATER USED AS CULTURE MEDIUM ON THE GROWTH AND PRODUCTIVITY OF Chlamydomonas sp. STRAIN ISOLATED FROM LANDFILL LEACHATE

    Directory of Open Access Journals (Sweden)

    Fábio de Farias Neves

    2013-07-01

    Full Text Available Microalgae have been culturing to fix carbon and produce biofuels from the biomass. However, it is important to develop low cost strategies for microalgae production in orther to make it a viable alternative of renewable energy. The present research studied the effect of treated wastewater used as an alternative culture medium for growth and productivity of a Chlamydomonas sp. strain isolated from landfills leachate of a treatment pond located in Southern Brazil. Three culture media were evaluated, the control consisted of synthetic TAP medium, other, consisting of 50% TAP medium and 50% wastewater, and another consisting of 100% wastewater. The growth parameters do not have significant difference among the three culture media. Also, productivity do not have significant difference among the cultures with TAP medium and with 100% wastewater, resulting in dry weight values of 1,4±0,14g/L and 1,3±0,19g/L respectively. The culture with 50% TAP medium and 50% wastewater showed the highest productivity, showing an average dry weight value of 1,7±0,07g/L. The results indicate that treated wastewater can be used as an alternative culture medium for Chlamydomonas sp. strain without negative effects on growth and productivity, and possible leading to a decrease in production costs.

  8. Solid tissue culture for cytogenetic analysis: a collaborative survey for the Association of Clinical Cytogeneticists.

    Science.gov (United States)

    Rodgers, C S; Creasy, M R; Fitchett, M; Maliszewska, C T; Pratt, N R; Waters, J J

    1996-01-01

    AIMS: To survey the diagnostic service provided by UK laboratories for the culture of solid tissue samples (excluding tumours) and in particular to examine the variation in culture success rates and the problems of maternal cell overgrowth. METHODS: Twenty seven laboratories took part in a collaborative survey during 1992. Each laboratory submitted data on up to a maximum of 60 consecutive specimens (n = 1361) over a six month period. RESULTS: Skin specimens, the largest category received (n = 520), were the most problematic (51% success rate). Culture success rates were significantly lower (43%) when skin specimens (n = 140) were transported dry to the laboratory. Success rates for skin specimens also varied, depending on the origin of the specimen, from 18% for intra-uterine deaths (IUD) (n = 94) to 85% for neonatal deaths (n = 33) and 83% for live patients (n = 54). Culture of selected extra-fetal tissues from IUD, stillbirths and following elective termination of pregnancy (TOP) gave comparable success rates to those achieved for skin samples from neonatal deaths and live births. Skewed sex ratios, female > male, were identified for products of conception (POC) (n = 298) and placental biopsy specimens (n = 97). CONCLUSIONS: By appropriate selection, transport and processing of tissues, and in particular by avoiding relying solely on skin samples from IUD, stillbirths and TOP, an increase in culture success rates for solid tissue samples submitted for cytogenetic analysis could be achieved. The high risk of maternal cell contamination from POC and placental biopsy specimens was also identified in this survey. PMID:8881913

  9. Development of humanized culture medium with plant-derived serum replacement for human pluripotent stem cells

    Czech Academy of Sciences Publication Activity Database

    Kunová, M.; Matulka, K.; Eiselleová, L.; Trčková, P.; Hampl, Aleš; Dvořák, Petr

    2010-01-01

    Roč. 21, - (2010), s. 676-686 ISSN 1472-6483 Grant - others:GA MŠk(CZ) LC06077; EC FP6(XE) LSHG-CT-2006-018739 Program:LC Institutional research plan: CEZ:AV0Z50390512; CEZ:AV0Z50390703 Keywords : animal protein-free culture * high-density culture * human embryonic stem cells Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.285, year: 2010

  10. Banana peel culture as an indigenous medium for easy identification of late-sporulation human fungal pathogens.

    Science.gov (United States)

    Kindo, A J; Tupaki-Sreepurna, A; Yuvaraj, M

    2016-01-01

    Fungi are increasing in incidence as human pathogens and newer and rarer species are continuously being encountered. Identifying these species from growth on regular culture media may be challenging due to the absence of typical features. An indigenous and cheap medium, similar to the natural substrate of these fungi, was standardised in our laboratory as an aid to species identification in a conventional laboratory setting. Ripe banana peel pieces, sterilised in an autoclave at 121°C temperature and 15 lbs pressure for 15 min promoted good growth of hyphae and pycnidia or acervuli in coelomycetes, flabelliform and medusoid fruiting bodies of basidiomycetes and fruit bodies such as cleistothecium in ascomycetes. The growth from the primary isolation medium was taken and inoculated onto the pieces of double-autoclaved ripe banana peel pieces in a sterile glass Petri dish with some moisture (sprinkles of sterile distilled water). A few sterile coverslips were placed randomly inside the Petri dish for the growing fungus to stick on to it. The plates were kept at room temperature and left undisturbed for 15-20 days. At a time, one coverslip was taken out and placed on a slide with lactophenol cotton blue and focused under the microscope to look for fruit bodies. Lasiodiplodia theobromae, Macrophomina phaseolina, Nigrospora sphaerica, Chaetomium murorum, Nattrassia mangiferae and Schizophyllum commune were identified by characteristic features from growth on banana peel culture. Banana peel culture is a cheap and effective medium resembling the natural substrate of fungi and is useful for promoting characteristic reproductive structures that aid identification.

  11. Evaluation of Epicoccum nigrum for growth, morphology and production of natural colorants in liquid media and on a solid rice medium

    DEFF Research Database (Denmark)

    Mapari, Sameer Shamsuddin; Meyer, Anne S.; Thrane, Ulf

    2008-01-01

    in M1 medium (unoptimized) containing 5 g/l yeast autolysate. The color hue of the crude pigment extracts ranged from 74 to 102 exhibiting dark orange to green-yellow color. Pelleted morphology was shown to have a positive influence on the pigment production by E. nigrum strain IBT 41028 in the liquid...... genera preferred rice but E. nigrum produced 4.6 folds higher pigment in the liquid unoptimized fermentation medium compared to M. ruber. Solid phase extraction and subsequently HPLC-DAD analysis of the crude pigment extracts showed qualitative as well as quantitative variation in the pigment composition...

  12. The availability of a lactose medium for tea fungus culture and Kombucha fermentation

    Directory of Open Access Journals (Sweden)

    Markov S.L.

    2012-01-01

    Full Text Available Kombucha is a traditional beverage that is prepared by fermenting sucrose-sweetened black tea. A medium is inoculated with a cellulose pellicle (popularly known as a “tea fungus” or fermentation brought from previous cultivation process. Our aim was to test the possibility of obtaining a Kombucha beverage using different concentration of lactose as an alternative source of C-atoms. A traditional medium sweetened with sucrose or without sugar was used as control. Without lactose-fermenting yeast strains in tea fungus, lactose is not an adequate alternative source of the C-atom for Kombucha fermentation because it is not possible to obtain Kombucha with an appropriate acidity during a seven-day fermentation. Compared with the traditional medium, fermentation is significantly slower with high differences in acid content. In unsweetened tea inoculated with the beverage obtained from a previous traditional process, Kombucha fermentation processes and produces a beverage without sugar and alcohol. [Projekat Ministarstva nauke Republike Srbije, br. TR 31044

  13. CHROMagar COL-APSE: a selective bacterial culture medium for the isolation and differentiation of colistin-resistant Gram-negative pathogens

    DEFF Research Database (Denmark)

    Abdul Momin, Muhd Haziq F; Bean, David C; Hendriksen, Rene S.

    2017-01-01

    Purpose. A selective chromogenic culture medium for the laboratory isolation and differentiation of colistin resistant Acinetobacter, Pseudomonas, Stenotrophomonas and Enterobacteriaceae spp. (CHROMagar COL-APSE) was developed, evaluated and compared to an existing selective bacterial culture......-resistant non-fermentative bacteria (Acinetobacter, Pseudomonas and Stenotrophomonas). CHROMagar COL-APSE was also more sensitive in supporting the growth of Enterobacteriaceae with COL resistance associated with the carriage of mcr-1. Conclusion. CHROMagar COL-APSE is a sensitive and specific medium...

  14. Chromium (VI) biosorption and removal of chemical oxygen demand by Spirulina platensis from wastewater-supplemented culture medium.

    Science.gov (United States)

    Magro, Clinei D; Deon, Maitê C; De Rossi, Andreia; Reinehr, Christian O; Hemkemeier, Marcelo; Colla, Luciane M

    2012-01-01

    The inappropriate discharge of wastewater containing high concentrations of toxic metals is a serious threat to the environment. Given that the microalga Spirulina platensis has demonstrated a capacity for chromium VI (Cr (VI) biosorption, we assessed the ideal concentration of chromium-containing wastewater required for maximum removal of Cr (VI) and chemical oxygen demand (COD) from the environment by using this microalga. The Paracas and Leb-52 strains of S. platensis, with initial wastewater concentrations of 0%, 12.5%, 25%, and 50%, were cultured in Zarrouk medium diluted to 50% under controlled air, temperature, and lighting conditions. The cultures were maintained for 28 days, and pH, biomass growth, COD, and Cr (VI) were assessed. The wastewater concentration influenced microalgal growth, especially at high concentrations. Removal of 82.19% COD and 60.92% Cr (VI) was obtained, but the COD removal was greater than the Cr (VI) removal in both strains of S. platensis.

  15. Local Roots, Global aspirations: Impact of culture on work environment and organizational culture in Malaysian Small and Medium Enterprises in the Information Technology Sector

    Directory of Open Access Journals (Sweden)

    Saxena Vandana

    2017-01-01

    Full Text Available This paper investigates the role of culture in hiring, team formations and workplace interactions in Malaysian small and medium enterprises (SME in the Information and Communication Technology (ICT sector. This research used the case study approach, with multimethod data collecting instruments like observation, interviews, and analysis of the data available on the websites of the two ICT SMEs under study. The participants selected for the study were the owners, managers and senior employees of both firms. While both firms operated in similar fields, the workforce of one consisted largely of Malaysian employees, while that of second company consisted largely of foreigners. The findings revealed a considerable bias and preference towards cultural homogeneity.

  16. In vitro culture and medium-term conservation of the rare wild ...

    African Journals Online (AJOL)

    Yomi

    Initiation of in vitro cultures was started from wounded seeds or excised mature embryos. ... the South Scandinavian hills to the coast of. Mediterranean Sea but its origin is considered to be East. Europe and ... It belongs to Euro-Asian species and ..... Indian. Hort. J. 48:171-178. Rakosy-Tican L, Balogh A, Copaci C (2000).

  17. The Contribution of Cultural Capital to Students' Mathematics Achievement in Medium and High Socioeconomic Gradient Economies

    Science.gov (United States)

    Tan, Cheng Yong

    2015-01-01

    The present study addresses the issue of how different forms of cultural capital may influence children's mathematics achievement in economies with different socioeconomic gradients. Data from 73,178 parent-child dyads from 10 economies with different socioeconomic gradients who participated in the Programme for International Student Assessment…

  18. Evidence of biogenic corrosion of titanium after exposure to a continuous culture of thiobacillus ferrooxidans grown in thiosulfate medium

    International Nuclear Information System (INIS)

    Horn, J M; Martin, S I; Masterson, B

    2000-01-01

    Experiments were undertaken to evaluate extreme conditions under which candidate materials intended for use in a proposed nuclear waste repository might be susceptible to corrosion by endogenous microorganisms. Thiobucillus ferrooxidans, a sulfur-oxidizing bacterium, was grown in continuous culture using thiosulfate as an energy source; thiosulfate is oxidized to sulfate as a metabolic endproduct by this organism. Culture conditions were optimized to produce a high-density, metabolically active culture throughout a period of long term incubation in the presence of Alloy 22 (a high nickel-based alloy) and Titanium grade 7 (Tigr7) material coupons. After seven months incubation under these conditions, material coupons were withdrawn and analyzed by high resolution microscopy and energy dispersive x-ray analyses. Alloy 22 coupons showed no detectable signs of corrosion. Tigr7, however, demonstrated distinct roughening of the coupon surface, and [presumably solubilized and precipitated] titanium was detected on Alloy 22 coupons incubated in the same T. ferrooxiduns culture vessel. Control coupons of these materials incubated in sterile thiosulfate medium did not demonstrate any signs of corrosion, thus showing that observed corrosive effects were due to the T. ferrooxidans metabolic activities. T. ferrooxidans intermediates of thiosulfate oxidation or sulfate may have caused the corrosive effects observed on Tigr7

  19. The Effect of Medium Cultures on Water Use and Charactristic of Gazania Flowers (Gazania hybrida in Green Roof.

    Directory of Open Access Journals (Sweden)

    Tahereh Bahrami

    2017-09-01

    Full Text Available Introduction: Green roof is one of the newest phenomenons in architecture and urbanism that refers to the sustainable development concepts and it will be usable for increasing landscape design, improving quality of the environment and reduction in energy consumption. Ensure of existing adequate green landscape in urban areas and improving access to natural areas surrounding the cities can help to offset negative effects of urban life. The use of green roof technology in cities is one of advanced techniques of green landscape. A green or living roof is a roof of a building that is partially or completely covered with vegetation and a growing medium on top view of buildings. Green roof layers that considered for roof side consist of protection layer, drainage layer, growing medium and plant layer. Medium layer is the medium culture of green roof that plants are begins to grow in it. This space should enable to save enough minerals and water for conserve of green-roof plants. All kinds of plants can growth on the green roof, but there are some constraints in creative of design because of roots dimension, plant canopy, necessary volume of soil, suitable direction to light, good weather, weight of designed structures, budget of repairing and keeping. Materials and Methods: To evaluate the effect of some culture medium on water consumption, vegetative and reproductive traits of Gazania (Gazania hybrida in condition of green roof a factorial experiment was conducted based on a completely randomized design with nine treatments and three replications in 2014. Treatments were three levels of vermicompost (zero, 5%, and 10% and rice hull (zero, 7, and 14%. Seedlings of plants cultivated in the media mixture of coco peat 15%, perlite 15%, leaf 10%, manure 10%, and filed soil 50%. The container had 60 × 60 ×25 cm dimensions that placed on the roof of greenhouse building with four meters height. The measured traits was number, average, and diameter of

  20. The Effect of Medium Cultures on Water Use and Charactristic of Gazania Flowers (Gazania hybrida in Green Roof.

    Directory of Open Access Journals (Sweden)

    Tahereh Bahrami

    2017-02-01

    Full Text Available Introduction: Green roof is one of the newest phenomenons in architecture and urbanism that refers to the sustainable development concepts and it will be usable for increasing landscape design, improving quality of the environment and reduction in energy consumption. Ensure of existing adequate green landscape in urban areas and improving access to natural areas surrounding the cities can help to offset negative effects of urban life. The use of green roof technology in cities is one of advanced techniques of green landscape. A green or living roof is a roof of a building that is partially or completely covered with vegetation and a growing medium on top view of buildings. Green roof layers that considered for roof side consist of protection layer, drainage layer, growing medium and plant layer. Medium layer is the medium culture of green roof that plants are begins to grow in it. This space should enable to save enough minerals and water for conserve of green-roof plants. All kinds of plants can growth on the green roof, but there are some constraints in creative of design because of roots dimension, plant canopy, necessary volume of soil, suitable direction to light, good weather, weight of designed structures, budget of repairing and keeping. Materials and Methods: To evaluate the effect of some culture medium on water consumption, vegetative and reproductive traits of Gazania (Gazania hybrida in condition of green roof a factorial experiment was conducted based on a completely randomized design with nine treatments and three replications in 2014. Treatments were three levels of vermicompost (zero, 5%, and 10% and rice hull (zero, 7, and 14%. Seedlings of plants cultivated in the media mixture of coco peat 15%, perlite 15%, leaf 10%, manure 10%, and filed soil 50%. The container had 60 × 60 ×25 cm dimensions that placed on the roof of greenhouse building with four meters height. The measured traits was number, average, and diameter of

  1. Effect of volume ratio of liquid to solid on the interfacial microstructure and mechanical properties of high chromium cast iron and medium carbon steel bimetal

    International Nuclear Information System (INIS)

    Xiong Bowen; Cai Changchun; Lu Baiping

    2011-01-01

    Highlights: → Volume ratio of liquid to solid affects significantly the interfacial microstructure. → Elemental diffusion activity is increased by increasing volume ratio. → Mechanical property is improved by increasing volume ratio. - Abstract: The high chromium cast iron and medium carbon steel bimetal was fabricated by liquid-solid casting technology. The effect of volume ratios of liquid to solid (6:1, 10:1 and 12:1) on the interfacial microstructure and mechanical properties of bimetal was investigated. The interfacial microstructure was analyzed using scanning electron microscope (SEM) and transmission electron microscope (TEM). The shear strength and microhardness in as-cast condition were studied at room temperature. The results show that the volume ratios of liquid to solid affect significantly the interfacial microstructure. When liquid-solid volume ratio was 6:1, the unbonded region was detected in interface region because the imported heat energy cannot support effectively the diffusion of element, whereas, when liquid-solid volume ratios reach 10:1 and 12:1, a sound interfacial microstructure was achieved by the diffusion of C, Cr, Mo, Cu and Mn, and metallurgical bonding without unbonded region, void and hole, etc. was detected. With the increase of liquid-solid volume ratio, the elemental diffusion activity improves, resulting in the increase of width of interface transition region. At the same distance from interface, with the increase of liquid-solid volume ratio, the microhardness is degraded in HCCI, but increased in MCS. The shear strength is also improved with the increase of liquid-solid volume ratio.

  2. Preirradiation of medium induces a subsequent stimulation or inhibition of growth according to the physiological state in Synechococcus lividus in culture

    International Nuclear Information System (INIS)

    Conter, A.

    1987-01-01

    The proliferation of Synechococcus lividus cells grown in preirradiated medium was compared with the proliferation of cells grown in a shielded or freshly prepared medium. Aging of medium in a shielded chamber resulted in a slight inhibiting effect on growth in every phase of the cell cycle which was used. Preirradiation of medium resulted in a stimulation of growth observed on Day 7 in cultures inoculated with cells selected in the deceleration phase and an inhibition of growth in cultures inoculated with exponentially growing cells. Addition of catalase (100 U X ml-1) counteracted the stimulating effect but did not modify the inhibiting effect induced by preirradiated medium. Results demonstrated the indirect effect of low doses of irradiation, implying the presence of hydrogen peroxide in radiostimulation and other radioproducts in the inhibitory effect

  3. AMC-Bio-Artificial Liver culturing enhances mitochondrial biogenesis in human liver cell lines: The role of oxygen, medium perfusion and 3D configuration

    NARCIS (Netherlands)

    Adam, Aziza A. A.; van Wenum, Martien; van der Mark, Vincent A.; Jongejan, Aldo; Moerland, Perry D.; Houtkooper, Riekelt H.; Wanders, Ronald J. A.; Oude Elferink, Ronald P.; Chamuleau, Robert A. F. M.; Hoekstra, Ruurdtje

    2017-01-01

    Human liver cell lines, like HepaRG and C3A, acquire higher functionality when cultured in the AMC-Bio-Artificial Liver (AMC-BAL). The three main differences between BAL and monolayer culture are the oxygenation (40% vs 20%O2), dynamic vs absent medium perfusion and 3D vs 2D configuration. Here, we

  4. The conversion of BTEX compounds by single and defined mixed cultures to medium-chain-length polyhydroxyalkanoate.

    Science.gov (United States)

    Nikodinovic, Jasmina; Kenny, Shane T; Babu, Ramesh P; Woods, Trevor; Blau, Werner J; O'Connor, Kevin E

    2008-09-01

    Here, we report the use of petrochemical aromatic hydrocarbons as a feedstock for the biotechnological conversion into valuable biodegradable plastic polymers--polyhydroxyalkanoates (PHAs). We assessed the ability of the known Pseudomonas putida species that are able to utilize benzene, toluene, ethylbenzene, p-xylene (BTEX) compounds as a sole carbon and energy source for their ability to produce PHA from the single substrates. P. putida F1 is able to accumulate medium-chain-length (mcl) PHA when supplied with toluene, benzene, or ethylbenzene. P. putida mt-2 accumulates mcl-PHA when supplied with toluene or p-xylene. The highest level of PHA accumulated by cultures in shake flask was 26% cell dry weight for P. putida mt-2 supplied with p-xylene. A synthetic mixture of benzene, toluene, ethylbenzene, p-xylene, and styrene (BTEXS) which mimics the aromatic fraction of mixed plastic pyrolysis oil was supplied to a defined mixed culture of P. putida F1, mt-2, and CA-3 in the shake flasks and fermentation experiments. PHA was accumulated to 24% and to 36% of the cell dry weight of the shake flask and fermentation grown cultures respectively. In addition a three-fold higher cell density was achieved with the mixed culture grown in the bioreactor compared to shake flask experiments. A run in the 5-l fermentor resulted in the utilization of 59.6 g (67.5 ml) of the BTEXS mixture and the production of 6 g of mcl-PHA. The monomer composition of PHA accumulated by the mixed culture was the same as that accumulated by single strains supplied with single substrates with 3-hydroxydecanoic acid occurring as the predominant monomer. The purified polymer was partially crystalline with an average molecular weight of 86.9 kDa. It has a thermal degradation temperature of 350 degrees C and a glass transition temperature of -48.5 degrees C.

  5. The Effect of Fermentation Time with Probiotic Bacteria on Organic Fertilizer as Daphnia magna Cultured Medium towards Nutrient Quality, Biomass Production and Growth Performance Enhancement

    Science.gov (United States)

    Endar Herawati, Vivi; Agung Nugroho, Ristiawan; Pinandoyo; Darmanto, YS; Hutabarat, Johannes

    2018-02-01

    The nutrient quality and growth performance of D. magna are highly depend on the organic fertilizer which is used in its culture medium. The objective of this study was to identify the best fermentation time by using probiotic bacteria on organic fertilizer as mass culture medium to improve its nutrient quality, biomass production, and growth performance. This study was conducted using completely randomized experimental design with five treatments and three repetitions. Organic fertilizers used cultured medium with chicken manure, rejected bread and tofu waste fermented by probiotic bacteria then cultured for 0, 7, 14, 21 and 28 days. The results showed that medium which used 25% chicken manure, 25% tofu waste and 50% rejected bread cultured for 28 days created the highest biomass production, population density and nutrient content of D. magna those are 233,980 ind/L for population density; 134.60 grams for biomass production, 0.574% specific growth rate; 68.06% protein content and 6.91% fat. The highest fatty acid profile is 4.83% linoleic and 3.54% linolenic acid. The highest essential amino acid is 53.94 ppm lysine. In general, the content of ammonia, DO, temperature, and pH during the study were in the good range of D. magna life. The conclusion of this research is medium which used 25% chicken manure, 25% tofu waste and 50% rejected bread cultured for 28 days created the highest biomass production, population and nutrient content of D. magna.

  6. Influence of culture conditions and medium composition on the production of antibacterial compounds by marine Serratia sp. WPRA3.

    Science.gov (United States)

    Jafarzade, Mahtab; Yahya, Nur Ain; Shayesteh, Fatemeh; Usup, Gires; Ahmad, Asmat

    2013-06-01

    This study was undertaken to investigate the influence of culture conditions and medium components on production of antibacterial compounds by Serratia sp. WPRA3 (JX020764) which was isolated from marine water of Port Dickson, Malaysia. Biochemical, morphological, and molecular characteristics suggested that the isolate is a new candidate of the Serratia sp. The isolate showed strong antimicrobial activity against fungi, Gram-negative and Gram-positive bacteria. This bacterium exhibited optimum antibacterial compounds production at 28°C, pH 7 and 200 rev/min aeration during 72 h of incubation period. Highest antibacterial activity was obtained when sodium chloride (2%), yeast extract (0.5%), and glucose concentration (0.75%) were used as salt, nitrogen, and carbon sources respectively. Different active fractions were obtained by Thin-Layer Chromatography (TLC) and Flash Column Chromatography (FCC) from ethyl acetate crude extracts namely OCE and RCE in different culture conditions, OCE (pH 5, 200 rev/min) and RCE (pH 7/without aeration). In conclusion, the results suggested different culture conditions have a significant impact on the types of secondary metabolites produced by the bacterium.

  7. Effect of the double mutant e//e w//w and the culture medium on the productivity of Drosophila melanogaster

    Directory of Open Access Journals (Sweden)

    Francisco Mora

    2000-01-01

    Full Text Available We investigated the effect of two culture media on the productivity of the double mutant ebony-white (e//e w//w of Drosophila melanogaster, aimed at improving the conditions for maintenance of Drosophila’s collection, Departamento de Biología, Universidad Nacional de Colombia. The results indicate that the productivity is affected by the culture medium, being the maize culture medium more productive than the wheat one; it was also shown that the productivity depends both, on the crosses type that is realize and on the mutant. The “+//+ +//+ x e//e w/ cross is more productive than its reciprocal cross, where the position of the ebony allele is the most important factor. With respect to the white allele, when carried by males it does not have effect on the productivity. In addition, we detected a negative effect of wheat culture medium on females +//e +//w.

  8. Enhanced production of nisin by co-culture of Lactococcus lactis sub sp. lactis and Yarrowia lipolytica in molasses based medium.

    Science.gov (United States)

    Ariana, Mehdi; Hamedi, Javad

    2017-08-20

    Nisin is a safe, approved and commercial bacteriocin that is produced by Lactococcus lactis subsp. lactis. Since lactate accumulation in fermentation medium reduces L. lactis growth and nisin production, Yarrowia lipolytica, a lactate consuming yeast and L. lactis subsp. lactis, were simultaneously cultured in a molasses based medium. Y. lipolytica is not able to consume sucrose as carbon source, but rather consumes lactate and hence decrease lactic acid titer by 10% in the medium. Lactic acid consumption, 15% increased pH value and stimulated L. lactis growth. In the mixed culture, nisin production and L. lactis growth were 50% and 49% higher than that of pure culture, respectively. Also the results showed that specific growth rate of L. lactis increased 4 times more than that of the pure culture. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Production of Medium Chain Length Polyhydroxyalkanoates From Oleic Acid Using Pseudomonas putida PGA1 by Fed Batch Culture

    Directory of Open Access Journals (Sweden)

    Sidik Marsudi

    2010-10-01

    Full Text Available Bacterial polyhydroxyalkanoates (PHAs are a class of p0lymers currently receiving much attention because of their potential as renewable and biodegradable plastics. A wide variety of bacteria has been reported to produce PHAs including Pseudomonas strains. These strains are known as versatile medium chain length PHAs (PHAs-mcl producers using fatty acids as carbon source. Oleic acid was used to produce PHAs-mcl using Pseudomonas putida PGA 1 by continuous feeding of both nitrogen and carbon source, in a fed batch culture. During cell growth, PHAs also accumulated, indicating that PHA production in this organism is growth associated. Residual cell increased until the nitrogen source was depleted. At the end of fermentation, final cell concentration, PHA content, and roductivity were 30.2 g/L, 44.8 % of cell dry weight, and 0.188 g/l/h, respectively.

  10. The Spiritual Form of Ancient Art and Culture - Bharatanatyam (Visual Art Depicted Using Unique Techniques on Scratchboard (Fine Art Medium

    Directory of Open Access Journals (Sweden)

    Arpitha Parthasarathy

    2017-03-01

    Full Text Available The most ancient form of dance that is prevailing todays is a form of classical Indian dance, Bharatanatyam. In Sanskrit (and Devanagri, bharatanatyam means "Indian dance", is believed to have divine origin and is of the most ancient form of classical dance. Bharatanatyam is a two thousand-year-old dance form, originally practiced in the temples of ancient India. The art today remains purely devotional even today and this performing art is yet to gain awareness and interest in the western world. This dance form has various implications in improving the higher order thinking in children and provides health benefits in adults apart from cultural preservation. The current study uses scratchboard as a medium to display the artistic movements and emotions. Scratchboard, a fine art is one means by which the visual art is expressed in this current study using sharp tools, namely X-acto 11 scalpel and tattoo needles. This unique medium made up of a masonite hardboard coated with soft clay and Indian ink has been used to not only show the details of the ancient dance form and expression but also to comprehend and transcribe both visual art and fine art. It is for the first time that scratchboard medium has been the innovatively used to show various textures of flower, glistening gold jewels, hand woven silk and the divine expression in the same art ‘devotion’. The current study was carried out in-order to perpetuate, conserve and disseminate these classic forms of visual art and fine art.

  11. Combined data preprocessing and multivariate statistical analysis characterizes fed-batch culture of mouse hybridoma cells for rational medium design.

    Science.gov (United States)

    Selvarasu, Suresh; Kim, Do Yun; Karimi, Iftekhar A; Lee, Dong-Yup

    2010-10-01

    We present an integrated framework for characterizing fed-batch cultures of mouse hybridoma cells producing monoclonal antibody (mAb). This framework systematically combines data preprocessing, elemental balancing and statistical analysis technique. Initially, specific rates of cell growth, glucose/amino acid consumptions and mAb/metabolite productions were calculated via curve fitting using logistic equations, with subsequent elemental balancing of the preprocessed data indicating the presence of experimental measurement errors. Multivariate statistical analysis was then employed to understand physiological characteristics of the cellular system. The results from principal component analysis (PCA) revealed three major clusters of amino acids with similar trends in their consumption profiles: (i) arginine, threonine and serine, (ii) glycine, tyrosine, phenylalanine, methionine, histidine and asparagine, and (iii) lysine, valine and isoleucine. Further analysis using partial least square (PLS) regression identified key amino acids which were positively or negatively correlated with the cell growth, mAb production and the generation of lactate and ammonia. Based on these results, the optimal concentrations of key amino acids in the feed medium can be inferred, potentially leading to an increase in cell viability and productivity, as well as a decrease in toxic waste production. The study demonstrated how the current methodological framework using multivariate statistical analysis techniques can serve as a potential tool for deriving rational medium design strategies. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. Modification of growth medium of mixed-culture species of microalgae isolated from southern java coastal region

    Directory of Open Access Journals (Sweden)

    Sudibyo Hanifrahmawan

    2018-01-01

    Full Text Available Globally, there is growing interest in microalgae as production organisms. Microalgae contain lipids (oil, proteins and carbohydrates (sugars, and, especially marine algae have been used as food and feed for centuries. Recently, production cost reduction related to the supply of growth nutrients is necessary to make it profitable. Therefore, utilization of molasses, a byproduct of sugar production, as the natural carbon, macronutrients, and micronutrients sources can be helpful. The analysis showed that the content of sucrose, glucose, fructose, potassium, zinc, and magnesium was 68.4% w/w, 18.5% w/w, and 13.1% w/w, 5.5% w/w, 3.91 ppm, and 1,370 ppm respectively. This work aimed to determine the effect of molasses addition to the physio-chemical properties of multi-culture species of microalgae isolated from southern Java coastal region in Indonesia grown under mixotrophic culture. The cultivation in this work used medium which was self-formulated by the authors consisting of NaNO3 (5 mL/L, H3BO3 (1 mL/L, EDTA (1 mL/L, N2H2PO4 (5 mL/L, FeSO4 (1 mL/L, MgSO4 (1 mL/L, NaCl (1 mL/L, micronutrients (1 mL/L, vitamin B1 (1 mL/L, and vitamin B12 (1 mL/L in 500 mL of water. The medium will be treated to have molasses concentration of 0.05% v/v, 0.15% v/v, 0.25% v/v, 0.35% v/v, and 0.45% v/v.

  13. Studies on the effects of phosphine on Salmonella enterica serotype Enteritidis in culture medium and in black pepper (Piper nigrum).

    Science.gov (United States)

    Castro, M F P M; Rezende, A C B; Benato, E A; Valentini, S R T; Furlani, R P Z; Tfouni, S A V

    2011-04-01

    The effect of phosphine on Salmonella enterica serotype Enteritidis inoculated in culture medium and in black pepper grains (Piper nigrum), as well as on the reduction of the microbial load of the dried and moisturized product, was verified. The postfumigation effect was verified in inoculated samples with 0.92 and 0.97 water activity (a(w)) exposed to 6 g/m(3) phosphine for 72 h, dried to 0.67 a(w), and stored for 24, 48, and 72 h. No decreases were observed in Salmonella Enteritidis populations in culture medium when fumigant concentrations up to 6 g/m(3) were applied for 48 h at 35°C. However, the colonies showed reductions in size and atypical coloration as the phosphine concentration increased. No reduction in Salmonella counts occurred on the inoculated dried samples after fumigation. On the other hand, when phosphine at concentrations of 6 g/m(3) was applied on moisturized black pepper for 72 h, decreases in Salmonella counts of around 80% were observed. The counts of total aerobic mesophilic bacterium populations of the dried and moisturized black pepper were not affected by the fumigant treatment. The results of the postfumigation studies indicated that Salmonella Enteritidis was absent in the fumigated grains after drying and storage for 72 h, indicating a promising application for this technique. It was concluded that for Salmonella Enteritidis control, phosphine fumigation could be applied to black pepper grains before drying and the producers should rigidly follow good agricultural practices, mainly during the drying process, in order to avoid product recontamination. Additional work is needed to confirm the findings with more Salmonella serotypes and strains.

  14. A simple, specific high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin in cell culture medium.

    Science.gov (United States)

    Li, Ye; Cassone, Vincent M

    2015-09-01

    A simple, specific, high-throughput enzyme-linked immunosorbent assay (ELISA) for quantitative determination of melatonin was developed for directly measuring melatonin in cell culture medium with 10% FBS. This assay adopts a commercial monoclonal melatonin antibody and melatonin-HRP conjugate, so it can be applied in multiple labs rapidly with low cost compared with commercial RIA and ELISA kits. In addition, the procedure is much simpler with only four steps: 1) sample/conjugate incubation, 2) plate washing, 3) TMB color reaction and 4) reading of results. The standards of the assay cover a wide working range from 100 pg/mL to 10 ng/mL. The sensitivity was 68 pg/mL in cell culture medium with 10% FBS and 26 pg/mL in PBS with as little as 25 μL sample volume. The recovery of melatonin from cell culture medium was 101.0%. The principal cross-reacting compound was 5-methoxytryptophol (0.1%). The variation coefficients of the assay, within and between runs, ranged between 6.68% and 15.76% in cell culture medium. The mean linearity of a series diluted cell culture medium sample was 105% (CV=5%), ranging between 98% and 111%, y=5.5263x+0.0646, R(2)=0.99. The assay enables small research and teaching labs to reliably measure this important neurohormone. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Improvement of Medium Chain Fatty Acid Content and Antimicrobial Activity of Coconut Oil via Solid-State Fermentation Using a Malaysian Geotrichum candidum

    Directory of Open Access Journals (Sweden)

    Anahita Khoramnia

    2013-01-01

    Full Text Available Coconut oil is a rich source of beneficial medium chain fatty acids (MCFAs particularly lauric acid. In this study, the oil was modified into a value-added product using direct modification of substrate through fermentation (DIMOSFER method. A coconut-based and coconut-oil-added solid-state cultivation using a Malaysian lipolytic Geotrichum candidum was used to convert the coconut oil into MCFAs-rich oil. Chemical characteristics of the modified coconut oils (MCOs considering total medium chain glyceride esters were compared to those of the normal coconut oil using ELSD-RP-HPLC. Optimum amount of coconut oil hydrolysis was achieved at 29% moisture content and 10.14% oil content after 9 days of incubation, where the quantitative amounts of the modified coconut oil and MCFA were 0.330 mL/g of solid media (76.5% bioconversion and 0.175 mL/g of solid media (53% of the MCO, respectively. MCOs demonstrated improved antibacterial activity mostly due to the presence of free lauric acid. The highest MCFAs-rich coconut oil revealed as much as 90% and 80% antibacterial activities against Staphylococcus aureus and Escherichia coli, respectively. The results of the study showed that DIMOSFER by a local lipolytic G. candidum can be used to produce MCFAs as natural, effective, and safe antimicrobial agent. The produced MCOs and MCFAs could be further applied in food and pharmaceutical industries.

  16. Improvement of medium chain fatty acid content and antimicrobial activity of coconut oil via solid-state fermentation using a Malaysian Geotrichum candidum.

    Science.gov (United States)

    Khoramnia, Anahita; Ebrahimpour, Afshin; Ghanbari, Raheleh; Ajdari, Zahra; Lai, Oi-Ming

    2013-01-01

    Coconut oil is a rich source of beneficial medium chain fatty acids (MCFAs) particularly lauric acid. In this study, the oil was modified into a value-added product using direct modification of substrate through fermentation (DIMOSFER) method. A coconut-based and coconut-oil-added solid-state cultivation using a Malaysian lipolytic Geotrichum candidum was used to convert the coconut oil into MCFAs-rich oil. Chemical characteristics of the modified coconut oils (MCOs) considering total medium chain glyceride esters were compared to those of the normal coconut oil using ELSD-RP-HPLC. Optimum amount of coconut oil hydrolysis was achieved at 29% moisture content and 10.14% oil content after 9 days of incubation, where the quantitative amounts of the modified coconut oil and MCFA were 0.330 mL/g of solid media (76.5% bioconversion) and 0.175 mL/g of solid media (53% of the MCO), respectively. MCOs demonstrated improved antibacterial activity mostly due to the presence of free lauric acid. The highest MCFAs-rich coconut oil revealed as much as 90% and 80% antibacterial activities against Staphylococcus aureus and Escherichia coli, respectively. The results of the study showed that DIMOSFER by a local lipolytic G. candidum can be used to produce MCFAs as natural, effective, and safe antimicrobial agent. The produced MCOs and MCFAs could be further applied in food and pharmaceutical industries.

  17. Subtractive cloning of cDNA from Aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) culture.

    Science.gov (United States)

    Akao, Takeshi; Gomi, Katsuya; Goto, Kuniyasu; Okazaki, Naoto; Akita, Osamu

    2002-07-01

    In solid-state cultures (SC), Aspergillus oryzae shows characteristics such as high-level production and secretion of enzymes and hyphal differentiation with asexual development which are absent in liquid (submerged) culture (LC). It was predicted that many of the genes involved in the characteristics of A. oryzae in SC are differentially expressed between SC and LC. We generated two subtracted cDNA libraries with bi-directional cDNA subtractive hybridizations to isolate and identify such genes. Among them, we identified genes upregulated in or specific to SC, such as the AOS ( A. oryzae SC-specific gene) series, and those downregulated or not expressed in SC, such as the AOL ( A. oryzae LC-specific) series. Sequencing analyses revealed that the AOS series and the AOL series contain genes encoding extra- and intracellular enzymes and transport proteins. However, half were functionally unclassified by nucleotide sequences. Also, by expression profile, the AOS series comprised two groups. These gene products' molecular functions and physiological roles in SC await further investigation.

  18. Influence of culture medium pH on charasome development and chloride transport in Chara corallina

    International Nuclear Information System (INIS)

    Lucas, W.J.; Keifer, D.W.; Pesacreta, T.C.

    1986-01-01

    Internodal cells of Chara, grown in culture either at pH 5.7,6.5 or 7.5, were studied to determine their chloride influx capability, the quantitative aspects of charasome morphology and the degree to which these two parameters could be correlated. In cells grown at pH 5.7 the charasomes were relatively small, were widely spaced on the plasma membrane, and contributed only at 0.6% increase to the surface area of the plasma membrane in the acid region of the cell. In contrast, the charasome membrane surface area of cells grown at pH 7.5 had increased x 19, the density of charasomes on the cell surface increased x 42, thus producing a x 3.57 increase in the acid region plasma membrane surface area. Chloride influx in cells grown at pH 7.5 was x 8.7-12.7 greater than in cells grown at pH 5.7. Cells that had been starved of chloride exhibited a x 2.4 average increase in the rate of chloride influx. Our observations establish the existence of a positive correlation between the rate of chloride influx and the increase in membran surface area due to charasomes, although other factors, such as the effect of pH on transport-related enzymes, and the effect of charasome structure on chemical equilibria, may also be of importance. (Author)

  19. Biodegradation of Palm Kernel Cake by Cellulolytic and Hemicellulolytic Bacterial Cultures through Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Mohamed Idris Alshelmani

    2014-01-01

    Full Text Available Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ and the American Type Culture Collection (ATCC. Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF of palm kernel cake (PKC, whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly P<0.05 decreased, whereas the amount of reducing sugars were significantly P<0.05 increased in the fermented PKC (FPKC compared with untreated PKC.

  20. Influence of Culture Medium Composition and Light Conditions on the Accumulation of Bioactive Compounds in Shoot Cultures of Scutellaria lateriflora L. (American Skullcap) Grown In Vitro.

    Science.gov (United States)

    Kawka, Beata; Kwiecień, Inga; Ekiert, Halina

    2017-12-01

    Methanolic extracts from in vitro grown Scutellaria lateriflora shoots cultured on five Murashige and Skoog (MS) medium variants supplemented with different combinations of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) under different light conditions (monochromatic light, white light and no light) were analysed by HPLC for three groups of metabolites: flavonoids (26 compounds), phenolic acids and their precursors (19+2) and phenylethanoid glycosides (2). The analyses revealed the presence of baicalein, baicalin, wogonin, wogonoside, 3,4-dihydroxyphenylacetic acid and verbascoside. There was clear evidence of the influence of plant growth regulators and light conditions on the accumulation of the analysed groups of secondary metabolites. The amounts of the compounds changed within a wide range-for the total flavonoid content, 30.2-fold (max. 1204.3 mg·100 g -1 dry weight (DW)); for 3,4-dihydroxyphenylacetic acid, 5.5-fold (max. 33.56 mg·100 g -1 DW); and for verbascoside, 1.5-fold (169.15 max. mg·100 g -1 DW). The best medium for the production of most of the compounds was the Murashige and Skoog variant with 1 mg l -1 BAP and 1 mg l -1 NAA. For verbascoside, the best 'productive' medium was the MS variant supplemented with 0.5 mg l -1 BAP and 2 mg l -1 NAA. The accumulation of the metabolites was stimulated to the greatest extent by blue light, under which the extracts were found to contain the highest total amount of flavonoids and the highest amounts of flavonoid glucuronides, baicalin and wogonoside, as well as of verbascoside. Their amounts were, respectively, 1.54-, 1.49-, 2.05- and 1.86-fold higher than under the control white light.

  1. Magnetic field action on outdoor and indoor cultures of Spirulina: Evaluation of growth, medium consumption and protein profile.

    Science.gov (United States)

    Deamici, Kricelle Mosquera; Santos, Lucielen Oliveira; Costa, Jorge Alberto Vieira

    2018-02-01

    This study aimed at evaluating whether a magnetic field (MF) affects the growth of Spirulina sp. when applied to it at different exposure times in indoor and outdoor culture systems. The effects of MF on chlorophyll content, medium consumption and protein profile were also investigated. In raceway tanks, a 25 mT MF was applied for 24 h or for 1 h d -1 . MF for 24 h to outdoor assays increased biomass concentration and chlorophyll-a content besides altering the protein profile. Outdoor Spirulina growth was higher (∼3.65 g L -1 ) than the growth found in indoor assays (∼1.80 g L -1 ), while nitrogen and phosphorus consumption was not enhanced by the application of MF. This is the first study that investigated the influence of MF on outdoor microalga assays, and the results showed that MF affected the metabolism of Spirulina cultured in raceways, especially when it was grown outdoors in uncontrolled environmental conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Ability of a Lactobacillus rhamnosus strain cultured in milk whey based medium to bind aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Fernanda Bovo

    2014-09-01

    Full Text Available This study aimed to compare Lactobacillus rhamnosus growth in MRS (de Man, Rogosa and Sharpe broth and a culture medium containing milk whey (MMW and to evaluate aflatoxin B1 (AFB1 adsorption capacity by bacterial cells produced in both culture media. L. rhamnosus cells were cultivated in MRS broth and MMW (37 °C, 24 hours, and bacterial cell concentration was determined spectrophotometrically at 600 nm. AFB1 (1 µg/ml adsorption assays were conducted using 1 x 10(10 non-viable L. rhamnosus cells (121 °C, 15 minutes at pHs 3.0 and 6.0 and contact time of 60 minutes. AFB1 quantification was performed by High Performance Liquid Chromatography. Bacterial cell concentration in MMW was higher (9.84 log CFU/ml than that in MRS broth (9.63 log CFU/ml. There were no significant differences between AFB1 binding results at the same pH value (3.0 or 6.0 for the cells cultivated in MRS broth (46.0% and 35.8%, respectively and in MMW (43.7% and 25.8%, respectively, showing that MMW can adequately replace the MRS broth. Therefore, it can be concluded that the use of L. rhamnosus cells cultivated in MMW offers advantages such as reduction in large scale production costs, improvement of environmental sustainability, and being a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.

  3. Solid-stemmed spring wheat cultivars give better androgenic response than hollow-stemmed cultivars in anther culture.

    Science.gov (United States)

    Weigt, Dorota; Kiel, Angelika; Nawracała, Jerzy; Pluta, Mateusz; Łacka, Agnieszka

    2016-01-01

    Solid-stemmed spring wheat cultivars ( Triticum aestivum L.) are resistant to the stem sawfly ( Cephus cinctus Nort.) and lodging. Anthers of 24 spring wheat cultivars with varying content of pith in the stem were used in the experiment. All were classified into three groups: solid, medium-solid and hollow stems. There was considerable influence of the cultivar on callus formation and green plant regeneration. The highest efficiency of green plant regeneration (24%) was observed for the solid-stemmed AC Abbey cultivar. There was no regeneration from the explants of four cultivars: CLTR 7027, Alentejano, Marquis and Bombona. Principal component analysis showed no differences between the cases under observation (callus induction and green plant regeneration) in their response to pre-treatment temperatures (4 and 8°C). The examination of the effects of various auxin types in the induction medium on callus formation and green plant regeneration revealed that the strongest stimulation of these processes was observed in the C17 medium with 2,4-D and dicamba. The efficiency of callus formation and green plant regeneration was greater in solid-stemmed cultivars than in hollow-stemmed cultivars.

  4. Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos.

    Science.gov (United States)

    Arias, María E; Ross, Pablo J; Felmer, Ricardo N

    2013-01-01

    Different culture systems have been studied that support development of somatic cell nuclear transfer (SCNT) embryos up to the blastocyst stage. However, the use of sequential and two-step culture systems has been less studied. The objective of the present study was to examine the developmental potential and quality of bovine SCNT embryos cultured in different two-step culture media based on KSOM, SOF and the macromolecules FBS and BSA (K-K/FBS, K-S/BSA and K-K/BSA, respectively). No differences were observed in the cleavage rate for any of the culture systems. However, there was a significant difference (Pculture system yielding a higher rate of blastocysts (28%) compared to other treatments (18 and 15%, for K-S/BSA and K-K/BSA, respectively). Although quality of embryos, as assessed by the total number of cells, was not different, the apoptosis index was significantly affected in the sequential culture system (K-S/BSA). Gene expression analysis showed alterations of DNMT1, IGF2, LIF, and PRDX6 genes in embryos cultured in K-S/FBS and of SOD2 in embryos cultured in K-K/BSA. In conclusion, we demonstrated that culture medium may affect not only the developmental potential of SCNT embryos but also, more importantly, the gene expression pattern and apoptotic index, presenting the possibility to manipulate the culture medium composition to modulate global gene expression and improve the overall efficiency of this technique.

  5. Solid-state fermentation of rice straw residues for its use as growing medium in ornamental nurseries

    Science.gov (United States)

    Belal, Elsayed B.; El-Mahrouk, M. E.

    2010-11-01

    This work was conducted at a private nursery in Kafr El-Sheikh governorate to investigate the bioconversion of rice straw into a soil-like substrate (SLS) by Phanerochaete chrysosporium and Trichoderma hazianum and the possibility of using rice straw compost in ornamental nurseries as a partial or total replacement of coconut peat (CP) and vermiculite (V) in the growing medium. The results showed that rice straw could be treated better by aerobic fermentation. The authors used five mixtures as follows: (1) Control (CP+V at 1:1 v/v), (2) SLS (100%), (3) SLS+CP (1:1 v/v), (4) SLS+V (1:1 v/v), and (5) SLS+CP+V (1:1:1 v/v/v). Data were recorded as seedling height, no. of leaves, shoot fresh and dry weights, root length and root fresh and dry weights in order to assess the quality of both transplants of Althea rosea (hollyhock) and Calendula officinalis (scotch marigold). Hollyhock seedlings grown in medium containing a mixture of SLS+CP+V displayed quality traits similar to those recorded from the control treatment, while scotch marigold seedlings in the same medium followed the control medium in quality.

  6. Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.

    Science.gov (United States)

    Kaiser, Andreas; Kale, Ajay; Novozhilova, Ekaterina; Siratirakun, Piyaporn; Aquino, Jorge B; Thonabulsombat, Charoensri; Ernfors, Patrik; Olivius, Petri

    2014-05-30

    Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Lippia dulcis shoot cultures as a source of the sweet sesquiterpene hernandulcin.

    Science.gov (United States)

    Sauerwein, M; Flores, H E; Yamazaki, T; Shimomura, K

    1991-04-01

    The axenic shoot culture of Lippia dulcis Trev., Verbenaceae, was established on hormone-free Murashige-Skoog solid medium containing 3% sucrose. Shoots were cultured in various liquid or solid media. Woody Plant liquid medium was best for shoot multiplication, but the production of hernandulcin was relatively low. The highest hernandulcin content (2.9% dry wt) was obtained after 28 days of culture on Murashige-Skoog solid medium containing 2% sucrose. The addition of chitosan to the culture media enhanced the growth of shoots as well as the production of hernandulcin, especially with the liquid medium.

  8. [Development and Evaluation of a New Selective Culture Medium, KBM Anaero RS-GNR, for Detection of Anaerobic Gram Negative Rods].

    Science.gov (United States)

    Narita, Taeko; Kato, Kyohei; Hanaiwa, Hiroki; Harada, Tetsuhiro; Funashima, Yumiko; Akiwa, Makoto; Sekiguchi, Jun-Ichiro; Nagasawa, Zenzo; Umemura, Tsukuru

    2017-03-22

    The laboratory culture methods for isolating drug-resistant pathogens has been the gold standard in medical microbiology, and play pivotal roles in the overall management of infectious diseases. Recently, several reports have emphasized the development of antibiotics-resistance among anaerobic gram-negative rods, especially Genus Bacteroides and Prevotella . Therefore, a selective culture method to detect these pathogens is needed. We developed here the new selective culture medium, termed "KBM Anaero RS-GNR," for detecting anaerobic Gram-negative rods. Growth capability and selectivity of the agar medium were assessed by using the pure culture suspensions of more than 100 bacterial strains as well as the 13 samples experimentally contaminated with these bacterial strains. This new medium, "KBM Anaero RS-GNR," successfully showed the selective isolation of anaerobic Gram-negative rods. Compared with commercially available medium, "PV Brucella HK Agar, " which is also designed to detect anaerobic Gram-negative rods, there was no significant difference of the overall detection efficiency between two media. However, "KBM Anaero RS-GNR" showed superior to selectivity for anaerobic Gram-negative rods, especially from the samples contaminated with Candida species. Thus, the culture method using KBM Anaero RS-GNR is relevant for isolation of anaerobic Gram-negative rods especially from clinical specimens.

  9. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture

    International Nuclear Information System (INIS)

    Yoshito, Daniele

    2011-01-01

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  10. Characterization of purified and Xerogel immobilized Novel Lignin Peroxidase produced from Trametes versicolor IBL-04 using solid state medium of Corncobs

    Directory of Open Access Journals (Sweden)

    Asgher Muhammad

    2012-08-01

    Full Text Available Abstract Background Cost-effective production of industrially important enzymes is a key for their successful exploitation on industrial scale. Keeping in view the extensive industrial applications of lignin peroxidase (LiP, this study was performed to purify and characterize the LiP from an indigenous strain of Trametes versicolor IBL-04. Xerogel matrix enzyme immobilization technique was applied to improve the kinetic and thermo-stability characteristics of LiP to fulfil the requirements of the modern enzyme consumer sector of biotechnology. Results A novel LiP was isolated from an indigenous T. versicolor IBL-04 strain. T. versicolor IBL-04 was cultured in solid state fermentation (SSF medium of corn cobs and maximum LiP activity of 592 ± 6 U/mL was recorded after five days of incubation under optimum culture conditions. The crude LiP was 3.3-fold purified with specific activity of 553 U/mg after passing through the DEAE-cellulose and Sephadex-G-100 chromatography columns. The purified LiP exhibited a relatively low molecular weight (30 kDa homogenous single band on native and SDS-PAGE. The LiP was immobilized by entrapping in xerogel matrix of trimethoxysilane (TMOS and proplytetramethoxysilane (PTMS and maximum immobilization efficiency of 88.6% was achieved. The free and immobilized LiPs were characterized and the results showed that the free and immobilized LiPs had optimum pH 6 and 5 while optimum temperatures were 60°C and 80°C, respectively. Immobilization was found to enhance the activity and thermo-stability potential of LiP significantly and immobilized LiP remained stable over broad pH and temperature range as compare to free enzyme. Kinetic constants Km and Vmax were 70 and 56 μM and 588 and 417 U/mg for the free and immobilized LiPs, respectively. Activity of this novel extra thermo-stable LiP was stimulated to variable extents by Cu2+, Mn2+ and Fe2+ whereas, Cystein, EDTA and Ag+ showed inhibitory effects

  11. A comparison of liquid and solid culture for determining relapse and durable cure in phase III TB trials for new regimens.

    Science.gov (United States)

    Phillips, Patrick P J; Mendel, Carl M; Nunn, Andrew J; McHugh, Timothy D; Crook, Angela M; Hunt, Robert; Bateson, Anna; Gillespie, Stephen H

    2017-11-24

    Tuberculosis kills more people than any other infectious disease, and new regimens are essential. The primary endpoint for confirmatory phase III trials for new regimens is a composite outcome that includes bacteriological treatment failure and relapse. Culture methodology is critical to the primary trial outcome. Patients in clinical trials can have positive cultures after treatment ends that may not necessarily indicate relapse, which was ascribed previously to laboratory cross-contamination or breakdown of old lesions. Löwenstein-Jensen (LJ) medium was the previous standard in clinical trials, but almost all current and future trials will use the Mycobacteria Growth Indicator Tube (MGIT) system due to its simplicity and consistency of use, which will affect phase III trial results. LJ was used for the definition of the primary endpoint in the REMoxTB trial, but every culture was also inoculated in parallel into the MGIT system. The data from this trial, therefore, provide a unique opportunity to investigate and compare the incidence of false 'isolated positives' in liquid and solid media and their potential impact on the primary efficacy results. All post-treatment positive cultures were reviewed in the REMoxTB clinical trial. Logistic regression models were used to model the incidence of isolated positive cultures on MGIT and LJ. A total of 12,209 sputum samples were available from 1652 patients; cultures were more often positive on MGIT than LJ. In 1322 patients with a favourable trial outcome, 126 (9.5%) had cultures that were positive in MGIT compared to 34 (2.6%) patients with positive cultures on LJ. Among patients with a favourable outcome, the incidence of isolated positives on MGIT differed by study laboratory (p cultures in some patients even after adjusting for laboratory, p cultures, positive MGIT cultures were more likely to be associated with higher grade TB symptoms reported within 7 days either side of sputum collection in patients with an

  12. Bioethanol production from sweet potato (Ipomoea batatas L. flour using co-culture of Trichoderma sp. and Saccharomyces cerevisiae in solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Manas Ranjan Swain

    2013-04-01

    Full Text Available The aim of this work was to study the optimiation of co-culturing of Trichoderma sp. and Saccharomyces cerevisiae (1:4 ratio on sweet potato (Ipomoea batatas L. flour (SPF for the production of bio-ethanol in solid-state fermentation (SSF. Maximum ethanol (172 g/kg substrate was produced in a medium containing 80% moisture, ammonium sulphate 0.2%, pH 5.0, inoculuted with 10% inoculum size and fermented at 30ºC for 72h. .Concomitant with highest ethanol concentration, maximum ethanol productivity (2.8 g/kg substrate/h, microbial biomass (23×10(8 CFU/ g substrate, ethanol yield (47 g/100g sugar consumed and fermentation efficiency (72% were also obtained under these conditions. Cell interaction was observed familiar between the viable cells of Trichoderma sp. and S. cerevisiae when co-cultured. Ethanol production ability by the co-culture was 65 % higher than the single culture of S. cerevisiae from un-saccharified SPF.

  13. Finite element method for viscoelastic medium with damage and the application to structural analysis of solid rocket motor grain

    Science.gov (United States)

    Deng, Bin; Shen, ZhiBin; Duan, JingBo; Tang, GuoJin

    2014-05-01

    This paper studies the damage-viscoelastic behavior of composite solid propellants of solid rocket motors (SRM). Based on viscoelastic theories and strain equivalent hypothesis in damage mechanics, a three-dimensional (3-D) nonlinear viscoelastic constitutive model incorporating with damage is developed. The resulting viscoelastic constitutive equations are numerically discretized by integration algorithm, and a stress-updating method is presented by solving nonlinear equations according to the Newton-Raphson method. A material subroutine of stress-updating is made up and embedded into commercial code of Abaqus. The material subroutine is validated through typical examples. Our results indicate that the finite element results are in good agreement with the analytical ones and have high accuracy, and the suggested method and designed subroutine are efficient and can be further applied to damage-coupling structural analysis of practical SRM grain.

  14. Computational study of plasma-solid interaction in DC glow discharge in argon plasma at medium pressures

    International Nuclear Information System (INIS)

    Havlickova, E; Bartos, P; Hrach, R

    2007-01-01

    In the presented contribution two groups of techniques of computational physics-fluid modelling and non self-consistent particle technique were used to study plasma-solid interaction in argon plasma. We focused both on the physical processes taking place in the sheath at various pressures and on the problems of computational physics. The attention was given to preparation of two-dimensional fluid models with realistic assumptions about physical processes taking place in plasma during the plasma-solid interaction, further to improvement of the non self-consistent technique of particle modelling, where the external electric field was obtained either from the fluid model or directly from the trajectories of charged particles and finally to efficiency of individual algorithms

  15. Selective versus non-selective culture medium for group B streptococcus detection in pregnancies complicated by preterm labor or preterm-premature rupture of membranes

    Directory of Open Access Journals (Sweden)

    Marcelo Luís Nomura

    Full Text Available The objective of this study was to identify group B streptococcus (GBS colonization rates and compare detection efficiency of selective versus non-selective culture media and anorectal versus vaginal cultures in women with preterm labor and preterm-premature rupture of membranes (PROM. A prospective cohort study of 203 women was performed. Two vaginal and two anorectal samples from each woman were collected using sterile swabs. Two swabs (one anorectal and one vaginal were placed separately in Stuart transport media and cultured in blood-agar plates for 48 hours; the other two swabs were inoculated separately in Todd-Hewitt selective media for 24 hours and then subcultured in blood-agar plates. Final GBS identification was made by the CAMP test. A hundred thrity-two cultures out of 812 were positive. The maternal colonization rate was 27.6%. Colonization rates were 30% for preterm PROM and 25.2% for preterm labor. Todd-Hewitt selective medium detected 87.5% and non-selective medium 60.7% GBS-positive women. Vaginal samples and anorectal samples had the same detection rate of 80.3%. Anorectal selective cultures detected 75% of carriers; 39% of GBS-positive women were detected only in selective medium. A combined vaginal-anorectal selective culture is appropriate for GBS screening in this population, minimizing laboratory costs.

  16. Hydrogen production characteristics of the organic fraction of municipal solid wastes by anaerobic mixed culture fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Li; Yu, Zhang [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China)]|[Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Zhenhong, Yuan; Yongming, Sun; Xiaoying, Kong [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China)

    2009-01-15

    The hydrogen production from the organic fraction of municipal solid waste (OFMSW) by anaerobic mixed culture fermentation was investigated using batch experiments at 37 C. Seven varieties of typical individual components of OFMSW including rice, potato, lettuce, lean meat, oil, fat and banyan leaves were selected to estimate the hydrogen production potential. Experimental results showed that the boiling treated anaerobic sludge was effective mixed inoculum for fermentative hydrogen production from OFMSW. Mechanism of fermentative hydrogen production indicates that, among the OFMSW, carbohydrates is the most optimal substrate for fermentative hydrogen production compared with proteins, lipids and lignocelluloses. This conclusion was also substantiated by experimental results of this study. The hydrogen production potentials of rice, potato and lettuce were 134 mL/g-VS, 106 mL/g-VS, and 50 mL/g-VS respectively. The hydrogen percentages of the total gas produced from rice, potato and lettuce were 57-70%, 41-55% and 37-67%. (author)

  17. Effects of exogenous growth regulators on cell suspension culture of yin-hong grape (vitis vinifera l.) and establishment of the optimum medium

    International Nuclear Information System (INIS)

    Chao, Y.; Feng, J.C.; Yan, W.Y.; Xiao, Y.; Jun, Y.Y

    2015-01-01

    Callus induced by stem of Yin-hong grape (Vitis vinifera L.) was used as materials and B5 medium as basic medium. The major growth parameters of cell suspension cultures with various levels of 1-Naphthaleneacetic acid (NAA) and 6-Benzyl aminopurine (6-BA) were investigated to provide a basis for the optimum medium of suspension cell cultures of Yin-hong grape regarding cell number, packed cell volume (PCV), dry cell weight (DCW), cell viability, and morphology. All data were analysed by of two-way analysis of variance (ANOVA). Results showed that the treatment of 6-BA and NAA would effect the cell growth dynamics, probably causing logarithmic phase in advance at higher levels of 6-BA. Different concentration of 6-BA and NAA had significant effects on cells number, PCV, DCW and viability (p<0.05), while no-significant effect was observed on the cells morphology. The optimum medium for suspension cell cultures of Yin-hong grape was identified as B5+1.5 mg/L6-BA+1.5 mg/LNAA+ 250 mg/L casein hydrolysate + 30 g/L sucrose. With the optimum medium, the maximum number of suspension cells after the logarithmic growth phase was 34.78 * 108 / mL, the highest cell viability reached 86.45%.; DCW reached 3.84 g/L and PCV reached 0.092 mL/mL after eight days cultivating. (author)

  18. Development of a versatile high-temperature short-time (HTST) pasteurization device for small-scale processing of cell culture medium formulations.

    Science.gov (United States)

    Floris, Patrick; Curtin, Sean; Kaisermayer, Christian; Lindeberg, Anna; Bones, Jonathan

    2018-07-01

    The compatibility of CHO cell culture medium formulations with all stages of the bioprocess must be evaluated through small-scale studies prior to scale-up for commercial manufacturing operations. Here, we describe the development of a bespoke small-scale device for assessing the compatibility of culture media with a widely implemented upstream viral clearance strategy, high-temperature short-time (HTST) treatment. The thermal stability of undefined medium formulations supplemented with soy hydrolysates was evaluated upon variations in critical HTST processing parameters, namely, holding times and temperatures. Prolonged holding times of 43 s at temperatures of 110 °C did not adversely impact medium quality while significant degradation was observed upon treatment at elevated temperatures (200 °C) for shorter time periods (11 s). The performance of the device was benchmarked against a commercially available mini-pilot HTST system upon treatment of identical formulations on both platforms. Processed medium samples were analyzed by untargeted LC-MS/MS for compositional profiling followed by chemometric evaluation, which confirmed the observed degradation effects caused by elevated holding temperatures but revealed comparable performance of our developed device with the commercial mini-pilot setup. The developed device can assist medium optimization activities by reducing volume requirements relative to commercially available mini-pilot instrumentation and by facilitating fast throughput evaluation of heat-induced effects on multiple medium lots.

  19. Hyperspectral imaging for presumptive identification of bacterial colonies on solid chromogenic culture media

    Science.gov (United States)

    Guillemot, Mathilde; Midahuen, Rony; Archeny, Delpine; Fulchiron, Corine; Montvernay, Regis; Perrin, Guillaume; Leroux, Denis F.

    2016-04-01

    BioMérieux is automating the microbiology laboratory in order to reduce cost (less manpower and consumables), to improve performance (increased sensitivity, machine algorithms) and to gain traceability through optimization of the clinical laboratory workflow. In this study, we evaluate the potential of Hyperspectral imaging (HSI) as a substitute to human visual observation when performing the task of microbiological culture interpretation. Microbial colonies from 19 strains subcategorized in 6 chromogenic classes were analyzed after a 24h-growth on a chromogenic culture medium (chromID® CPS Elite, bioMérieux, France). The HSI analysis was performed in the VNIR region (400-900 nm) using a linescan configuration. Using algorithms relying on Linear Spectral Unmixing, and using exclusively Diffuse Reflectance Spectra (DRS) as input data, we report interclass classification accuracies of 100% using a fully automatable approach and no use of morphological information. In order to eventually simplify the instrument, the performance of degraded DRS was also evaluated using only the most discriminant 14 spectral channels (a model for a multispectral approach) or 3 channels (model of a RGB image). The overall classification performance remains unchanged for our multispectral model but is degraded for the predicted RGB model, hints that a multispectral solution might bring the answer for an improved colony recognition.

  20. Application of Statistical Design to the Optimization of Culture Medium for Prodigiosin Production by Serratia marcescens SWML08

    Directory of Open Access Journals (Sweden)

    Venil, C. K.

    2009-01-01

    Full Text Available Combination of Plackett – Burman design (PBD and Box – Behnken design (BBD were applied for optimization of different factors for prodigiosin production by Serratia marcescens SWML08. Among 11 factors, incubation temperature, and supplement of (NH42PO4 and trace salts into the culture medium were selected due to significant positive effect on prodigiosin yield. Box - Behnken design, a response surface methodology, was used for further optimization of these selected factors for better prodigiosin output. Data were analyzed step wise and a second order polynomial model was established to identify the relationship between the prodigiosin output and the selected factors. The media formulations were optimized having the factors such as incubation temperature 30 °C, (NH42PO4 6 g/L and trace salts 0.6 g/L. The maximum experimental response for prodigiosin production was 1397.96 mg/L whereas the predicted value was 1394.26 mg/L. The high correlation between the predicted and observed values indicated the validity of the statistical design.

  1. Purification of Proteins From Cell-Culture Medium or Cell-Lysate by High-Speed Counter-Current Chromatography Using Cross-Axis Coil Planet Centrifuge

    Science.gov (United States)

    Shibusawa, Yoichi; Ito, Yoichiro

    2014-01-01

    This review describes protein purifications from cell culture medium or cell-lysate by high speed counter-current chromatography using the cross-axis coil planet centrifuge. Purifications were performed using aqueous two phase systems composed of polyethylene glycols and dextrans. PMID:25360182

  2. Enhancement of anti-candidal activity of endophytic fungus Phomopsis sp. ED2, isolated from Orthosiphon stamineus Benth, by incorporation of host plant extract in culture medium.

    Science.gov (United States)

    Yenn, Tong Woei; Lee, Chong Chai; Ibrahim, Darah; Zakaria, Latiffah

    2012-08-01

    This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anti-candidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.

  3. Optimization of cultural conditions for biosurfactant production by Pleurotus djamor in solid state fermentation.

    Science.gov (United States)

    Velioglu, Zulfiye; Ozturk Urek, Raziye

    2015-11-01

    Being eco-friendly, less toxic, more biodegradable and biocompatible, biological surfactants have higher activity and stability compared to synthetic ones. In spite of the fact that there are abundant benefits of biosurfactants over the synthetic congeners, the problem related with the economical and large scale production proceeds. The utilization of several industrial wastes in the production media as substrates reduces the production cost. This current study aims optimization of biosurfactant production conditions by Pleurotus djamor, grown on sunflower seed shell, grape wastes or potato peels as renewable cheap substrates in solid state fermentation. After determination of the best substrate for biosurfactant production, we indicate optimum size and amount of solid substrate, volume of medium, temperature, pH and Fe(2+) concentrations on biosurfactant production. In optimum conditions, by reducing water surface tension to 28.82 ± 0.3 mN/m and having oil displacement diameter of 3.9 ± 0.3 cm, 10.205 ± 0.5 g/l biosurfactant was produced. Moreover, chemical composition of biosurfactant produced in optimum condition was determined by FTIR. Lastly, laboratory's large-scale production was carried out in optimum conditions in a tray bioreactor designed by us and 8.9 ± 0.5 g/l biosurfactant was produced with a significant surface activity (37.74 ± 0.3 mN/m). With its economical suggestions and applicability of laboratory's large-scale production, this work indicates the possibility of using low cost agro-industrial wastes as renewable substrates for biosurfactant production. Therefore, using economically produced biosurfactant will reduce cost in several applications such as bioremediation, oil recovery and biodegradation of toxic chemicals. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  4. Quantitative proteome analysis of Streptomyces coelicolor Nonsporulating liquid cultures demonstrates a complex differentiation process comparable to that occurring in sporulating solid cultures

    DEFF Research Database (Denmark)

    Manteca, Angel; Jung, Hye R; Schwämmle, Veit

    2010-01-01

    involved in primary metabolism (ribosome, Krebs cycle, and energy production) were detected in greater abundance in MI. The most remarkable protein abundance differences between MII from solid and liquid cultures were associated with the final stages of hyphae compartmentalization and spore formation....

  5. The effect of ultrasonic pretreatment on biogas generation yield from organic fraction of municipal solid waste under medium solids concentration circumstance

    International Nuclear Information System (INIS)

    Rasapoor, Mazdak; Ajabshirchi, Yahya; Adl, Mehrdad; Abdi, Reza; Gharibi, Arash

    2016-01-01

    Highlights: • Ultrasonic pretreatment improved biogas yields at lower TS content samples. • Time of ultrasonication was significantly effective on maximum biogas yields. • Ultrasonic pretreatment can cause the release of TVFA content before digestion. • Specific energy between 5000 kJ/kg TS and 10,000 kJ/kg TS improved biogas yields. - Abstract: Hydrolysis is the most rate limiting step in almost all anaerobic digestion systems. To tackle long time duration, ultrasonic pretreatment proved that it can effectively improve biogas yield efficiency by effecting on soluble particles. In this study, the effect of three different ultrasonic power densities (0.2 W/mL, 0.4 W/mL and 0.6 W/mL) at three different times (10 min, 20 min and 30 min) on biogas yield of organic fraction of municipal solid waste (OFMSW) at three different total solid content (6%, 8% and 10%) were analyzed. Results showed significant (p < 0.01) effect of both sonication density and time of sonication on biogas final yield and biogas yield after 72 h digestion with the 6% TS content. Parameters like specific energy input and total volatile fatty acid (TVFA) content were also evaluated to find the best sonication treatments for OFMSW. For lower TS contents (6% and 8%), sonication treatment significantly (p < 0.01) increased TVFA concentration before digesting. It is also proved that specific energy input between 5000 kJ/kg TS and 10,000 kJ/kg TS can effectively increase the biogas yields, especially for 6% TS content, and caused maximum biogas yield produced after 72 h of digestion.

  6. Culture conditions and medium components for the production of mycelial biomass and exo-polysaccharides with Paecilomyces japonica in liquid culture.

    Science.gov (United States)

    Lee, Jong Seok; Jung, Woo Chul; Park, Seok Jae; Lee, Keun Eok; Shin, Won Cheol; Hong, Eock Kee

    2013-04-01

    In this study, the liquid culture conditions were optimized for maximal production of mycelial biomass and exo-polysaccharide by Paecilomyces japonica. The effects of medium composition, C/N ratio and physical parameters were investigated. From these experiments, 30 g glucose, 20 g yeast extract, 0.5 g KH2PO4, and 0.1 g CuCl2 2H2O in 1-l distilled water were found to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, agitation, and aeration were determined to be 27°C, uncontrolled pH, 400 rpm, and 1.0 vvm, respectively. Under these optimal conditions, the maximum mycelial growth and polysaccharides production were 23.1 g/l and 2.5 g/l, respectively. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  7. The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3

    International Nuclear Information System (INIS)

    Alper, O.; Yamaguchi, K.; Hitomi, J.; Honda, S.; Matsushima, T.; Abe, K.

    1990-01-01

    The Mr 185,000 glycoprotein encoded by human c-erbB-2/neu/HER2 gene, termed c-erbB-2 gene product, shows a close structural similarity with epidermal growth factor receptor and is now regarded to be a growth factor receptor for an as yet unidentified ligand. Abundant c-erbB-2 mRNA was demonstrated by Northern blot studies in the human breast cancer cell line SK-BR-3. Cellular radiolabeling experiments followed by immunoprecipitation with three different anti-c-erbB-2 gene product antibodies, recognizing extracellular domain, kinase domain, and carboxyl-terminal portion, respectively, demonstrated the production of a large amount of c-erbB-2 gene product which had the capacity to be phosphorylated. Immunization of mice with concentrated culture medium conditioned by SK-BR-3 cells always generated antibodies against c-erbB-2 gene product, demonstrating that this culture medium contained substance(s) immunologically indistinguishable from c-erbB-2 gene product. This observation was supported by the successful development of a monoclonal antibody against c-erbB-2 gene product, GFD-OA-p185-1, by immunizing mice with this culture medium. The biochemical nature of the substance(s) present in the culture medium was further characterized. When the culture medium conditioned by [35S]cysteine-labeled SK-BR-3 cells was immunoprecipitated by three different anti-c-erbB-2 gene product antibodies, only the antibody recognizing extracellular domain precipitated the [35S]-labeled protein with a molecular weight of 110,000, namely p110. The newly developed monoclonal antibody also immunoprecipitated this protein

  8. Economic and environmental review of Waste-to-Energy systems for municipal solid waste management in medium and small municipalities.

    Science.gov (United States)

    Fernández-González, J M; Grindlay, A L; Serrano-Bernardo, F; Rodríguez-Rojas, M I; Zamorano, M

    2017-09-01

    The application of Directive 2008/98/CE on Municipal Solid Waste (MSW) implies the need to introduce technologies to generate energy from waste. Incineration, the most widely used method, is difficult to implement in low populated areas because it requires a large amount of waste to be viable (100,000 tons per year). This paper analyses the economic and environmental costs of different MSW-to-Energy technologies (WtE) in an area comprising of 13 municipalities in southern Spain. We analyse anaerobic digestion (Biomethanization), the production of solid recovered fuel (SRF) and gasification, and compare these approaches to the present Biological Mechanical Treatment (BMT) with elimination of the reject in landfill, and incineration with energy recovery. From an economic standpoint the implementation of WtE systems reduces the cost of running present BMT systems and incineration; gasification presents the lowest value. From the environmental standpoint, Life Cycle Assessment shows that any WtE alternatives, including incineration, present important advantages for the environment when compared to BMT. Finally, in order to select the best alternative, a multi-criteria method is applied, showing that anaerobic digestion is the optimal solution for the area studied. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Temperature and pH optima of enzyme activities produced by cellulolytic thermophilic fungi in batch and solid-state cultures

    Energy Technology Data Exchange (ETDEWEB)

    Grajek, W

    1986-01-01

    The temperature and pH optima of cellulolytic activities produced by thermophilic fungi in liquid and solid-state cultures were established. Some differences in optimal conditions for enzyme activities, which depended on culture methods, were confirmed. 10 references.

  10. Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT embryos

    Directory of Open Access Journals (Sweden)

    María E Arias

    2013-01-01

    Full Text Available Different culture systems have been studied that support development of somatic cell nuclear transfer (SCNT embryos up to the blastocyst stage. However, the use of sequential and two-step culture systems has been less studied. The objective of the present study was to examine the developmental potential and quality of bovine SCNT embryos cultured in different two-step culture media based on KSOM, SOF and the macromolecules FBS and BSA (K-K/FBS, K-S/BSA and K-K/BSA, respectively. No differences were observed in the cleavage rate for any of the culture systems. However, there was a significant difference (P<0.01 in the rate of blastocyst development, with the K-K/ FBS culture system yielding a higher rate of blastocysts (28% compared to other treatments (18 and 15%, for K-S/BSA and K-K/BSA, respectively. Although quality of embryos, as assessed by the total number of cells, was not different, the apoptosis index was significantly affected in the sequential culture system (K-S/BSA. Gene expression analysis showed alterations of DNMT1, IGF2, LIF, and PRDX6 genes in embryos cultured in K-S/FBS and of SOD2 in embryos cultured in K-K/BSA. In conclusion, we demonstrated that culture medium may affect not only the developmental potential of SCNT embryos but also, more importantly, the gene expression pattern and apoptotic index, presenting the possibility to manipulate the culture medium composition to modulate global gene expression and improve the overall efficiency of this technique.

  11. Use of Gifu Anaerobic Medium for culturing 32 dominant species of human gut microbes and its evaluation based on short-chain fatty acids fermentation profiles.

    Science.gov (United States)

    Gotoh, Aina; Nara, Misaki; Sugiyama, Yuta; Sakanaka, Mikiyasu; Yachi, Hiroyuki; Kitakata, Aya; Nakagawa, Akira; Minami, Hiromichi; Okuda, Shujiro; Katoh, Toshihiko; Katayama, Takane; Kurihara, Shin

    2017-10-01

    Recently, a "human gut microbial gene catalogue," which ranks the dominance of microbe genus/species in human fecal samples, was published. Most of the bacteria ranked in the catalog are currently publicly available; however, the growth media recommended by the distributors vary among species, hampering physiological comparisons among the bacteria. To address this problem, we evaluated Gifu anaerobic medium (GAM) as a standard medium. Forty-four publicly available species of the top 56 species listed in the "human gut microbial gene catalogue" were cultured in GAM, and out of these, 32 (72%) were successfully cultured. Short-chain fatty acids from the bacterial culture supernatants were then quantified, and bacterial metabolic pathways were predicted based on in silico genomic sequence analysis. Our system provides a useful platform for assessing growth properties and analyzing metabolites of dominant human gut bacteria grown in GAM and supplemented with compounds of interest.

  12. Developing a Planting Medium from Solid Waste Compost and Construction and Demolition Rubble for Use in Quarry Rehabilitation

    Science.gov (United States)

    Assaf, E. A.

    2015-12-01

    The combination of construction, demolition and excavation (CDE) waste along with the increase in solid waste generation has put a major stress on Lebanon and on the management of its solid waste. Compounding this problem are the issues of quarries closure and rehabilitation and a decrease in forest and vegetative cover. This research aims to provide an integrated solution to the stated problem by developing a "soil mix" derived from a mélange of the organic matter of the solid waste (compost), the CDE waste, and soil. Excavation and construction debris were ground to several sizes and mixed with compost and soil at different ratios. Replicates of these mixes and a set of control (regular soil) were used. In this mix, native and indicator plants are planted (in pots). The plant species used are Mathiolla crassifolia and Zea mays (Corn). Results have shown successful growth of both corn and Mathiolla seedlings in the mixes with higher amounts of construction rubble and compost i.e. Rubble: Soil: Compost Ratio of 2:1:1 and 1:0:1. However treatments with no compost and with less quantities of rubble demonstrated the inability of the soil used to sustain plant growth alone (1:1:1 and 1:1:0). Last but not least, the control consisting of soil only ended up being the weakest mix with yellow corn leaves and small Mathiolla seedlings fifty days after planting and fertilizing. Additionally, soil analysis, rubble and compost analysis were conducted. The samples were tested for heavy metals, nutrient availability and values of pH and EC. No contamination has been reported and an abundance of macronutrients and micronutrients was documented for the soil and compost. High alkalinity is due to the presence of concrete and the high percentage of Calcium Carbonate in Lebanese soils. Accordingly, the most adequate mixes for planting are treatments A (2:1:1) and B (1:0:1) and they should be pursued for a pilot scale study to test their potential use in quarry rehabilitation and

  13. Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

    Science.gov (United States)

    Kaneko, Ai; Sankai, Yoshiyuki

    2014-01-01

    The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).

  14. Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

    Directory of Open Access Journals (Sweden)

    Ai Kaneko

    Full Text Available The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4 cells/mL (8.9×10(3 cells/cm2 without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm, greater cell viability (≥30% for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks.

  15. Enhancement in irradiated mononuclear cells in culture of mitogen-induced incorporation of [3H]thymidine by homologous conditioned medium

    International Nuclear Information System (INIS)

    Sandru, G.; Greiner, R.

    1994-01-01

    Incorporation of [ 3 H]thymidine in irradiated peripheral blood mononuclear cell cultures irradiated in vitro was stimulated significantly by either concanavalin A or phytohemagglutinin only in the presence of homologous conditioned medium. Production of this activity by mononuclear cells was enhanced by irradiation and/or pulsed exposure to puromycin but was abolished by actinomycin D. Addition of anti-interleukin 1 or anti-interleukin 2 monoclonal antibodies to the conditioned medium before assay did not influence the stimulatory action. A similar significant stimulation of mononuclear cell cultures irradiated with 6 Gy by concanavalin A was obtained when purified preparations of homologous conditioned medium were used in the assay. Purification was done by ultrafiltration and concentration, heparin agarose chromatography, ammonium sulfate precipitation, concanavalin A agarose chromatography, DEAE-ion exchange chromatography and HPLC gel filtration chromatography. With SDS-PAGE and silver staining, the active HPLC fraction gave one band of 50 kDa, suggesting that this protein is responsible for the co-stimulatory effect of homologous conditioned medium for both mitogen-induced irradiated and nonirradiated mononuclear cell cultures. 42 refs., 9 figs., 3 tabs

  16. Comparison of fermentation profiles between lupine and soybean by Aspergillus oryzae and Aspergillus sojae in solid-state culture systems

    NARCIS (Netherlands)

    Sardjono; Zhu, Y.; Knol, W.

    1998-01-01

    To explore the possibilities of using lupine as a soybean replacement in fermented foods, fermentation profiles of lupine and soybean by Aspergillus oryzae and A. sojae, respectively, in a solid-state culture were compared. Biomass, spore concentration, oxygen consumption rate, carbon dioxide

  17. Humanized medium (h7H) allows long-term primary follicular thyroid cultures from human normal thyroid, benign neoplasm, and cancer.

    Science.gov (United States)

    Bravo, Susana B; Garcia-Rendueles, Maria E R; Garcia-Rendueles, Angela R; Rodrigues, Joana S; Perez-Romero, Sihara; Garcia-Lavandeira, Montserrat; Suarez-Fariña, Maria; Barreiro, Francisco; Czarnocka, Barbara; Senra, Ana; Lareu, Maria V; Rodriguez-Garcia, Javier; Cameselle-Teijeiro, Jose; Alvarez, Clara V

    2013-06-01

    Mechanisms of thyroid physiology and cancer are principally studied in follicular cell lines. However, human thyroid cancer lines were found to be heavily contaminated by other sources, and only one supposedly normal-thyroid cell line, immortalized with SV40 antigen, is available. In primary culture, human follicular cultures lose their phenotype after passage. We hypothesized that the loss of the thyroid phenotype could be related to culture conditions in which human cells are grown in medium optimized for rodent culture, including hormones with marked differences in its affinity for the relevant rodent/human receptor. The objective of the study was to define conditions that allow the proliferation of primary human follicular thyrocytes for many passages without losing phenotype. Concentrations of hormones, transferrin, iodine, oligoelements, antioxidants, metabolites, and ethanol were adjusted within normal homeostatic human serum ranges. Single cultures were identified by short tandem repeats. Human-rodent interspecies contamination was assessed. We defined an humanized 7 homeostatic additives medium enabling growth of human thyroid cultures for more than 20 passages maintaining thyrocyte phenotype. Thyrocytes proliferated and were grouped as follicle-like structures; expressed Na+/I- symporter, pendrin, cytokeratins, thyroglobulin, and thyroperoxidase showed iodine-uptake and secreted thyroglobulin and free T3. Using these conditions, we generated a bank of thyroid tumors in culture from normal thyroids, Grave's hyperplasias, benign neoplasms (goiter, adenomas), and carcinomas. Using appropriate culture conditions is essential for phenotype maintenance in human thyrocytes. The bank of thyroid tumors in culture generated under humanized humanized 7 homeostatic additives culture conditions will provide a much-needed tool to compare similarly growing cells from normal vs pathological origins and thus to elucidate the molecular basis of thyroid disease.

  18. Antidepressant-like effects of a water-soluble extract from the culture medium of Ganoderma lucidum mycelia in rats.

    Science.gov (United States)

    Matsuzaki, Hirokazu; Shimizu, Yuta; Iwata, Naohiro; Kamiuchi, Shinya; Suzuki, Fumiko; Iizuka, Hiroshi; Hibino, Yasuhide; Okazaki, Mari

    2013-12-26

    Ganoderma lucidum is a popular medicinal mushroom used for promoting health and longevity in Asian countries. Previously, we reported that a water-soluble extract from a culture medium of Ganoderma lucidum mycelia (MAK) exerts antioxidative and cerebroprotective effects against ischemia-reperfusion injury in vivo. Here, we evaluated the antidepressant and anxiolytic activities of MAK in rats. MAK (0.3 or 1 g/kg, p.o.) was administered in the experimental animals 60 min before the forced swimming, open-field, elevated plus-maze, contextual fear-conditioning, and head twitch tests. Additionally, the mechanisms involved in the antidepressant-like action of MAK were investigated by the serotonin precursor 5-hydroxy-L-tryptophan (5-HTP)- or 5-HT2A agonist (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI)-induced head twitch responses. Treatment with MAK (1 g/kg) exhibited antidepressant-like effects in the forced swimming test, attenuated freezing behavior in the contextual fear-conditioning test, and decreased the number of head twitches induced by DOI, but not with 5-HTP. No significant response was observed in locomotion or anxiety-like behavior, when the animals were evaluated in the open-field or elevated plus-maze test, respectively. These data suggest that MAK has antidepressant-like potential, which is most likely due to the antagonism of 5-HT2A receptors, and possesses anxiolytic-like effects toward memory-dependent and/or stress-induced anxiety in rats.

  19. Application of a modified culture medium for the simultaneous counting of molds and yeasts and detection of aflatoxigenic strains of Aspergillus flavus and Aspergillus parasiticus.

    Science.gov (United States)

    Jaimez, J; Fente, C A; Franco, C M; Cepeda, A; Vázquez, B I

    2003-02-01

    Molds and yeasts from 91 samples of feed and raw materials used in feed formulation were enumerated on a new culture medium to which a beta cyclodextrin (beta-W7M 1.8-cyclodextrin) had been added. This medium was compared with other media normally used in laboratories for the routine analysis of fungi, such as Sabouraud agar, malt agar supplemented with 2% dextrose, and potato dextrose agar. When a t test for paired data (0.05 significance level, 95% confidence interval) was applied, no statistically significant differences between the results obtained with the new culture medium and those obtained with the other media used to enumerate molds and yeasts were found. For the evaluation of contamination due to aflatoxin for all of the samples, Sabouraud agar and yeast extract agar, both supplemented with 0.3% beta-W7M 1.8-cyclodextrin, and APA (aflatoxin-producing ability) medium were used. Aflatoxin was detected in 21% of the feed samples and in 23% of the raw-material samples analyzed, with maximal amounts of 2.8 and 6.0 microg of aflatoxin B1 per kg, respectively, being detected. In any case, the aflatoxin contents found exceeded the legally stipulated limits. The t test for paired data (0.05 significance level, 95% confidence interval) did not show statistically significant differences between the results obtained with the different culture media used for the detection of aflatoxins. The advantage of the new medium developed (Sabouraud agar with 0.3% beta-W7M 1.8-cyclodextrin) is that it allows simultaneous fungal enumeration and determination (under UV light) of the presence of aflatoxin-producing strains without prior isolation and culture procedures involving expensive and/or complex specific media and thus saves work, time, and money.

  20. CHROMagar COL-APSE: a selective bacterial culture medium for the isolation and differentiation of colistin-resistant Gram-negative pathogens

    DEFF Research Database (Denmark)

    Abdul Momin, Muhd Haziq F; Bean, David C; Hendriksen, Rene S.

    2017-01-01

    medium (SuperPolymyxin). Methodology. The medium was challenged with 84 isolates, including polymyxin B (POL B)-susceptible and -resistant type strains and colistin (COL)-resistant organisms recovered from human and animal samples. Susceptibility to COL and POL B was determined by agar dilution and broth...... microtitre dilution. The lower limit for the detection of COL-resistant organisms was also calculated for both CHROMagar COL-APSE and SuperPolymyxin media. The ability to isolate and correctly differentiate COL-resistant organisms within mixed cultures was also assessed and compared using both media. Results...

  1. Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

    Directory of Open Access Journals (Sweden)

    Holobiuc Irina

    2012-01-01

    Full Text Available Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.

  2. A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells.

    Science.gov (United States)

    Ahmadian Baghbaderani, Behnam; Tian, Xinghui; Scotty Cadet, Jean; Shah, Kevan; Walde, Amy; Tran, Huan; Kovarcik, Don Paul; Clarke, Diana; Fellner, Thomas

    2016-01-01

    Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.

  3. Effects of x-irradiation on cell division, oxygen consumption, and growth medium pH of an insect cell line cultured in vitro

    International Nuclear Information System (INIS)

    Koval, T.M.; Myser, W.C.; Hink, W.F.

    1975-01-01

    Cultured Trichoplusia ni cells in exponential growth were administered x-ray doses of 10,000 R and then subcultured. The untreated cell population began exponential growth within a few hours after subculture, eventually reaching stationary growth phase 96 hr later at a cell density of 2.08 x 10 6 cells/ml, whereas the irradiated cell population did not change for 24 hr after irradiation and then began exponential growth at a rate similar to that of control cells, also reaching stationary phase at 96 hr, but at a cell density of 0.93 x 10 6 cells/ml, which is less than half the maximum density of controls. From 24 to 96 hr after treatment, the x-irradiated cells were characterized by an increased consumption of oxygen that was nearly twice the amount utilized by control cells. The pH of the cell growth medium increases over 96 hr from 6.3 to 6.6 for irradiated as well as for untreated cultures, but since the number of x-rayed cells is less than half the number of untreated cells, the pH increase, per cell, of medium from irradiated cultures is about twice that of medium from control cultures

  4. Experimental investigation of the Rowe's dilatancy law on an atypical granular medium from a municipal solid waste incineration bottom ash

    Science.gov (United States)

    Becquart, Frédéric; Abriak, Nor Edine

    2013-06-01

    Municipal Solid Waste Incineration (MSWI) bottom ashes are irregular granular media because of their origin and are very heterogeneous with a large quantity of angular particles of different chemical species. MSWI bottom ash is a renewable granular resource alternative to the use of non-renewable standard granular materials. Beneficial use of these alternative granular materials mainly lies in road engineering. However, the studies about mechanical properties of such granular media still remain little developed, those being mainly based on empirical considerations. In this paper, a study of mechanical behaviour of a MSWI bottom ash under axisymmetric triaxial loadings conditions is presented. Samples are initially dense after Proctor compaction, are saturated and tested in drained conditions, under different effective confining pressures ranging from 100 to 600 kPa. The evolutions of volumetric strains show an initial contracting phase followed by a dilatancy phase, more pronounced when the confining pressure is low. The stresses ratios at the characteristic state and at the critical state appear in good agreement and with a null rate of volume variation. The angles of internal friction and dilatancy of the studied MSWI bottom ash are estimated and are similar to conventional granular materials used especially in road engineering. The dilatancy law of Rowe is well experimentally verified on this irregular recycled granular material.

  5. Effect of glycine and alanine supplementation on development of cattle embryos cultured in CR1aa medium with or without cumulus cells

    Directory of Open Access Journals (Sweden)

    Kr. BREDBACKA

    2008-12-01

    Full Text Available The effect of alanine (1 mM and glycine (10 mM supplementation on bovine embryo development in vitro was investigated. Presumptive bovine zygotes, produced by in vitro maturation and insemination of oocytes, were cultured for 144 h in CR1aa medium in the absence (Experiments 1 and 2 or presence of cumulus cells (Experiment 3. In Experiment 1, the proportion of morulae and blastocysts of cleaved embryos in glycine-supplemented medium was not different from that of the control medium (34% in both mediaglycine-enriched medium (69.5 vs. 53.3, P = 0.016. In Experiment 2, addition of alanine did not improve the formation of morulae and blastocysts (13% vs. 21% in control medium, and the mean cell numbers in morulae and blastocysts were lower than those in the control group (34.3 vs. 68.7, P = 0.007. In the presence of cumulus cells, the combined supplementation of glycine and alanine increased the proportion of morulae and blastocysts over that in the control medium (31% vs. 14%, P = 0.003.;

  6. One-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production using the whole solid-state fermentation medium of mixed filamentous fungi.

    Science.gov (United States)

    Maehara, Larissa; Pereira, Sandra C; Silva, Adilson J; Farinas, Cristiane S

    2018-02-01

    The efficient use of renewable lignocellulosic feedstocks to obtain biofuels and other bioproducts is a key requirement for a sustainable biobased economy. This requires novel and effective strategies to reduce the cost contribution of the cellulolytic enzymatic cocktails needed to convert the carbohydrates into simple sugars, in order to make large-scale commercial processes economically competitive. Here, we propose the use of the whole solid-state fermentation (SSF) medium of mixed filamentous fungi as an integrated one-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production. Ten different individual and mixed cultivations of commonly used industrial filamentous fungi (Aspergillus niger, Aspergillus oryzae, Trichoderma harzianum, and Trichoderma reesei) were performed under SSF and the whole media (without the extraction step) were used in the hydrolysis of pretreated sugarcane bagasse. The cocultivation of T. reesei with A. oryzae increased the amount of glucose released by around 50%, compared with individual cultivations. The release of glucose and reducing sugars achieved using the whole SSF medium was around 3-fold higher than obtained with the enzyme extract. The addition of soybean protein (0.5% w/w) during the hydrolysis reaction further significantly improved the saccharification performance by blocking the lignin and avoiding unproductive adsorption of enzymes. The results of the alcoholic fermentation validated the overall integrated process, with a volumetric ethanol productivity of 4.77 g/L.h, representing 83.5% of the theoretical yield. These findings demonstrate the feasibility of the proposed one-pot integrated strategy using the whole SSF medium of mixed filamentous fungi for on-site enzymes production, biomass hydrolysis, and ethanol production. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

  7. FRUITY AROMA PRODUCTION BY Ceratocystis fimbriata IN SOLID CULTURES FROM AGRO-INDUSTRIAL WASTES

    Directory of Open Access Journals (Sweden)

    Adriana Bramorski

    1998-09-01

    Full Text Available Solid state fermentations were carried out to test the efficacy of Ceratocystis fimbriata to grow on different agro-industrial substrates and aroma production. Seven media were prepared using cassava bagasse, apple pomace, amaranth and soya bean. All the media supported fungal growth. While amaranth medium produced pineapple aroma, media containing cassava bagasse, apple pomace and soya bean produced a strong fruity aroma. The aroma production was growth dependent and the maximum aroma intensity was detected a few hours before or after the maximum respirometric activity. Sixteen compounds were separated by gas cromatography of the components present in the headspace and fifteen of them were identified as acid (1, alcohols (6, aldehyde (1, ketones (2 and esters (5.Este estudo explorou a versatilidade de Ceratocystis fimbriata de crescer e produzir aromas naturais sobre substratos de resíduos agro-industriais. Bagaço de mandioca, bagaço de maçã, amaranto e soja em diferentes proporções compuseram os sete meios utilizados, mostrando ser substratos adequados para o crescimento e produção de aroma por este fungo em fermentação no estado sólido. Todos os meios contendo bagaço de mandioca, bagaço de maçã e soja em sua composição proporcionaram um forte aroma frutal, enquanto, o meio de amaranto produziu um agradável aroma de abacaxi. A produção de aroma foi dependente do crescimento, visto que a máxima intensidade do aroma foi detectado poucas horas antes ou depois da atividade respiratória máxima. Foram detectados dezesseis compostos pela cromatografia de gás no headspace das culturas, e quinze deles foram identificados: 1 ácido, 6 alcoois, 1 aldeído, 2 cetonas e 5 ésteres.

  8. Concept for a solid-state multi-parameter sensor system for cell-culture monitoring

    International Nuclear Information System (INIS)

    Baecker, M.; Beging, S.; Biselli, M.; Poghossian, A.; Wang, J.; Zang, W.; Wagner, P.; Schoening, M.J.

    2009-01-01

    In this study, a concept for a silicon-based modular solid-state sensor system for inline multi-parameter monitoring of cell-culture fermentation processes is presented. The envisaged multi-parameter sensor system consists of two identical sensor modules and is intended for continuous quantification of up to five (bio-)chemical and physical parameters, namely, glucose and glutamine concentration, pH value, electrolyte conductivity and temperature by applying different transducer principles and/or different operation modes. Experimental results for the field-effect electrolyte-insulator-semiconductor (EIS) sterilisable pH sensor and electrolyte conductivity sensor based on interdigitated electrodes are presented. The ongoing autoclaving does not have any significant impact on the pH-sensitive properties of a Ta 2 O 5 -gate EIS sensor. Even after 30 autoclaving cycles, the pH sensors show a clear pH response and nearly linear calibration curve with a slope of 57 ± 1 mV/pH. Additional scanning electron microscopy and ellipsometric investigations do not show any visible surface degradation or changes in the thickness of the pH-sensitive Ta 2 O 5 layer. The preliminary results demonstrate the suitability of the developed EIS sensor for an inline pH measurement during a fermentation process. In addition, interdigitated electrodes of different geometries serving as electrolyte conductivity sensor have been tested for measurements in relatively high ionic-strength solutions.

  9. Filamentous fungi and media for cellulase production in solid state cultures

    Science.gov (United States)

    Kilikian, B.V.; Afonso, L.C.; Souza, T.F.C.; Ferreira, R.G.; Pinheiro, I.R.

    2014-01-01

    Cellulase production was evaluated in two reference strains (T. reesei Rut-C30 and T. reesei QM9414), two strains isolated from a sugarcane cultivation area (Trichoderma sp. IPT778 and T. harzianum rifai IPT821) and one strain isolated in a program for biodiversity preservation in São Paulo state (Myceliophthora thermophila M77). Solid state cultures were performed using sugarcane bagasse (C), wheat bran (W) and/or soybean bran (S). The highest FPA was 10.6 U/gdm for M77 in SC (10:90) at 80% moisture, which was 4.4 times higher than production in pure W. C was a strong inducer of cellulase production, given that the production level of 6.1 U/gdm in WC (40:60) was 2.5 times higher than in pure W for strain M77; T. reesei Rut-C30 did not respond as strongly with about 1.6-fold surplus production. S advantageously replaced W, as the surplus production on SC (20:80) was 2.3 times relative to WC (20:80) for M77. PMID:24948946

  10. Aspergillus oryzae S2 alpha-amylase production under solid state fermentation: optimization of culture conditions.

    Science.gov (United States)

    Sahnoun, Mouna; Kriaa, Mouna; Elgharbi, Fatma; Ayadi, Dorra-Zouari; Bejar, Samir; Kammoun, Radhouane

    2015-04-01

    Aspergillus oryzae S2 was assayed for alpha-amylase production under solid state fermentation (SSF). In addition to AmyA and AmyB already produced in monitored submerged culture, the strain was noted to produce new AmyB oligomeric forms, in particular a dominant tetrameric form named AmyC. The latter was purified to homogeneity through fractional acetone precipitation and size exclusion chromatography. SDS-PAGE and native PAGE analyses revealed that, purified AmyC was an approximately 172 kDa tetramer of four 42 kDa subunits. AmyC was also noted to display the same NH2-terminal amino acid sequence residues and approximately the same physico-chemical properties of AmyA and AmyB, to exhibit maximum activity at pH 5.6 and 60 °C, and to produce maltose and maltotriose as major starch hydrolysis end-products. Soyabean meal was the best substitute to yeast extract compared to fish powder waste and wheat gluten waste. AmyC production was optimized under SSF using statistical design methodology. Moisture content of 76.25%, C/N substrate ratio of 0.62, and inoculum size of 10(6.87) spores allowed maximum activity of 22118.34 U/g of dried substrate, which was 33 times higher than the one obtained before the application of the central composite design (CCD). Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Rich Medium Composition Affects Escherichia coli Survival, Glycation, and Mutation Frequency during Long-Term Batch Culture.

    Science.gov (United States)

    Kram, Karin E; Finkel, Steven E

    2015-07-01

    Bacteria such as Escherichia coli are frequently grown to high density to produce biomolecules for study in the laboratory. To achieve this, cells can be incubated in extremely rich media that increase overall cell yield. In these various media, bacteria may have different metabolic profiles, leading to changes in the amounts of toxic metabolites produced. We have previously shown that stresses experienced during short-term growth can affect the survival of cells during the long-term stationary phase (LTSP). Here, we incubated cells in LB, 2× yeast extract-tryptone (YT), Terrific Broth, or Super Broth medium and monitored survival during the LTSP, as well as other reporters of genetic and physiological change. We observe differential cell yield and survival in all media studied. We propose that differences in long-term survival are the result of changes in the metabolism of components of the media that may lead to increased levels of protein and/or DNA damage. We also show that culture pH and levels of protein glycation, a covalent modification that causes protein damage, affect long-term survival. Further, we measured mutation frequency after overnight incubation and observed a correlation between high mutation frequencies at the end of the log phase and loss of viability after 4 days of LTSP incubation, indicating that mutation frequency is potentially predictive of long-term survival. Since glycation and mutation can be caused by oxidative stress, we measured expression of the oxyR oxidative stress regulator during log-phase growth and found that higher levels of oxyR expression during the log phase are consistent with high mutation frequency and lower cell density during the LTSP. Since these complex rich media are often used when producing large quantities of biomolecules in the laboratory, the observed increase in damage resulting in glycation or mutation may lead to production of a heterogeneous population of plasmids or proteins, which could affect the

  12. Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium.

    Science.gov (United States)

    Rojas-Martínez, Carmen; Rodríguez-Vivas, Roger Iván; Millán, Julio Vicente Figueroa; Bautista-Garfias, Carlos Ramón; Castañeda-Arriola, Roberto Omar; Lira-Amaya, José Juan; Urióstegui, Patricia Vargas; Carrasco, Juan José Ojeda; Martínez, Jesús Antonio Álvarez

    2018-04-01

    An attenuated live vaccine containing Babesia bovis and B. bigemina cultured in vitro with a serum-free medium was assessed for its clinical protection conferred of naïve cattle, under natural tick-challenge in a high endemicity zone to Babesia spp. Three groups of six animals were treated as follows: group I (GI) received a vaccine derived from parasites cultured with a free-serum medium; group II (GII) were immunized with the standard vaccine, with parasites cultured in a medium supplemented with 40% (v/v) bovine serum; and a control group (GIII) inoculated with non-infected bovine erythrocytes. Inocula were administered by IM route. Experimental animals were kept during 23days after vaccination in a cattle farm free of ticks and Babesia spp. Thereafter, cattle were moved to a high endemicity farm for natural exposure to Babesia spp. transmitted by Rhipicephalus microplus ticks. Protection against clinical babesiosis was observed in bovines belonging to GI (100%) and GII (83.33%), while the control animals (GIII) were not protected, and showed severe clinical signs, closely related to babesiosis, were observed for at least three consecutive days during the challenge. These were fever, anemia, which were measured simultaneously, and circulating parasites were detected by optic light microscopy. All cattle showed B. bovis and B. bigemina in stained blood films during the challenge; B. bovis antibody titers were higher than those to B. bigemina in GI and GII, and lower titers were determined in GIII. The protective capacity of the vaccine derived from B. bovis and B. bigemina cultured in vitro in a serum-free medium was demonstrated. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. New Culture Medium Containing Ionic Concentrations of Nutrients Similar to Concentrations Found in the Soil Solution †

    Science.gov (United States)

    Angle, J. Scott; McGrath, Stephen P.; Chaney, Rufus L.

    1991-01-01

    A new growth medium which closely approximates the composition of the soil solution is presented. This soil solution equivalent (SSE) medium contains the following components (millimolar): NO3, 2.5; NH4, 2.5; HPO4, 0.005; Na, 2.5; Ca, 4.0; Mg, 2.0; K, 0.503; Cl, 4.0; SO4, 5.0; ethylenediamine-di(o-hydroxyphenylacetic acid), 0.02; and MES [2-(N-morpholino)ethanesulfonic acid] (to maintain the pH at 6.0), 10, plus 0.1% arabinose. The advantages of the SSE medium are discussed. PMID:16348614

  14. Direct Succinic Acid Production from Minimally Pretreated Biomass Using Sequential Solid-State and Slurry Fermentation with Mixed Fungal Cultures

    Directory of Open Access Journals (Sweden)

    Jerico Alcantara

    2017-06-01

    Full Text Available Conventional bio-based succinic acid production involves anaerobic bacterial fermentation of pure sugars. This study explored a new route for directly producing succinic acid from minimally-pretreated lignocellulosic biomass via a consolidated bioprocessing technology employing a mixed lignocellulolytic and acidogenic fungal co-culture. The process involved a solid-state pre-fermentation stage followed by a two-phase slurry fermentation stage. During the solid-state pre-fermentation stage, Aspergillus niger and Trichoderma reesei were co-cultured in a nitrogen-rich substrate (e.g., soybean hull to induce cellulolytic enzyme activity. The ligninolytic fungus Phanerochaete chrysosporium was grown separately on carbon-rich birch wood chips to induce ligninolytic enzymes, rendering the biomass more susceptible to cellulase attack. The solid-state pre-cultures were then combined in a slurry fermentation culture to achieve simultaneous enzymatic cellulolysis and succinic acid production. This approach generated succinic acid at maximum titers of 32.43 g/L after 72 h of batch slurry fermentation (~10 g/L production, and 61.12 g/L after 36 h of addition of fresh birch wood chips at the onset of the slurry fermentation stage (~26 g/L production. Based on this result, this approach is a promising alternative to current bacterial succinic acid production due to its minimal substrate pretreatment requirements, which could reduce production costs.

  15. Isolation of a novel promoter for efficient protein expression by Aspergillus oryzae in solid-state culture.

    Science.gov (United States)

    Bando, Hiroki; Hisada, Hiromoto; Ishida, Hiroki; Hata, Yoji; Katakura, Yoshio; Kondo, Akihiko

    2011-11-01

    A novel promoter from a hemolysin-like protein encoding the gene, hlyA, was characterized for protein overexpression in Aspergillus oryzae grown in solid-state culture. Using endo-1,4-β-glucanase from A. oryzae (CelA) as the reporter, promoter activity was found to be higher than that of the α-amylase (amyA) and manganese superoxide dismutase (sodM) genes not only in wheat bran solid-state culture but also in liquid culture. Expression of the A. oryzae endoglucanase CelB and two heterologous endoglucanases (TrEglI and TrEglIII from Trichoderma reesei) under the control of the hlyA promoter were also found to be stronger than under the control of the amyA promoter in A. oryzae grown in wheat bran solid-state culture, suggesting that the hlyA promoter may be useful for the overproduction of other proteins as well. In wheat bran solid-state culture, the productivity of the hlyA promoter in terms of protein produced was high when the cultivation temperature was 30°C or 37°C, when the water content was 0.6 or 0.8 ml/g wheat bran, and from 48 to 72 h after inoculation. Because A. oryzae sporulated actively under these conditions and because hemolysin has been reported to play a role in fungal fruiting body formation, high-level expression of hlyA may be related to sporulation.

  16. Accurate and noninvasive embryos screening during in vitro fertilization (IVF) assisted by Raman analysis of embryos culture medium Accurate and noninvasive embryos screening during IVF

    Science.gov (United States)

    Shen, A. G.; Peng, J.; Zhao, Q. H.; Su, L.; Wang, X. H.; Hu, J. M.; Yang, J.

    2012-04-01

    In combination with morphological evaluation tests, we employ Raman spectroscopy to select higher potential reproductive embryos during in vitro fertilization (IVF) based on chemical composition of embryos culture medium. In this study, 57 Raman spectra are acquired from both higher and lower quality embryos culture medium (ECM) from 10 patients which have been preliminarily confirmed by clinical assay. Data are fit by using a linear combination model of least squares method in which 12 basis spectra represent the chemical features of ECM. The final fitting coefficients provide insight into the chemical compositions of culture medium samples and are subsequently used as criterion to evaluate the quality of embryos. The relative fitting coefficients ratios of sodium pyruvate/albumin and phenylalanine/albumin seem act as key roles in the embryo screening, attaining 85.7% accuracy in comparison with clinical pregnancy. The good results demonstrate that Raman spectroscopy therefore is an important candidate for an accurate and noninvasive screening of higher quality embryos, which potentially decrease the time-consuming clinical trials during IVF.

  17. Effects of zirconium and nitrogen plasma immersion ion implantation on the electrochemical corrosion behavior of Mg–Y–RE alloy in simulated body fluid and cell culture medium

    International Nuclear Information System (INIS)

    Jamesh, Mohammed Ibrahim; Wu, Guosong; Zhao, Ying; Jin, Weihong; McKenzie, David R.; Bilek, Marcela M.M.; Chu, Paul K.

    2014-01-01

    Highlights: • Dual Zr and N plasma ion implantation are conducted on WE43Mg alloy. • Zr and N implanted WE43 (ZrN-WE43) enhanced corrosion resistance in cell culture medium. • ZrN-WE43 enhanced corrosion resistance in simulated body fluid (SBF). • ZrN-WE43 shows near capacitive impedance spectra in cell culture medium. • Calcium phosphate is formed on the corrosion product. - Abstract: The effects of dual Zr and N plasma immersion ion implantation (PIII) on the corrosion behavior of WE43Mg alloy are evaluated in simulated body fluid (SBF) and cell culture medium (cDMEM). Zr and N PIII improves the corrosion resistance of WE43 which exhibits smaller i corr , larger R 1 and R 2 , smaller CPE 2 , and larger phase angle maxima in SBF and cDMEM. The Zr and N PIII WE43 samples exhibit 12-folds decrease in i corr in SBF and 71-folds decrease in i corr with near capacitive EIS in cDMEM. Analysis of the corrosion products reveals calcium phosphate

  18. Detection and characterization of silver nanoparticles and dissolved species of silver in culture medium and cells by AsFlFFF-UV-Vis-ICPMS: application to nanotoxicity tests.

    Science.gov (United States)

    Bolea, E; Jiménez-Lamana, J; Laborda, F; Abad-Álvaro, I; Bladé, C; Arola, L; Castillo, J R

    2014-03-07

    A methodology based on Asymmetric Flow Field-Flow Fractionation (AsFlFFF) coupled with UV-Vis absorption spectrometry and ICP mass spectrometry (ICPMS) has been developed and applied to the study of silver nanoparticles (AgNPs) and dissolved species of silver in culture media and cells used in cytotoxicity tests. The effect of a nano-silver based product (protein stabilized silver nanoparticles ca. 15 nm average diameter) on human hepatoma (HepG2) cell viability has been studied. UV-Vis absorption spectrometry provided information about the nature (organic vs. nanoparticle) of the eluted species, whereas the silver was monitored by ICPMS. A shift towards larger hydrodynamic diameters was observed in the AgNPs after a 24 hour incubation period in the culture medium, which suggests a "protein corona" effect. Silver(I) associated with proteins present in the culture medium has also been detected, as a consequence of the oxidation process experimented by the AgNPs. However, the Ag(I) released into the culture medium did not justify the toxicity levels observed. AgNPs associated with the cultured HepG2 cells were also identified by AsFlFFF, after applying a solubilisation process based on the use of tetramethylammonium hydroxide (TMAH) and Triton X-100. These results have been confirmed by transmission electronic microscopy (TEM) analysis of the fractions collected from the AsFlFFF. The effect of AgNPs on HepG2 cells has been compared to that caused by silver(I) as AgNO3 under the same conditions. The determination of the total content of silver in the cells confirms that a much larger mass of silver as AgNPs with respect to AgNO3 (16 to 1) is needed to observe a similar toxicity.

  19. Evaluation on the Effects of Culture Medium on Regeneration of Oil Palm Plantlets from Immature Embryos (IE)

    International Nuclear Information System (INIS)

    Wan Nur Syuhada, W.S.; Rasid, O.A.; Parveez, G.K.A

    2016-01-01

    Oil palm (Elaeis guineensis) tissue culture is slow, inefficient and genotype dependent. Therefore, efforts to improve the regeneration system are essential and thereby on-going. Immature embryos (IE) have been reported to be more responsive to in vitro culture as compared to other explant types. The objective of this study is to evaluate the most efficient culture media for the regeneration of whole plants from IE. IE were cultured on three media, namely N6, MY3 and MS +Y3. Initial observation showed that calli were induced as early as two weeks after culture. They started to produce friable calli around three to four months later. IE were shown to respond differently to the tested culture media. This initial result will be useful to identify the most responsive culture media for IE as explant. (author)

  20. Three-dimensional characterization of bacterial microcolonies on solid agar-based culture media.

    Science.gov (United States)

    Drazek, Laurent; Tournoud, Maud; Derepas, Frédéric; Guicherd, Maryse; Mahé, Pierre; Pinston, Frédéric; Veyrieras, Jean-Baptiste; Chatellier, Sonia

    2015-02-01

    For the last century, in vitro diagnostic process in microbiology has mainly relied on the growth of bacteria on the surface of a solid agar medium. Nevertheless, few studies focused in the past on the dynamics of microcolonies growth on agar surface before 8 to 10h of incubation. In this article, chromatic confocal microscopy has been applied to characterize the early development of a bacterial colony. This technology relies on a differential focusing depth of the white light. It allows one to fully measure the tridimensional shape of microcolonies more quickly than classical confocal microscopy but with the same spatial resolution. Placing the device in an incubator, the method was able to individually track colonies growing on an agar plate, and to follow the evolution of their surface or volume. Using an appropriate statistical modeling framework, for a given microorganism, the doubling time has been estimated for each individual colony, as well as its variability between colonies, both within and between agar plates. A proof of concept led on four bacterial strains of four distinct species demonstrated the feasibility and the interest of the approach. It showed in particular that doubling times derived from early tri-dimensional measurements on microcolonies differed from classical measurements in micro-dilutions based on optical diffusion. Such a precise characterization of the tri-dimensional shape of microcolonies in their late-lag to early-exponential phase could be beneficial in terms of in vitro diagnostics. Indeed, real-time monitoring of the biomass available in a colony could allow to run well established microbial identification workflows like, for instance, MALDI-TOF mass-spectrometry, as soon as a sufficient quantity of material is available, thereby reducing the time needed to provide a diagnostic. Moreover, as done for pre-identification of macro-colonies, morphological indicators such as three-dimensional growth profiles derived from

  1. A dedicated fungal culture medium is useful in the diagnosis of fungemia: a retrospective cross-sectional study.

    Science.gov (United States)

    Zheng, Shuwei; Ng, Tong Yong; Li, Huihua; Tan, Ai Ling; Tan, Thuan Tong; Tan, Ban Hock

    2016-01-01

    Mortality for candidemia ranges from 15% to 35%. Current guidelines recommend inoculating blood into three aerobic and three anaerobic blood culture bottles when candidemia is suspected, without mention of a fungal blood culture bottle. To determine the value of the BACTEC Myco/F Lytic blood culture media in the diagnosis of fungemia. A two-year retrospective cross-sectional study was performed for patients who had fungemia with submitted BACTEC Plus Aerobic/F (Aer), BACTEC Plus Anaerobic/F (Anaer) or Myco/F Lytic (Myco) blood culture bottles. The detection rate of fungemia was 77.4% in 93 patients with contemporaneously submitted blood culture bottles when limited to only Aer/Anaer culture results. The detection rate improved significantly with the addition of the Myco culture bottle results (pculture bottle submissions were less useful for patients with appropriate anti-fungal therapy administered within 48 hours [OR = 0.18, 95% CI = (0.06, 0.49), p = 0.001] and those with fungal growth detected within 48 hours [OR = 0.33, 95% CI = (0.12, 0.89), p = 0.001]. Among a subset of patients with concordant blood culture results, those with Myco culture bottles submission allowed earlier fungal detection and speciation by at least one day in 27.5% and 25.0% of the cases respectively. Our study highlights the importance of a dedicated fungal blood culture when fungemia is clinically suspected. Nearly a quarter of fungemias may be missed if a fungal blood culture is not performed.

  2. Optimization of a serum-free culture medium for mouse embryonic stem cells using design of experiments (DoE) methodology.

    Science.gov (United States)

    Knöspel, Fanny; Schindler, Rudolf K; Lübberstedt, Marc; Petzolt, Stephanie; Gerlach, Jörg C; Zeilinger, Katrin

    2010-12-01

    The in vitro culture behaviour of embryonic stem cells (ESC) is strongly influenced by the culture conditions. Current culture media for expansion of ESC contain some undefined substances. Considering potential clinical translation work with such cells, the use of defined media is desirable. We have used Design of Experiments (DoE) methods to investigate the composition of a serum-free chemically defined culture medium for expansion of mouse embryonic stem cells (mESC). Factor screening analysis according to Plackett-Burman revealed that insulin and leukaemia inhibitory factor (LIF) had a significant positive influence on the proliferation activity of the cells, while zinc and L: -cysteine reduced the cell growth. Further analysis using minimum run resolution IV (MinRes IV) design indicates that following factor adjustment LIF becomes the main factor for the survival and proliferation of mESC. In conclusion, DoE screening assays are applicable to develop and to refine culture media for stem cells and could also be employed to optimize culture media for human embryonic stem cells (hESC).

  3. The population of phosphate solubilizing bacteria (PSB from Cikaniki, Botol Mountain, and Ciptarasa Area, and the ability of PSB to solubilize insoluble P in solid pikovskaya medium

    Directory of Open Access Journals (Sweden)

    SULIASIH

    2006-04-01

    Full Text Available Phosphate solubilizing bacteria (PSB were collected from plant rhizosphere at Cikaniki (1100 m asl., soil at Botol Mountain (1000, 1500, and 1800 m asl., and Ciptarasa (600, 1000, and 1500 m asl., area at Gunung Halimun National Park (GHNP. The soil ware collected randomly from 3 areas and taken from 0-15 cm depth in the plants rhizosphere at Cikaniki and forest floor soil in Gunung Botol and Ciptarasa. The result showed that the difference of elevation area, soil pH, forest vegetation, and microbial habitat (rhizosphere and forest floor were not the inhabitation factors of the biodiversity of PSB and their ability to solubilize insoluble phosphate, but the inhabitation factors on the growth of the PSB population. The highest population of PSB at GHNP was founded in the plant rhizosphere of Altingia exelsa Norona and Schima wallichii (Dc. Korth (107sel/g soil at Cikaniki and in the forest floor soil (108sel/g soil at Botol Mountain (1000 m asl.. Pseudomonas sp., Bacillus sp., Bacillus megaterium, and Chromobacterium sp. dominated Cikaniki, Botol Mountain and Ciptarasa area. Those isolates could solubilize insoluble phosphate on solid Pikovskaya medium with the range of diameter is 1.5-2.5 cm.

  4. Sporothrix schenckii Sensu Lato identification in fragments of skin lesion cultured in NNN medium for differential diagnosis of cutaneous leishmaniasis.

    Science.gov (United States)

    Antonio, Liliane de Fátima; Pimentel, Maria Inês Fernandes; Lyra, Marcelo Rosandiski; Madeira, Maria de Fátima; Miranda, Luciana de Freitas Campos; Paes, Rodrigo Almeida; Brito-Santos, Fábio; Carvalho, Maria Helena Galdino Figueredo; Schubach, Armando de Oliveira

    2017-02-01

    Eighty-nine patients with clinical suspicion of leishmaniasis were referred for differential diagnosis. Sporothrix schenckii sensu lato was isolated in Novy-MacNeal-Nicolle + Schneider media in 98% of 64 patients with final diagnosis of sporotrichosis. This medium may be suitable for diagnosis of sporotrichosis in areas where cutaneous leishmaniasis is also endemic. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Efficient mannitol production by wild-type Lactobacillus reuteri CRL 1101 is attained at constant pH using a simplified culture medium.

    Science.gov (United States)

    Ortiz, Maria Eugenia; Raya, Raúl R; Mozzi, Fernanda

    2015-10-01

    Mannitol is a natural polyol with multiple industrial applications. In this work, mannitol production by Lactobacillus reuteri CRL 1101 was studied at free- and controlled-pH (6.0-4.8) fermentations using a simplified culture medium containing yeast and beef extracts and sugarcane molasses. The activity of mannitol 2-dehydrogenase (MDH), the enzyme responsible for mannitol synthesis, was determined. The effect of the initial biomass concentration was further studied. Mannitol production (41.5 ± 1.1 g/l), volumetric productivity (Q Mtl 1.73 ± 0.05 g/l h), and yield (Y Mtl 105 ± 11 %) were maximum at pH 5.0 after 24 h while the highest MDH activity (1.66 ± 0.09 U/mg protein) was obtained at pH 6.0. No correlation between mannitol production and MDH activity was observed when varying the culture pH. The increase (up to 2000-fold) in the initial biomass concentration did not improve mannitol formation after 24 h although a 2-fold higher amount was produced at 8 h using 1 or 2 g cell dry weight/l comparing to the control (0.001 g cell dry weight/l). Finally, mannitol isolation under optimum fermentation conditions was achieved. The mannitol production obtained in this study is the highest reported so far by a wild-type L. reuteri strain and, more interestingly, using a simplified culture medium.

  6. Effect of Zn and Mg in tricalcium phosphate and in culture medium on apoptosis and actin ring formation of mature osteoclasts

    International Nuclear Information System (INIS)

    Li Xia; Ito, Atsuo; Sogo, Yu; Senda, Koji; Yamazaki, Atsushi

    2008-01-01

    This study investigated the resorptive activity of osteoclasts on tricalcium phosphate (TCP), zinc-containing tricalcium phosphate (ZnTCP) and magnesium-containing tricalcium phosphate (MgTCP) ceramics in different Zn- or Mg-containing culture media. On the TCP ceramic, an increase in Zn ions in the culture medium within the range between 0.3 and 6.8 ppm significantly induced an increase in osteoclast apoptosis and a decrease in actin ring formation. However, even a high level of Mg ions up to 100 ppm in the culture medium was unlikely to induce an increase in osteoclast apoptosis. Mg ions in the MgTCP ceramics have no effect on osteoclast apoptosis and actin ring formation. There was almost no significant difference in osteoclast apoptosis and actin ring formation between ZnTCP and MgTCP ceramics which have the same solubility and dissolution rates. It is indicated that only an increase in Zn level outside resorption lacuna has an inhibitory effect on osteoclast resorption and that an increase in Zn level inside resorption lacuna could not influence the osteoclast activity.

  7. Medium selection for exopolysaccharide and biomass production in submerged cultures of culinary-medicinal mushrooms from Turkey

    NARCIS (Netherlands)

    Kizilcik, M.; Yamaç, M.; Griensven, van L.J.L.D.

    2010-01-01

    The present study investigates the exopolysaccharide (EPS) and biomass production of 18 strains of 15 species of culinary-medicinal higher Basidiomycetes in submerged culture under four different media. Gloeophyllum abietinum and Schizophyllum commune produced the highest EPS and biomass

  8. Defined Essential 8™ Medium and Vitronectin Efficiently Support Scalable Xeno-Free Expansion of Human Induced Pluripotent Stem Cells in Stirred Microcarrier Culture Systems

    Science.gov (United States)

    Badenes, Sara M.; Fernandes, Tiago G.; Cordeiro, Cláudia S. M.; Boucher, Shayne; Kuninger, David; Vemuri, Mohan C.; Diogo, Maria Margarida; Cabral, Joaquim M. S.

    2016-01-01

    Human induced pluripotent stem (hiPS) cell culture using Essential 8™ xeno-free medium and the defined xeno-free matrix vitronectin was successfully implemented under adherent conditions. This matrix was able to support hiPS cell expansion either in coated plates or on polystyrene-coated microcarriers, while maintaining hiPS cell functionality and pluripotency. Importantly, scale-up of the microcarrier-based system was accomplished using a 50 mL spinner flask, under dynamic conditions. A three-level factorial design experiment was performed to identify optimal conditions in terms of a) initial cell density b) agitation speed, and c) to maximize cell yield in spinner flask cultures. A maximum cell yield of 3.5 is achieved by inoculating 55,000 cells/cm2 of microcarrier surface area and using 44 rpm, which generates a cell density of 1.4x106 cells/mL after 10 days of culture. After dynamic culture, hiPS cells maintained their typical morphology upon re-plating, exhibited pluripotency-associated marker expression as well as tri-lineage differentiation capability, which was verified by inducing their spontaneous differentiation through embryoid body formation, and subsequent downstream differentiation to specific lineages such as neural and cardiac fates was successfully accomplished. In conclusion, a scalable, robust and cost-effective xeno-free culture system was successfully developed and implemented for the scale-up production of hiPS cells. PMID:26999816

  9. Bacillus coagulans tolerance to 1-ethyl-3-methylimidazolium-based ionic liquids in aqueous and solid-state thermophilic culture.

    Science.gov (United States)

    Simmons, Christopher W; Reddy, Amitha P; Vandergheynst, Jean S; Simmons, Blake A; Singer, Steven W

    2014-01-01

    The use of ionic liquids (ILs) to disrupt the recalcitrant structure of lignocellulose and make polysaccharides accessible to hydrolytic enzymes is an emerging technology for biomass pretreatment in lignocellulosic biofuel production. Despite efforts to reclaim and recycle IL from pretreated biomass, residual IL can be inhibitory to microorganisms used for downstream fermentation. As a result, pathways for IL tolerance are needed to improve the activity of fermentative organisms in the presence of IL. In this study, microbial communities from compost were cultured under high-solids and thermophilic conditions in the presence of 1-ethyl-3-methylimidazolium-based ILs to enrich for IL-tolerant microorganisms. A strain of Bacillus coagulans isolated from an IL-tolerant community was grown in liquid and solid-state culture in the presence of the ILs 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]) or 1-ethyl-3-methylimidazolium chloride ([C2mim][Cl]) to gauge IL tolerance. Viability and respiration varied with the concentration of IL applied and the type of IL used. B. coagulans maintained growth and respiration in the presence of 4 wt% IL, a concentration similar to that present on IL-pretreated biomass. In the presence of both [C2mim][OAc] and [C2mim][Cl] in liquid culture, B. coagulans grew at a rate approximately half that observed in the absence of IL. However, in solid-state culture, the bacteria were significantly more tolerant to [C2mim][Cl] compared with [C2mim][OAc]. B. coagulans tolerance to IL under industrially relevant conditions makes it a promising bacterium for understanding mechanisms of IL tolerance and discovering IL tolerance pathways for use in other microorganisms, particularly those used in bioconversion of IL-pretreated plant biomass. © 2013 American Institute of Chemical Engineers.

  10. Release of somatic embryogenic potential from excised zygotic embryos of carrot and maintenance of proembryonic cultures in hormone-free medium

    Science.gov (United States)

    Smith, D. L.; Krikorian, A. D.

    1989-01-01

    Excised zygotic embryos, mericarps ("seeds") and hypocotyls of seedlings of cultivated carrot Daucus carota cv. Scarlet Nantes were evaluated for their ability to generate somatic embryos on a semisolid hormone-free nutrient medium. Neither intact zygotic embryos nor hypocotyls ever produced somatic embryos. However, mericarps and broken zygotic embryos were excellent sources for somatic embryo production (response levels as high as 86%). Somatic embryo formation was highest from cotyledons, but was also observed on isolated hypocotyls and root tips of mature zygotic embryos. On media containing unreduced nitrogen, somatic embryo formation led to the generation of vigorous cultures comprised entirely of somatic embryos at various stages of development which in turn proliferated still other somatic embryos. However, a medium was devised which when 1-5 mM NH4+ was the sole nitrogen source, led only to a proliferation of globular proembryos. Sustained subculturing of these proembryos at 2-3 week intervals enabled establishment of highly uniform cultures in which no further development into more mature stages of embryonic development occurred. These have been maintained, without decline, as morphogenetically competent proembryonic globules for over ten months. A basal medium containing from 1-5 mM NH4+ as the sole nitrogen source appears not to be inductive to somatic proembryo formation. Instead, such a medium is best thought of as permissive to the expression of embryogenically determined cells within zygotic embryos. By excising and breaking or wounding zygotic embryos, constituent cells are probably released from positional or chemical restraints and thus are able to express their innate embryogenic potential. Once a proembryonic culture is established, this medium containing 1-5 mM NH4+ as the sole nitrogen source provides a nonpermissive environment to the development and growth of later embryonic stages, but it does allow the continued formation and

  11. The amorphous solid dispersion of the poorly soluble ABT-102 forms nano/microparticulate structures in aqueous medium: impact on solubility

    Directory of Open Access Journals (Sweden)

    Frank KJ

    2012-11-01

    Full Text Available Kerstin J Frank,1,3 Ulrich Westedt,2 Karin M Rosenblatt,2 Peter Hölig,2 Jörg Rosenberg,2 Markus Mägerlein,2 Gert Fricker,3 Martin Brandl11Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense, Denmark; 2Abbott GmbH and Co. KG, Ludwigshafen, Germany; 3Department of Pharmaceutical Technology, University of Heidelberg, Heidelberg, GermanyAbstract: Amorphous solid dispersions (ASDs are a promising formulation approach for poorly soluble active pharmaceutical ingredients (APIs, because they ideally enhance both dissolution rate and solubility. However, the mechanism behind this is not understood in detail. In the present study, we investigated the supramolecular and the nano/microparticulate structures that emerge spontaneously upon dispersion of an ASD in aqueous medium and elucidated their influence on solubility. The ASD, prepared by hot melt extrusion, contained the poorly soluble ABT-102 (solubility in buffer, 0.05 µg/mL, a hydrophilic polymer, and three surfactants. The apparent solubility of ABT-102 from the ASD-formulation was enhanced up to 200 times in comparison to crystalline ABT-102. At the same time, the molecular solubility, as assessed by inverse equilibrium dialysis, was enhanced two times. Asymmetrical flow field-flow fractionation in combination with a multiangle light-scattering detector, an ultraviolet detector, and a refractometer enabled us to separate and identify the various supramolecular assemblies that were present in the aqueous dispersions of the API-free ASD (placebo and of binary/ternary blends of the ingredients. Thus, the supramolecular assemblies with a molar mass between 20,000 and 90,000 could be assigned to the polyvinylpyrrolidone/vinyl acetate 64, while two other kinds of assemblies were assigned to different surfactant assemblies (micelles. The amount of ABT-102 remaining associated with each of the assemblies upon fractionation was quantified offline with high

  12. Induced Biosynthesis of resveratrol and the prenylated stilbenoids arachidan-1 and arachidan-3 in hairy root cultures of peanut: effects of culture medium and growth stage

    Science.gov (United States)

    The peanut plant has evolved specialized biosynthetic mechanisms that allowed resisting infection by producing diverse secondary metabolites. Among these unique compounds are the stilbenoids, which include resveratrol analogues. Our previous research demonstrated that peanut hairy root cultures prov...

  13. Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.

    Science.gov (United States)

    Pisciolaro, Ricardo Luiz; Duailibi, Monica Talarico; Novo, Neil Ferreira; Juliano, Yara; Pallos, Debora; Yelick, Pamela Crotty; Vacanti, Joseph Phillip; Ferreira, Lydia Masako; Duailibi, Silvio Eduardo

    2015-11-01

    One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I (p = 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV (p = 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.

  14. Detection of microorganisms in culture medium through the neutron radiographic technique; Deteccao de microorganismos em meios de cultura pela tecnica de neutrongrafia

    Energy Technology Data Exchange (ETDEWEB)

    Wacha, Reinaldo

    1999-05-01

    The study aims to obtain a more effective and faster method for the detection of bacteria in several culture media, such as potable water and blood. After the process growth in the culture medium, separation and suspension in buffer solution based in boron, the bacteria are deposited in track detectors that are submitted to thermal neutron beams (neutron flux: 2,2 x 10{sup 5} n.cm{sup -2}.s{sup -1}), resulting from the channel J-9 of the Argonauta research reactor, from the Nuclear Engineering Institute, IEN/CNEN. The latent tracks arisen from the alpha particles proceeding from the reaction B ({eta}, {alpha}) Li and, after having been revealed, are analyzed by an optical microscope that allows to detect the existence of the bacteria. Afterwards, they were analyzed in a nanoscope which helps the identification of the tracks of the alpha particles. (author)

  15. A salting out system for improving the efficiency of the headspace solid-phase microextraction of short and medium chain free fatty acids.

    Science.gov (United States)

    Fiorini, Dennis; Pacetti, Deborah; Gabbianelli, Rosita; Gabrielli, Serena; Ballini, Roberto

    2015-08-28

    Given the importance of short and medium chain free fatty acids (FFAs) in several fields, this study sought to improve the extraction efficiency of the solid-phase microextraction (SPME) of FFAs by evaluating salting out agents that appear promising for this application. The salts ammonium sulfate ((NH4)2SO4) and sodium dihydrogen phosphate (NaH2PO4) were tried on their own and in combination (3.7/1), in four different total amounts, as salting out agents in the headspace-SPME-gas chromatographic (HS-SPME-GC) analysis of the FFAs from acetic acid (C2) to decanoic acid (C10). Their performance in a model system of an aqueous standard mixture of FFAs at a pH of 3.5 was compared to that of the more commonly used sodium chloride (NaCl) and sodium sulfate (Na2SO4). All of the salts and salt systems evaluated, in proper amount, gave improved results compared to NaCl (saturated), which instead gave interesting results only for the least volatile FFAs C8 and C10. For C2-C6, the salt system that gave the best results compared to NaCl was (NH4)2SO4/NaH2PO4, in the highest of the four amounts evaluated, with factor increases between 1.2 and 4.1-fold, and NaH2PO4, between 1.0 and 4.3-fold. The SPME extraction efficiency given by the mixture (NH4)2SO4/NaH2PO4 was also assessed on biological and food samples, confirming that overall it performed better than NaCl. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Could Externalized St. Jude Medical Riata® Lead Be a Culture Medium of a Polymicrobial Endocarditis? A Clinical Case

    Directory of Open Access Journals (Sweden)

    Zefferino Palamà

    2017-01-01

    Full Text Available We report the case of a man affected by polymicrobial endocarditis developed on a St. Jude Medical Riata lead with a malfunction because of the outsourcing of conductors. The patient was treated with antibiotic targeted therapy and showed different bacteria at the blood cultures and then underwent transvenous leads extraction. Vegetations were highlighted on the caval, atrial, and ventricular tracts of the Riata lead, but the cultures were all negative. The externalization of Riata lead may cause the malfunction but it could also promote bacterial colonies and vegetations. In conclusion, looking for early signs of infection is mandatory during Riata leads follow-up checks.

  17. Utilizing soybean milk to culture soybean pathogens

    Science.gov (United States)

    Liquid and semi-solid culture media are used to maintain and proliferate bacteria, fungi, and Oomycetes for research in microbiology and plant pathology. In this study, a comparison was made between soybean milk medium, also referred to as soymilk, and media traditionally used for culturing soybean ...

  18. The Cultural Orientations of Entrepreneurs and Employees' Job Satisfaction: The Turkish Small and Medium Sized Enterprises (SMEs) Case

    Science.gov (United States)

    Yetim, Nalan; Yetim, Unsal

    2006-01-01

    The primary purpose of this study was to determine whether cultural orientations which were pervasive and salient in the society of SMEs' entrepreneurs predict employees' job satisfaction. Paternalism, collectivism, individualism, power distance, uncertainty avoidance were assessed as pervasive and salient attributes for Turkish society. Data were…

  19. Surface characterization of insulin-coated Ti6Al4V medical implants conditioned in cell culture medium: An XPS study

    Energy Technology Data Exchange (ETDEWEB)

    Shchukarev, Andrey, E-mail: andrey.shchukarev@umu.se [Department of Chemistry, Umeå University, Umeå SE-90187 (Sweden); Malekzadeh, Behnosh Öhrnell [Department of Orthodontics, Institute of Odontology, Sahlgrenska Academy, University of Gothenburg, SE-40530 (Sweden); Department of Biomaterials, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, SE-40530 (Sweden); Ransjö, Maria [Department of Orthodontics, Institute of Odontology, Sahlgrenska Academy, University of Gothenburg, SE-40530 (Sweden); Tengvall, Pentti [Department of Biomaterials, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, SE-40530 (Sweden); Westerlund, Anna [Department of Orthodontics, Institute of Odontology, Sahlgrenska Academy, University of Gothenburg, SE-40530 (Sweden)

    2017-04-15

    Highlights: • In the absence of FBS, chemically immobilized insulin layer remains intact; • The immobilized insulin expose hydrophobic domains outward the implant; • In the presence of FBS, a partial replacement of insulin occurs; • The immobilized insulin stabilizes the secondary structure of adsorbed proteins. - Abstract: Surface characterization of insulin-coated Ti6Al4V medical implants, after incubation in α-minimum essential medium (α-MEM), was done by X-ray photoelectron spectroscopy (XPS), in order to analyze the insulin behavior at the implant – α-MEM interface. In the absence of serum proteins in cell culture medium, the coated insulin layer remained intact, but experienced a time-dependent structural transformation exposing hydrophobic parts of the protein toward the solution. The presence of fetal bovine serum (FBS) in the medium resulted in partial substitution of insulin by serum proteins. In spite of some insulin release, the remaining coated layer demonstrated a direct surface effect by stabilizing the structure of protein competitors, and by supporting the accumulation of calcium and phosphate ions at the interface. A structurally stable protein layer with incorporated calcium and phosphate ions at the implant–tissue interface could be an important prerequisite for enhanced bone formation.

  20. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

    Science.gov (United States)

    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Optimum conditions for growth in liquid medium of Oscillatoria formosa Bory used as the principal food in laboratory culture of intermediate hosts for schistosomosis and fasciolosis

    Directory of Open Access Journals (Sweden)

    Ferreira Filipa M.

    2000-09-01

    Full Text Available The rearing of snails, intermediate hosts of Schistosoma haematobium, S. intercalatum, S. bovis and Fasciola hepatica is the first step to maintain the life cycle of these parasites in laboratory in order to have biological material for the different studies, namely on the systematic biology and immunodiagnostic of schistosomosis and fasciolosis. According to the traditional method, the alga Oscillatoria formosa Bory (Cyanobacteria, principal food source for the snails, was cultivated in soil extract (Sampaio Xavier et al., 1968. However, it was sometimes very difficult to find the proper soil extract and the material was also contaminated by protozoa and fungi. In our work, using a new medium having as a base the Mineral Medium II (modified from Hughes et al., 1958 we found that O. formosa had a better growth response than in the soil extract medium. Snails fed on O. formosa reached three times the size of others at the same age, and they also reached sex maturity earlier, having more egg-masses per snail and, in addition, the rate of survival as well as the number of generations per year under laboratory conditions significantly increased. This culture was also easier to perform, and the axenic conditions easier to maintain.

  2. Growth and consumption of L-malic acid in wine-like medium by acclimated and non-acclimated cultures of Patagonian Oenococcus oeni strains.

    Science.gov (United States)

    Bravo-Ferrada, Bárbara Mercedes; Hollmann, Axel; Brizuela, Natalia; La Hens, Danay Valdés; Tymczyszyn, Elizabeth; Semorile, Liliana

    2016-09-01

    Five Oenococcus oeni strains, selected from spontaneous malolactic fermentation (MLF) of Patagonic Pinot noir wine, were assessed for their use as MLF starter cultures. After the individual evaluation of tolerance to some stress conditions, usually found in wine (pH, ethanol, SO2, and lysozyme), the behavior of the strains was analyzed in MLO broth with 14 % ethanol and pH 3.5 in order to test for the synergistic effect of high ethanol level and low pH and, finally, in a wine-like medium. Although the five strains were able to grow in MLO broth under low pH and/or high ethanol, they must be acclimated to grow in a wine-like medium. Additionally, glycosidase and tannase activities were evaluated, showing differences among the strains. The potential of the strains to ferment citrate was tested and two of the five strains showed the ability to metabolize this substrate. We did not detect the presence of genes encoding histidine, tyrosine descarboxylase, and putrescine carbamoyltransferase. All the strains tested exhibited good growth capacity and ability to consume L-malic acid in a wine-like medium after cell acclimation, and each of them showed a particular enzyme profile, which might confer different organoleptic properties to the wine.

  3. Surface characterization of insulin-coated Ti6Al4V medical implants conditioned in cell culture medium: An XPS study

    International Nuclear Information System (INIS)

    Shchukarev, Andrey; Malekzadeh, Behnosh Öhrnell; Ransjö, Maria; Tengvall, Pentti; Westerlund, Anna

    2017-01-01

    Highlights: • In the absence of FBS, chemically immobilized insulin layer remains intact; • The immobilized insulin expose hydrophobic domains outward the implant; • In the presence of FBS, a partial replacement of insulin occurs; • The immobilized insulin stabilizes the secondary structure of adsorbed proteins. - Abstract: Surface characterization of insulin-coated Ti6Al4V medical implants, after incubation in α-minimum essential medium (α-MEM), was done by X-ray photoelectron spectroscopy (XPS), in order to analyze the insulin behavior at the implant – α-MEM interface. In the absence of serum proteins in cell culture medium, the coated insulin layer remained intact, but experienced a time-dependent structural transformation exposing hydrophobic parts of the protein toward the solution. The presence of fetal bovine serum (FBS) in the medium resulted in partial substitution of insulin by serum proteins. In spite of some insulin release, the remaining coated layer demonstrated a direct surface effect by stabilizing the structure of protein competitors, and by supporting the accumulation of calcium and phosphate ions at the interface. A structurally stable protein layer with incorporated calcium and phosphate ions at the implant–tissue interface could be an important prerequisite for enhanced bone formation.

  4. Determination of the cause of the symptoms on yellow yam (Dioscorea cayenensis Lam.) leaf tissue and their eradication, enriching the culture medium and using techniques of meristem culture, thermo and chemotherapy on in vitro conditions

    International Nuclear Information System (INIS)

    Brenes Huertas, Mauricio

    2010-01-01

    Yams (Dioscorea spp) has been cultivated for exportation in Costa Rica, in North Huetar region. In vitro culture technique has been used for multiplying planting material for many advantages. However, cleaning of viruses that affect has been ineffective. Viruses such as: the potyvirus, potexvirus, cucumovirus . Methods like meristem culture, chemotherapy, thermotherapy and combinations of these have been used for the elimination of virus in plant species. The plants were evaluated in indexing assays, observing symptoms, serological methods and electron microscopy, among others. Other problems that have been affecting in vitro plant are deficient culture media in some nutrient. The presence of some abnormal characteristics in leaf tissue was determined whether have been caused by a virus or a nutritional deficiency in the culture medium. The presence of the virus has tried to find using ELISA and electron microscopy. Tests meristem culture, thermotherapy and chemotherapy have been made for the eradication of a possible virus; which have been assessed by observation of symptomatology and ELISA. The efficiency of the culture medium was evaluated to enrich it with nitrogen or excess iron. None of the suspected virus found in ELISA tests. Filaments are presumably viral particles were found through analysis of ultrastructure, as well as alterations in chloroplasts which indicated the presence of a pathogen or toxicity. Thermotherapy and chemotherapy with the concentration of 40 mg/L of ribavirin have been the most effective for the elimination of symptoms in virus eradication treatments. Assessments nutrient concentrations have shown that the differences between the various treatments used were undetectable. The symptoms presented were caused, according to the conclusions, by a virus which should preferably deal with thermotherapy. (author) [es

  5. Osteogenic medium is superior to growth factors in differentiation of human adipose stem cells towards bone-forming cells in 3D culture

    Directory of Open Access Journals (Sweden)

    L Tirkkonen

    2013-01-01

    Full Text Available Human adipose stem cells (hASCs have been recently used to treat bone defects in clinical practice. Yet there is a need for more optimal scaffolds and cost-effective approaches to induce osteogenic differentiation of hASCs. Therefore, we compared the efficiency of bone morphogenetic proteins (BMP-2 and BMP-7, vascular endothelial growth factor (VEGF, and osteogenic medium (OM for the osteo-induction of hASCs in 3D culture. In addition, growth factors were tested in combination with OM. Commercially available bioactive glass scaffolds (BioRestore and biphasic calcium phosphate granules (BoneCeramic were evaluated as prospective carriers for hASCs. Both biomaterials supported hASC-viability, but BioRestore resulted in higher cell number than BoneCeramic, whereas BoneCeramic supported more significant collagen production. The most efficient osteo-induction was achieved with plain OM, promoting higher alkaline phosphatase activity and collagen production than growth factors. In fact, treatment with BMP-2 or VEGF did not increase osteogenic differentiation or cell number significantly more than maintenance medium with either biomaterial. Moreover, BMP-7 treatment consistently inhibited proliferation and osteogenic differentiation of hASCs. Interestingly, there was no benefit from growth factors added to OM. This is the first study to demonstrate that OM enhances hASC-differentiation towards bone-forming cells significantly more than growth factors in 3D culture.

  6. Effect of culture medium on polymer production and temperature on recovery of polymer produced from newly identified Rhyzopus oryzae ST29

    Directory of Open Access Journals (Sweden)

    Tipparat Hongpattarakere

    2008-04-01

    Full Text Available Thermotolerant fungal isolate ST29 was identified by observing on cell morphology and molecular technique based on internal transcribed spacer (ITS gene to be Rhizopus oryzae. Among four culture media tested, the strain exhibited the highest growth in yeast malt extract (YM medium (4.87 g/l, followed by Sabouraud dextrose broth (SDB (4.25 g/l, potato dextrose broth (PDB (4.10 g/l and palm oil mill effluent (POME (3.29 g/l, respectively, after 4 days cultivation at 45oC. However, the strain was found to produce polymer only in POME medium at 45oC, but not in the three synthetic media tested. Effect of temperature on separation of the biopolymer produced by this fungal strain was studied by incubating the culture broth in water bath with temperatures in the range of room temperature to 70oC. The biopolymer was recovered by filtration, centrifugation, and precipitation by adding 4 volumes of 95% ethanol, then freeze-drying. These temperatures therefore had no influence on the biopolymer yields (5.58-5.78 g/l or on biomass yields (2.90-3.29 g/l.

  7. EFFECT OF CULTURE MEDIUM ON BACTERIOCIN PRODUCTION BY LACTOBACILLUS RHAMNOSUS HN001 AND LACTOBACILLUS REUTERI ATCC 53608

    Directory of Open Access Journals (Sweden)

    Aguilar-Uscanga B. R.

    2013-06-01

    Full Text Available The aim of this study was to evaluate the effect of media on bacteriocin production by Lactobacillus rhamnosus HN001 and Lactobacillus reuteri ATCC 53608 using three different media: YPM, YPF and MRS supplemented with glucose and K2HPO4. The optimum temperature was 37°C and initial pH 6.5. Bacteriocin-like substances produced by tested bacteria in MRS medium supplemented with glucose and K2HPO4 exhibited a broad antimicrobial spectrum determined by well diffusion assay against indicator bacteria Listeria monocytogenes, Lactobacillus sakei, Enterococcus faecium, Lactobacillus delbrueckii, Lactobacillus acidophilus, but no antimicrobial spectrum against E. coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, Bacillus cereus was detected. Bacteriocin was sensitive to protease IV, trypsin, pepsin and -amylases, but resistant to lipase. It was also resistant to detergents such as Tween 80, Triton-X and SDS. This bacteriocin was thermo-stable (resistant at 60°C, 90°C and 100°C for 30 min. Tested bacteria showed the best antimicrobial (bacteriocin-like activity after growth in MRS medium. Bacteriocin substances produced by tested bacteria showed promising thermo-stable technological properties.

  8. Salame elaborado com Lactobacillus plantarum fermentado em meio de cultura de plasma suíno Salami sausage prepared with Lactobacillus plantarum fermented in porcine plasma culture medium

    Directory of Open Access Journals (Sweden)

    Paulo Cezar Bastianello Campagnol

    2007-12-01

    Full Text Available Este trabalho teve por objetivo produzir uma cultura starter com uma cepa de Lactobacillus plantarum em um meio de cultura com plasma suíno e verificar a viabilidade de sua aplicação em salame. O meio de cultura foi preparado com plasma suíno e água destilada (1:1, pH 11,0. Após a esterilização, 300 mL foram adicionados de 400 mL de uma solução estéril de glicose e difosfato de potássio. A cepa de Lb. plantarum foi semeada no meio de cultura e submetida à fermentação em pH 7,0, durante 36 horas (100 rpm, 37 ± 0,1 °C. Ao alcançar a fase estacionária, a cultura foi centrifugada e ressuspendida em leite desnatado estéril, liofilizada e aplicada em salame. A influência do inóculo foi avaliada nas características microbiológicas, físico-químicas e sensoriais de salames. Os resultados encontrados foram comparados com tratamentos sem adição de cultura starter e com uma cultura comercial. O microrganismo Lb. plantarum teve um crescimento máximo de 9,82 Log UFC.mL-1, após 30 horas de fermentação. Os salames elaborados com a cultura starter produzida apresentaram uma queda de pH significativamente maior, e menor valor de atividade de água que os demais tratamentos. O microrganismo Lb. plantarum melhorou significativamente o sabor dos salames.The purpose of this work was to produce a starter culture with a strain of Lactobacillus plantarum in a porcine plasma culture medium and ascertain the viability of applying it in salami sausage. The culture medium was prepared with porcine plasma and distilled water (1:1, pH 11.0. After sterilization, 300 mL were added of 400 mL of a sterile solution of glucose and potassium diphosphate. The Lb. plantarum strain was inoculated into the culture medium and subjected to fermentation at pH 7.0 for 36 hours (100 rpm, 37 ± 0.1 °C. When the stationary phase was reached, the culture was centrifuged and resuspensed in sterile skimmed milk, lyophilized and applied to salami. An evaluation was

  9. Strain and culture medium optimization for production enhancement of prodiginines from marine-derived Streptomyces sp. GQQ-10

    Science.gov (United States)

    Li, Xueping; Zhang, Guojian; Zhu, Tianjiao; Li, Dehai; Gu, Qianqun

    2012-09-01

    A mutant (GQQ-M6) of a Sponge-Derived streptomyces sp. GQQ-10 obtained by UV-induced mutation was used for producing prodiginines (PGs). Single factor experiments and orthogonal array design (OAD) methods were employed for medium optimization. In the single factor method, the effects of soluble starch, glucose, soybean flour, yeast extract and sodium acetate on PGs production were investigated individually. In the subsequent OAD experiments, the concentrations of these 5 key nutritional components combined with salinity were further adjusted. The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain; OAD experiments offered a PGs yield of 61mg L-1, which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.

  10. Reusable Solid Rocket Motor - Accomplishment, Lessons, and a Culture of Success

    Science.gov (United States)

    Moore, D. R.; Phelps, W. J.

    2011-01-01

    The Reusable Solid Rocket Motor (RSRM) represents the largest solid rocket motor (SRM) ever flown and the only human-rated solid motor. High reliability of the RSRM has been the result of challenges addressed and lessons learned. Advancements have resulted by applying attention to process control, testing, and postflight through timely and thorough communication in dealing with all issues. A structured and disciplined approach was taken to identify and disposition all concerns. Careful consideration and application of alternate opinions was embraced. Focus was placed on process control, ground test programs, and postflight assessment. Process control is mandatory for an SRM, because an acceptance test of the delivered product is not feasible. The RSRM maintained both full-scale and subscale test articles, which enabled continuous improvement of design and evaluation of process control and material behavior. Additionally RSRM reliability was achieved through attention to detail in post flight assessment to observe any shift in performance. The postflight analysis and inspections provided invaluable reliability data as it enables observation of actual flight performance, most of which would not be available if the motors were not recovered. RSRM reusability offered unique opportunities to learn about the hardware. NASA is moving forward with the Space Launch System that incorporates propulsion systems that takes advantage of the heritage Shuttle and Ares solid motor programs. These unique challenges, features of the RSRM, materials and manufacturing issues, and design improvements will be discussed in the paper.

  11. COMPARATIVE EVALUTION OF CEPHALOSPORIN-C PRODUCTION IN SOLID STATE FERMENTATION AND SUBMERGED LIQUID CULTURE

    Directory of Open Access Journals (Sweden)

    Mahdi Rezazarandi

    2012-08-01

    Full Text Available The advantages of solid state fermentation (SSF utilization in producing enzymes & secondary metabolites have been shown, whereas, submerged liquid fermentation (SLF condition has the major usage in industrial production of antibiotics. As an antibiotic of B-lactams group, cephalosporin-C (CPC is indicated due to its wide effect and broad convention in treatment of infectious diseases. Regarding industrial production of CPC regularly done in SLF condition, we compared CPC production sum in SLF and SSF conditions. In this analysis, A. chrysogenum was employed, which was inoculated to SLF and SSF, while internal fermenter conditions were totally under control. After extraction of CPC, productions in two states of SLF and SSF were compared using the cylinder plate method. According to Antibiotic assay and production amount comparison, results expressed a ratio of development of production in SSF conditions to SLF conditions. Regarding previous studies on a solid state fermenter and its advantages, in this study, convenience of SSF conditions compared to SLF conditions was experimented. Also mentioning that maintaining the condition of solid state fermenter is more comfortable and practical than liquid state fermenter, using a solid based fermenter to produce antibiotics, especially CPC, can be appropriate. Considering appropriate control conditions of SSF to produce secondary metabolites, decrease in expenses, and increase of production, taking advantage of it in order to increase production parallel to modern methods, such as genetically manipulating CPC producing microorganisms are recommended to pharmacological industries. Also, to make this method applicable, further studies in industrial criterion seem necessary.

  12. Evaluation of the HB&L System for the Microbiological Screening of Storage Medium for Organ-Cultured Corneas.

    Science.gov (United States)

    Camposampiero, D; Grandesso, S; Zanetti, E; Mazzucato, S; Solinas, M; Parekh, M; Frigo, A C; Gion, M; Ponzin, D

    2013-01-01

    Aims. To compare HB&L and BACTEC systems for detecting the microorganisms contaminating the corneal storage liquid preserved at 31°C. Methods. Human donor corneas were stored at 4°C followed by preservation at 31°C. Samples of the storage medium were inoculated in BACTEC Peds Plus/F (aerobic microorganisms), BACTEC Plus Anaerobic/F (anaerobic microorganisms), and HB&L bottles. The tests were performed (a) after six days of storage, (b) end of storage, and (c) after 24 hours of preservation in deturgescent liquid sequentially. 10,655 storage and deturgescent media samples were subjected to microbiological control using BACTEC (6-day incubation) and HB&L (24-hour incubation) systems simultaneously. BACTEC positive/negative refers to both/either aerobic and anaerobic positives/negatives, whereas HB&L can only detect the aerobic microbes, and therefore the positives/negatives depend on the presence/absence of aerobic microorganisms. Results. 147 (1.38%) samples were identified positive with at least one of the two methods. 127 samples (134 identified microorganisms) were positive with both HB&L and BACTEC. 14 HB&L+/BACTEC- and 6 BACTEC+/HB&L- were identified. Sensitivity (95.5%), specificity (99.8%), and positive (90.1%) and negative predictive values (99.9%) were high with HB&L considering a 3.5% annual contamination rate. Conclusion. HB&L is a rapid system for detecting microorganisms in corneal storage medium in addition to the existing methods.

  13. Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

    Science.gov (United States)

    Lindborg, Beth A; Brekke, John H; Vegoe, Amanda L; Ulrich, Connor B; Haider, Kerri T; Subramaniam, Sandhya; Venhuizen, Scott L; Eide, Cindy R; Orchard, Paul J; Chen, Weili; Wang, Qi; Pelaez, Francisco; Scott, Carolyn M; Kokkoli, Efrosini; Keirstead, Susan A; Dutton, James R; Tolar, Jakub; O'Brien, Timothy D

    2016-07-01

    Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. ©AlphaMed Press.

  14. Clinical impact of culture-positive preservation fluid on solid organ transplantation: A systematic review and meta-analysis.

    Science.gov (United States)

    Oriol, Isabel; Sabé, N; Tebé, C; Veroux, M; Boin, I F S F; Carratalà, J

    2018-04-01

    Contamination of the preservation fluid (PF) used for donated organs is a potential source of post-transplant infection. However, the information on this issue is scarce. We therefore conducted a systematic review and meta-analysis to assess the incidence of culture-positive PF and its impact on solid organ transplant (SOT) recipients. Seventeen studies were identified and included. The overall incidence of culture-positive PF was 37% (95% CI: 27% to 49%), and the incidence of PF-related infections among SOT recipients with PF cultures that grew pathogenic microorganisms was 10% (95% CI: 7% to 15%). There were differences in the rates of infections due to pathogenic microorganisms between SOT recipients who received pre-emptive treatment and those who did not, but without statistical significance. The mortality rate among SOT recipients with PF-related infection was 35% (95% CI: 21% to 53%). In conclusion, although contamination of the PF of donated organs is frequent, the incidence of PF-related infection is relatively low. A closely clinical and microbiologic monitoring of the SOT recipient in case of culture-positive PF, regardless of the type of microorganism isolated might be do in order to establish a prompt diagnosis of PF-related infection. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Influence of serum extraction from the culture medium and of sublethal X-ray irradiation upon microvilli and invaginations of the membrane of Ehrlich ascites tumor cells in monolayer culture

    International Nuclear Information System (INIS)

    Laudenbach, G.; Pfab, R.; Hess, F.; Schachtschabel, D.O.

    1984-01-01

    In order to find out modifications of microvilli and invaginations, the cellular surfaces of Ehrlich ascites tumor cells in monolayer culture (basal medium of Eagle + 10% fetal calf serum) were investigated with the aid of electron-microscopic cross-sections. The tumor cells had been cultured without serum 24 hours prior to investigation or irradiated with 2 Gy. Morphometric evaluation after cell culture in a serum-free medium showed a reduced number of microvilli and a diminution of sections of microvilli. As already described before, a reduction of cell proliferation, of the microtubule-microfilament system, and of the endocytosis activity occurs under these serum-free conditions. The number of invaginations (related to a constant membrane part) was reduced by nearly 50% after serum extraction. Similarly to serum extraction, sublethal X-ray irradiation reduced the sections of microvilli, whereas the number of microvilli increased slightly. Contrary to the effect of serum extraction, the irradiated cells showed twice as many invaginations as the non-irradiated control cells. These differences in the surface structures are interpreted as a result of modified growth stimulations (+- serum) and radiogenic reparation processes. (orig.) [de

  16. Optimization of cultural conditions for the production of alpha amylase by aspergillus niger (btm-26) in solid state fermentation

    International Nuclear Information System (INIS)

    Abdullah, R.; Shaheen, N.; Iqtedar, M.; Naz, S.

    2014-01-01

    The present study deals with the isolation, screening and selection of native fungal strain for the alpha amylase production. Forty fungal strains were isolated from different soil samples. These strains were initially screened qualitatively on starch agar medium and quantitative screening was carried out in solid state fermentation. A strain of Aspergillus niger showing maximum production (432 +- 0.9 U/ml/min) of enzyme was selected and assigned the code BTM-26. The yield on various agricultural products, namely, coconut oil cake (COC), rice bran (RB), vegetable wastes or banana peel and wheat bran (WB) was compared. Wheat bran proved to be the best substrate for alpha amylase production. The effect of incubation temperature, initial pH, and inoculum size was investigated for the enzyme production. The maximum enzyme production was obtained at 30 degree C, pH 5, and inoculum size of 1 ml. The rate of fermentation was also studied and the highest yield of enzyme was obtained after 72 h of inoculation. Addition of 1.5% lactose as carbon source and 0.2% (NH/sub 4/)2SO/sub 4/ and 0.3% yeast extract as inorganic and organic nitrogen sources respectively gave enzyme production 990 +- 0.81 U/ml/min which reflects about 1.87 fold increase in alpha amylase production as compared to the medium containing wheat bran alone as substrate. (author)

  17. Comparison of BacT/Alert FAN and FAN Plus Bottles with Conventional Medium for Culturing Cerebrospinal Fluid.

    Science.gov (United States)

    Yoo, In Young; Chun, Sejong; Song, Dong Joon; Huh, Hee Jae; Lee, Nam Yong

    2016-11-01

    We compared the BacT/Alert system FAN and FAN Plus media to conventional media for culturing cerebrospinal fluid (CSF) with 2,545 samples. FAN/FAN Plus bottles showed better performance for isolating microorganisms in CSF than conventional media (positive rate, 7.2% [182/2,545] versus 3.1% [80/2,545]). The incremental recovery rate of Cryptococcus neoformans from FAN Plus bottles was higher than that from FAN bottles. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  18. Reusable Solid Rocket Motor - Accomplishments, Lessons, and a Culture of Success

    Science.gov (United States)

    Moore, Dennis R.; Phelps, Willie J.

    2011-01-01

    The Reusable Solid Rocket Motor represents the largest solid rocket motor ever flown and the only human rated solid motor. Each Reusable Solid Rocket Motor (RSRM) provides approximately 3-million lb of thrust to lift the integrated Space Shuttle vehicle from the launch pad. The motors burn out approximately 2 minutes later, separate from the vehicle and are recovered and refurbished. The size of the motor and the need for high reliability were challenges. Thrust shaping, via shaping of the propellant grain, was needed to limit structural loads during ascent. The motor design evolved through several block upgrades to increase performance and to increase safety and reliability. A major redesign occurred after STS-51L with the Redesigned Solid Rocket Motor. Significant improvements in the joint sealing systems were added. Design improvements continued throughout the Program via block changes with a number of innovations including development of low temperature o-ring materials and incorporation of a unique carbon fiber rope thermal barrier material. Recovery of the motors and post flight inspection improved understanding of hardware performance, and led to key design improvements. Because of the multidecade program duration material obsolescence was addressed, and requalification of materials and vendors was sometimes needed. Thermal protection systems and ablatives were used to protect the motor cases and nozzle structures. Significant understanding of design and manufacturing features of the ablatives was developed during the program resulting in optimization of design features and processing parameters. The project advanced technology in eliminating ozone-depleting materials in manufacturing processes and the development of an asbestos-free case insulation. Manufacturing processes for the large motor components were unique and safety in the manufacturing environment was a special concern. Transportation and handling approaches were also needed for the large

  19. Optimization of culture conditions for tannase production by Aspergillus sp. gm4 in solid state fermentation

    OpenAIRE

    Souza, Patrícia Nirlane da Costa; Universidade Federal de Lavras; Maia, Natália da Costa; Universidade Federal de Lavras; Guimarães, Luís Henrique Souza; Universidade de São Paulo; Resende, Mário Lúcio Vilela de; Universidade Federal de Lavras; Cardoso, Patrícia Gomes; Universidade Federal de Lavras

    2015-01-01

    The production of tannase by Aspergillus sp. GM4 under solid-state fermentation (SSF)  was investigated using different vegetables leaves such as mango, jamun, coffee and agricultural residues such as coffee husks, rice husks and wheat bran. Among substrates used jamun leaves yielded high tannase production. The Plackett-Burman design was conducted to evaluate the effects of 12 independent variables on the production of tannase under SSF using jamun leaves as substrate. Among these variables,...

  20. Size distribution of fullerenol nanoparticles in cell culture medium and their influence on antioxidative enzymes in Chinese hamster ovary cells

    Directory of Open Access Journals (Sweden)

    Srđenović Branislava U.

    2015-01-01

    Full Text Available Fullerenol (C60(OH24 nanoparticles (FNP have a significant role in biomedical research due to their numerous biological activities, some of which are cytoprotective and antioxidative properties. The aim of this study was to measure distribution of fullerenol nanoparticles and zeta potential in cell medium RPMI 1640 with 10% fetal bovine serum (FBS and to investigate the influence of FNP on Chinese hamster ovary cells (CHO-K1 survival, as well as to determine the activity of three antioxidative enzymes: superoxide-dismutase, glutathione-reductase and glutathione-S-transferase in mitomycin C-treated cell line. Our investigation implies that FNP, as a strong antioxidant, influence the cellular redox state and enzyme activities and thus may reduce cell proliferation, which confirms that FNP could be exploited for its use as a cytoprotective agent.[Projekat Ministarstva nauke Republike Srbije, br. III45005 i Pokrajinski Sekretarijat za nauku i tehnološki razvoj Vojvodine, grant number 114-451-2056/2011-01

  1. Stress- and sequence-dependent release into the culture medium of HIV-1 Nef produced in Saccharomyces cerevisiae.

    Science.gov (United States)

    Macreadie, I G; Castelli, L A; Lucantoni, A; Azad, A A

    1995-09-11

    We have produced human immunodeficiency virus type 1 (HIV-1) Nef (a myristylated 206-amino-acid protein) in Saccharomyces cerevisaie and shown that, while Nef is normally found as a predominantly intracellular protein, amounts up to 40 micrograms/ml of Nef are also released into the extracellular medium during stress. By electrophoretic (SDS-PAGE) analysis the extracellular Nef is indistinguishable from intracellular Nef. Conditions of stress that lead to the release of Nef include elevated levels of copper or magnesium ions or growth at elevated temperatures. This release appears to be dependent upon the N-terminal sequences of Nef, including the presence of a myristylation site. Our observations concerning Nef release in yeast suggest new ways in which the behaviour of Nef should be examined in order to gain further insights into the development of AIDS. If the release of Nef is important in the development of AIDS, our work reveals that Nef-associated symptoms may be reduced or delayed by reducing stresses, such as fevers.

  2. Study of the interaction of inorganic and organic compounds of cell culture medium with a Ti surface.

    Science.gov (United States)

    Burgos-Asperilla, L; García-Alonso, M C; Escudero, M L; Alonso, C

    2010-02-01

    The interaction between Ti and each component of Dulbecco's modified Eagle's medium was studied in depth using different techniques, such as the measurement of the corrosion potential, electrochemical impedance spectroscopy and polarization curves. The characterization of metal surfaces was carried out by scanning electron microscopy and X-ray photoelectron spectroscopy (XPS). The adsorption process of each component was studied using the quartz crystal balance (QCM). The QCM and XPS results reveal that the adsorption kinetics for phosphate and calcium ions is slow. However, the bovine serum albumin (BSA) totally covers the Ti surface rapidly. Because the passive film (titanium oxide) has acidic hydroxyl groups, the calcium ions would have a bridging effect on the electrostatic adsorption of phosphate ions as well as that of BSA. The polarization curves reveal that the adsorbed glucose permits the ionic diffusion of the oxygen to the electrode, while the BSA and fetal bovine serum (FBS) adsorbed after 7 days of immersion act as a diffusive barrier. The impedance measurement and data fitting to the electrical equivalent circuit model show that the resistance of the proteins/TiO(2) interface, for Ti immersed in FBS, is higher than those obtained for BSA, due to the proteins present in the solution as well as the fact that the adsorbed proteins on the surface are greater.

  3. Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test.

    Science.gov (United States)

    Riebeling, Christian; Schlechter, Katharina; Buesen, Roland; Spielmann, Horst; Luch, Andreas; Seiler, Andrea

    2011-06-01

    The embryonic stem cell test (EST) is a validated method to assess the developmental toxicity potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the mouse embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of bone morphogenetic protein-2, bone morphogenetic protein-4, activin A and ascorbic acid, embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Metabolism of sulfate-reducing bacteria and corrosion behavior of carbon steel in the continuous culturing medium; Renzoku baiyo baichichu ni okeru ryusan`en kangen no taisha to tansoko no fushoku kyodo

    Energy Technology Data Exchange (ETDEWEB)

    Baba, F.; Suzuki, T. [Ajinomoto Co. Inc., Tokyo (Japan); Seo, M. [Hokkaido University, Sapporo (Japan)

    1997-08-25

    Investigations were made on metabolism of sulfate-reducing bacteria and corrosion behavior of carbon steel in the continuous culturing medium. Sulfate-reducing bacteria were cultured for 50 days by supplying the culturing medium prepared to a prescribed chemical composition (containing Fe {sup 2+} at 0.01 mol/kg) at a rate of 10 cm {sup 3}/h, and drawing them out at the same rate. Test carbon steel pieces were immersed into this culturing medium. As a result, the following matters were clarified: the number of bacteria is maintained at more than 10 {sup 10}/cm{sup 3} after several days since inauguration of the immersion, with the bacteria stably producing H2S and FeS until the culturing is finished; comma-shaped bacteria which move actively and rod-shaped bacteria which do not move very actively exist in the culturing medium; a black film has been produced on surface of the test pieces throughout the culturing period, and satin-like corrosion was found underneath the surface; and weight increase of this film and weight decrease of the lower layer progress as the time lapses (the weight decrease of the lower layer has reached 40 mg/cm{sup 2} in 50 days). 28 refs., 8 figs., 1 tab.

  5. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

  6. Adding of ascorbic acid to the culture medium influences the antioxidant status and some biochemical parameters in the hen granulosa cells.

    Science.gov (United States)

    Capcarova, M; Kolesarova, A; Kalafova, A; Bulla, J; Sirotkin, A V

    2015-07-01

    The aim of the present study was to determine the activity of superoxide dismutase (SOD), total antioxidant status (TAS) of the hen granulosa cells, and selected biochemical parameters, including calcium, phosphorus, sodium, potassium, glucose, cholesterol, proteins, in the culture medium of granulosa cells after exposing them to ascorbic acid in vitro conditions. Ovarian granulosa cells of hens were incubated with various doses of ascorbic acid (E1 0.09 mg/ml, E2 0.13 mg/ml, E3 0.17 mg/ml, E4 0.33 mg/ml, E5 0.5 mg/ml). Ascorbic acid did not manifest antioxidant potential and higher doses of ascorbic acid (0.17; 0.33 and 0.5 mg/ml) decreased the activity of SOD in granulosa cells. Vitamin application resulted in a significantly (pascorbic acid might be involved in the regulation of selected biochemical and physiological processes in ovarian granulosa cells.

  7. In vitro culture of early secondary preantral follicles in hanging drop of ovarian cell-conditioned medium to obtain MII oocytes from outbred deer mice.

    Science.gov (United States)

    Choi, Jung Kyu; Agarwal, Pranay; He, Xiaoming

    2013-12-01

    The ovarian follicle (each contains a single oocyte) is the fundamental functional tissue unit of mammalian ovaries. In humans, it has been long held true that females are born with a maximum number of follicles (or oocytes) that are not only nonrenewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of ∼35. Therefore, it is of importance to isolate and bank ovarian follicles for in vitro culture to obtain fertilizable oocytes later, to preserve the fertility of professional women who may want to delay childbearing, young and unmarried women who may lose gonadal function because of exposure to environmental/occupational hazards or aggressive medical treatments, such as radiation and chemotherapy, and even endangered species and breeds. Although they contributed significantly to the understanding of follicle science and biology, most studies reported to date on this topic were done using the man-made, unnatural inbred animal species. It was found in this study that the conventional two-dimensional microliter drop and three-dimensional hanging drop (HD) methods, reported to be effective for in vitro culture of preantral follicles from inbred mice, are not directly transferrable to outbred deer mice. Therefore, a modified HD method was developed in this study to achieve a much higher (>5 times compared to the best conventional methods) percentage of developing early secondary preantral follicles from the outbred mice to the antral stage, for which, the use of an ovarian cell-conditioned medium and multiple follicles per HD were identified to be crucial. It was further found that the method for in vitro maturation of oocytes in antral follicles obtained by in vitro culture of preantral follicles could be very different from that for oocytes in antral follicles obtained by hormone stimulation in vivo. Therefore, this study should provide important guidance for establishing effective protocols of in vitro follicle

  8. Interaction with culture medium components, cellular uptake and intracellular distribution of cobalt nanoparticles, microparticles and ions in Balb/3T3 mouse fibroblasts.

    Science.gov (United States)

    Sabbioni, Enrico; Fortaner, Salvador; Farina, Massimo; Del Torchio, Riccardo; Petrarca, Claudia; Bernardini, Giovanni; Mariani-Costantini, Renato; Perconti, Silvia; Di Giampaolo, Luca; Gornati, Rosalba; Di Gioacchino, Mario

    2014-02-01

    The mechanistic understanding of nanotoxicity requires the physico-chemical characterisation of nanoparticles (NP), and their comparative investigation relative to the corresponding ions and microparticles (MP). Following this approach, the authors studied the dissolution, interaction with medium components, bioavailability in culture medium, uptake and intracellular distribution of radiolabelled Co forms (CoNP, CoMP and Co(2+)) in Balb/3T3 mouse fibroblasts. Co(2+) first saturates the binding sites of molecules in the extracellular milieu (e.g., albumin and histidine) and on the cell surface. Only after saturation, Co(2+) is actively uptaken. CoNP, instead, are predicted to be internalised by endocytosis. Dissolution of Co particles allows the formation of Co compounds (CoNP-rel), whose mechanism of cellular internalisation is unknown. Co uptake (ranking CoMP > CoNP > Co(2+)) reached maximum at 4 h. Once inside the cell, CoNP spread into the cytosol and organelles. Consequently, massive amounts of Co ions and CoNP-rel can reach subcellular compartments normally unexposed to Co(2+). This could explain the fact that the nuclear and mitochondrial Co concentrations resulted significantly higher than those obtained with Co(2+).

  9. Cytotoxicity of cancer HeLa cells sensitivity to normal MCF10A cells in cultivations with cell culture medium treated by microwave-excited atmospheric pressure plasmas

    Science.gov (United States)

    Takahashi, Yohei; Taki, Yusuke; Takeda, Keigo; Hashizume, Hiroshi; Tanaka, Hiromasa; Ishikawa, Kenji; Hori, Masaru

    2018-03-01

    Cytotoxic effects of human epithelial carcinoma HeLa cells sensitivity to human mammary epithelial MCF10A cells appeared in incubation with the plasma-activated medium (PAM), where the cell culture media were irradiated with the hollow-shaped contact of a continuously discharged plasma that was sustained by application of a microwave power under Ar gas flow at atmospheric pressure. The discharged plasma had an electron density of 7  ×  1014 cm-3. As the nozzle exit to the plasma source was a distance of 5 mm to the medium, concentrations of 180 µM for H2O2 and 77 µM for NO2- were generated in the PAM for 30 s irradiation, resulting in the control of irradiation periods for aqueous H2O2 with a generation rate of 6.0 µM s-1, and nitrite ion (NO2- ) with a rate of 2.2 µM s-1. Effective concentrations of H2O2 and NO2- for the antitumor effects were revealed in the microwave-excited PAM, with consideration of the complicated reactions at the plasma-liquid interfaces.

  10. Optimisation of critical medium components and culture conditions for enhanced biomass and lipid production in the oleaginous diatom Navicula phyllepta: a statistical approach.

    Science.gov (United States)

    Sabu, Sanyo; Singh, Isaac Sarojini Bright; Joseph, Valsamma

    2017-12-01

    Diatoms hold great promise as potential sources of biofuel production. In the present study, the biomass and lipid production in the marine diatom Navicula phyllepta, isolated from Cochin estuary, India and identified as a potential biodiesel feedstock, were optimized using Plackett-Burman (PB) statistical experimental design followed by central composite design (CCD) and response surface methodology (RSM). The growth analyses of the isolate in different nitrogen sources, salinities and five different enriched sea water media showed the best growth in the cheapest medium with minimum components using urea as nitrogen source at salinity between 25 and 40 g kg -1 . Plackett-Burman experimental analyses for screening urea, sodium metasilicate, sodium dihydrogen phosphate, ferric chloride, salinity, temperature, pH and agitation influencing lipid and biomass production showed that silicate and temperature had a positive coefficient on biomass production, and temperature had a significant positive coefficient, while urea and phosphate showed a negative coefficient on lipid content. A 2 4 factorial central composite design (FCCD) was used to optimize the concentration of the factors selected. The optimized media resulted in 1.62-fold increase (64%) in biomass (1.2 ± 0.08 g L -1 ) and 1.2-fold increase (22%) in estimated total lipid production (0.11 ± 0.003 g L -1 ) compared to original media within 12 days of culturing. A significantly higher biomass and lipid production in the optimized medium demands further development of a two-stage strategy of biomass production followed by induction of high lipid production under nutrient limitation or varying culture conditions for large-scale production of biodiesel from the marine diatom.

  11. The effect of inactivated Lactobacillus LB fermented culture medium on symptom severity: observational investigation in 297 patients with diarrhea-predominant irritable bowel syndrome.

    Science.gov (United States)

    Tarrerias, A L; Costil, V; Vicari, F; Létard, J C; Adenis-Lamarre, P; Aisène, A; Batistelli, D; Bonnaud, G; Carpentier, S; Dalbiès, P; Ecuer, S; Etienne, J; Fantoli, M; Grunberg, B; Lannoy, P; Lapuelle, J; Margulies, A; Neumeier, M; Rouillon, J M; Schmets, L; Pingannaud, M P; Coulom, P; Kholer, F; Canard, J M

    2011-01-01

    Little is known about the intensity of symptoms of diarrhea-predominant IBS (IBS-D) or the consequences of the disease on patients' health-related quality of life (HRQOL). This observational investigation assessed the symptoms (abdominal pain, bloating, number of stools per day, and stool consistency), impact on HRQOL, and consequence on anal continence in 297 patients with IBS-D before and after 1 month of probiotic treatment with Lacteol (inactivated Lactobacillus LB plus fermented culture medium). Functional assessment using a standardized visual analogue scale in order to quantify abdominal pain, bloating, and quality of life before and after 1 month of treatment with 2 capsules/day of Lacteol. The number of symptomatic days per week, number of stools, consistency of stools, secondary fecal incontinence rate, and potential trigger effect of food were quantified. A χ2 test was used to compare qualitative data and the variance of quantitative criteria was analyzed. The pain score decreased from 4.46±0.15 on a scale of 0-10 before treatment to 2.8±0.14 after treatment (pspices, and 4% to sugar. This observational investigation shed new light on patients with IBS-D, the HRQOL of which is altered by a fecal incontinence rate twice as high as that of the general population. Correlation with diet is confirmed by 1 out of 2 patients reporting poor tolerance of fiber and dairy products. Nutritional management should thus be part of these patients' treatment. Inactivated Lactobacillus LB plus fermented culture medium is a probiotic drug that has been used by physicians for a long time to treat patients with diarrhea. Strongly concentrated, it has no side effects and seems to help these patients. Due to a strong placebo effect in patients with this pathology, however, a controlled study is necessary to confirm this result. Copyright © 2011 S. Karger AG, Basel.

  12. A Clostridium Group IV Species Dominates and Suppresses a Mixed Culture Fermentation by Tolerance to Medium Chain Fatty Acids Products

    Science.gov (United States)

    Andersen, Stephen J.; De Groof, Vicky; Khor, Way Cern; Roume, Hugo; Props, Ruben; Coma, Marta; Rabaey, Korneel

    2017-01-01

    A microbial community is engaged in a complex economy of cooperation and competition for carbon and energy. In engineered systems such as anaerobic digestion and fermentation, these relationships are exploited for conversion of a broad range of substrates into products, such as biogas, ethanol, and carboxylic acids. Medium chain fatty acids (MCFAs), for example, hexanoic acid, are valuable, energy dense microbial fermentation products, however, MCFA tend to exhibit microbial toxicity to a broad range of microorganisms at low concentrations. Here, we operated continuous mixed population MCFA fermentations on biorefinery thin stillage to investigate the community response associated with the production and toxicity of MCFA. In this study, an uncultured species from the Clostridium group IV (related to Clostridium sp. BS-1) became enriched in two independent reactors that produced hexanoic acid (up to 8.1 g L−1), octanoic acid (up to 3.2 g L−1), and trace concentrations of decanoic acid. Decanoic acid is reported here for the first time as a possible product of a Clostridium group IV species. Other significant species in the community, Lactobacillus spp. and Acetobacterium sp., generate intermediates in MCFA production, and their collapse in relative abundance resulted in an overall production decrease. A strong correlation was present between the community composition and both the hexanoic acid concentration (p = 0.026) and total volatile fatty acid concentration (p = 0.003). MCFA suppressed species related to Clostridium sp. CPB-6 and Lactobacillus spp. to a greater extent than others. The proportion of the species related to Clostridium sp. BS-1 over Clostridium sp. CPB-6 had a strong correlation with the concentration of octanoic acid (p = 0.003). The dominance of this species and the increase in MCFA resulted in an overall toxic effect on the mixed community, most significantly on the Lactobacillus spp., which resulted in a decrease in total

  13. Structural investigation and response surface optimisation for improvement of kefiran production yield from a low-cost culture medium.

    Science.gov (United States)

    Ghasemlou, Mehran; Khodaiyan, Faramarz; Jahanbin, Kambiz; Gharibzahedi, Seyed Mohammad Taghi; Taheri, Salman

    2012-07-15

    Kefiran, a water-soluble heteropolysaccharide with molecular weight of 1.35×10(6)Da and a specific optical rotation of +64° (c 1.0, H2O), was isolated from kefir grains grown in cheese whey and further purified through DEAE-Sepharose XK26. Response surface methodology was employed to optimise the culture conditions for kefiran production from kefir grains to be lactose concentration 67 g/l, yeast extract 13g/l, pH 5.7 and temperature 24°C. Intrinsic viscosity was 5.84 dl/g using the Huggins extrapolation and 5.53 dl/g using the Kramer extrapolation. Monosaccharide analysis revealed that kefiran is composed of glucose (Glc) and galactose (Gal) in a relative molar ratio of 1.0:1.1. Its structural features were elucidated by a combination of FT-IR, methylation and GC-MS analysis, periodate oxidation-Smith degradation, partial acid hydrolysis and NMR spectroscopy ((1)H, (13)C and HMBC). The data obtained indicated that kefiran possessed a backbone of (1→6)-linked Glc, (1→3)-linked Gal, (1→4)-linked Gal, (1→4)-linked Glc and (1→2,6)-linked Gal, with a branch attached to O-2 of Gal residues and terminated with Glc residues. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Mouse preimplantation embryo responses to culture medium osmolarity include increased expression of CCM2 and p38 MAPK activation

    Directory of Open Access Journals (Sweden)

    Watson Andrew J

    2007-01-01

    Full Text Available Abstract Background Mechanisms that confer an ability to respond positively to environmental osmolarity are fundamental to ensuring embryo survival during the preimplantation period. Activation of p38 mitogen-activated protein kinase (MAPK occurs following exposure to hyperosmotic treatment. Recently, a novel scaffolding protein called Osmosensing Scaffold for MEKK3 (OSM was linked to p38 MAPK activation in response to sorbitol-induced hypertonicity. The human ortholog of OSM is cerebral cavernous malformation 2 (CCM2. The present study was conducted to investigate whether CCM2 is expressed during mouse preimplantation development and to determine whether this scaffolding protein is associated with p38 MAPK activation following exposure of preimplantation embryos to hyperosmotic environments. Results Our results indicate that Ccm2 along with upstream p38 MAPK pathway constituents (Map3k3, Map2k3, Map2k6, and Map2k4 are expressed throughout mouse preimplantation development. CCM2, MAP3K3 and the phosphorylated forms of MAP2K3/MAP2K6 and MAP2K4 were also detected throughout preimplantation development. Embryo culture in hyperosmotic media increased p38 MAPK activity in conjunction with elevated CCM2 levels. Conclusion These results define the expression of upstream activators of p38 MAPK during preimplantation development and indicate that embryo responses to hyperosmotic environments include elevation of CCM2 and activation of p38 MAPK.

  15. Production of Gymnemic Acid Depends on Medium, Explants, PGRs, Color Lights, Temperature, Photoperiod, and Sucrose Sources in Batch Culture of Gymnema sylvestre

    Directory of Open Access Journals (Sweden)

    A. Bakrudeen Ali Ahmed

    2012-01-01

    Full Text Available Gymnema sylvestre (R.Br. is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA. The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L and KN (0.5 mg/L. Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w. Maximum GA production (58.28 mg/g d.w was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures of G. sylvestre. The factors optimized for in vitro production of GA during the present study can successfully be employed for their large-scale production in bioreactors.

  16. Liquid culture enhances diagnosis of patients with milder forms of non-tuberculous mycobacterial lung disease.

    Science.gov (United States)

    Lee, H; Han, J-H; Park, H Y; Jeon, K; Huh, H J; Ki, C-S; Lee, N Y; Koh, W-J

    2017-03-01

    To evaluate the proportion and clinical characteristics of patients with non-tuberculous mycobacteria (NTM) lung disease diagnosed based on positive culture results in liquid medium only. We reviewed the medical records of 978 patients diagnosed with NTM lung disease. All clinical samples were cultured in both solid and liquid media. Of the 978 patients, 111 (11.3%) were culture-positive in liquid medium only (liquid culture group), and 867 (88.7%) (solid culture group) on solid medium, regardless of the culture results in liquid medium. At the time of diagnosis, the liquid culture group was less likely than the solid culture group to have haemoptysis (11.7% vs. 20.0%, P = 0.04), positive sputum smear for acid-fast bacilli (14.4% vs. 50.2%, P disease (3.6% vs. 14.6%, P = 0.001). During the median follow-up period of 28.9 months (interquartile range 19.1-41.6), the proportion of patients requiring antibiotic treatment was lower in the liquid culture group than in the solid culture group (44.1% vs. 61.6%, P culture is helpful in the diagnosis of patients with less severe forms of NTM lung disease.

  17. Optimization of fermentation parameters to study the behavior of selected lactic cultures on soy solid state fermentation.

    Science.gov (United States)

    Rodríguez de Olmos, A; Bru, E; Garro, M S

    2015-03-02

    The use of solid fermentation substrate (SSF) has been appreciated by the demand for natural and healthy products. Lactic acid bacteria and bifidobacteria play a leading role in the production of novel functional foods and their behavior is practically unknown in these systems. Soy is an excellent substrate for the production of functional foods for their low cost and nutritional value. The aim of this work was to optimize different parameters involved in solid state fermentation (SSF) using selected lactic cultures to improve soybean substrate as a possible strategy for the elaboration of new soy food with enhanced functional and nutritional properties. Soy flour and selected lactic cultures were used under different conditions to optimize the soy SSF. The measured responses were bacterial growth, free amino acids and β-glucosidase activity, which were analyzed by applying response surface methodology. Based on the proposed statistical model, different fermentation conditions were raised by varying the moisture content (50-80%) of the soy substrate and temperature of incubation (31-43°C). The effect of inoculum amount was also investigated. These studies demonstrated the ability of selected strains (Lactobacillus paracasei subsp. paracasei and Bifidobacterium longum) to grow with strain-dependent behavior on the SSF system. β-Glucosidase activity was evident in both strains and L. paracasei subsp. paracasei was able to increase the free amino acids at the end of fermentation under assayed conditions. The used statistical model has allowed the optimization of fermentation parameters on soy SSF by selected lactic strains. Besides, the possibility to work with lower initial bacterial amounts to obtain results with significant technological impact was demonstrated. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William's E Medium followed by Hepatocyte Differentiation Inducer Treatment.

    Directory of Open Access Journals (Sweden)

    Minoru Tomizawa

    Full Text Available Hepatocyte differentiation inducer (HDI lacks both glucose and arginine, but is supplemented with galactose and ornithine, and is added together with other reagents such as apoptosis inhibitor and oncostatin M. Although human induced pluripotent stem (iPS cells initiate hepatocyte differentiation, most die within 7 days. In this study, we investigated both HDI and conventional media for their potential to improve cell survival.201B7 iPS cells were cultured in conventional media. This consisted of three cycles of 5-day culture in William's E (WE medium, followed by a 2-day culture in HDI.Expression levels of α-feto protein (AFP were higher in cells cultured in WE and in Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DF12. 201B7 cells expressed the highest AFP and albumin (ALB when cultured in HDI for 2 days following 7-day culture in WE. After three cycles of 5-day culture in WE followed by 2 days in HDI, 201B7 cells expressed AFP and ALB 54 ± 2.3 (average ± standard deviation and 73 ± 15.1 times higher, respectively, than those cultured in ReproFF (feeder-free condition.201B7 cells survived culture in WE for 7 days followed HDI for 2 days. After three cycles of culture under these conditions, hepatocyte differentiation was enhanced, as evidenced by increased AFP and ALB expression.

  19. Comparison between point-of-care dermatophyte test medium and mycology laboratory culture for diagnosis of dermatophytosis in dogs and cats.

    Science.gov (United States)

    Kaufmann, Ronnie; Blum, Shlomo E; Elad, Daniel; Zur, Gila

    2016-08-01

    Point-of-care Dermatophyte Test Medium (PoC-DTM) is a diagnostic procedure to rule in/rule out dermatophytosis in veterinary clinics. To evaluate the performance of PoC-DTM in the clinic compared to DTM plate culture in a mycology laboratory and to compare results obtained by general practitioners and referral clinicians. Hair samples were collected from 47 cats and 54 dogs with suspected dermatophytosis and from nine healthy controls (seven cats and two dogs). This was a multicentre blinded study. In one group (65 suspected cases, 9 healthy controls), PoC-DTM results were evaluated by clinicians in a referral clinic (SP group) who examined the colony morphology macroscopically and microscopically. In the other group (36 suspected cases) PoC-DTM results were evaluated by clinicians from general practice for colour change only, with no macroscopic or microscopic examination (GP group). All hair samples were also cultured on DTM plates in a mycology laboratory. Laboratory culture was considered the gold standard for comparison. Agreements between tests were 97% (two false positive; κ = 0.839) and 80.6% (five false positives and two false negatives; κ = 0.466) in the SP and GP groups, respectively. This difference between groups was significant (P = 0.024). When applying macroscopic and microscopic evaluation of the colony, PoC-DTM is accurate for diagnosing dermatophytes with only a 3% chance of error. However, when macroscopic and microscopic examination is not included there is significant (19.4%) chance for an incorrect diagnosis. © 2016 ESVD and ACVD.

  20. Addition of granulosa cell mass to the culture medium of oocytes derived from early antral follicles increases oocyte growth, ATP content, and acetylation of H4K12.

    Science.gov (United States)

    Sugiyama, Miyako; Sumiya, Mei; Shirasuna, Koumei; Kuwayama, Takehito; Iwata, Hisataka

    2016-12-01

    The main aim of the present study was to examine the hypothesis that an increase in the number of granulosa cells surrounding developing bovine oocytes results in both high ATP levels and an increase in the acetylation level of H4K12 in oocytes grown in vitro. Oocyte-granulosa cell complexes (OGCs) were collected from early antral follicles (EAFs, 0.4-0.7 mm in diameter), and individually cultured on 96-well plates with or without additional granulosa cell mass that had been prepared from other OGCs. After 16 days of culture, we examined: (i) the rate of antrum formation of the OGCs; (ii) the diameter, maturation, and fertilization rate of the oocytes; and (iii) the ATP content and acetylation level of H4K12 in the oocytes grown in vitro. Granulosa cell mass added to the culture medium contributed to the development of OGCs with a higher rate of antrum formation and oocyte growth. Furthermore, the addition of granulosa cells increased the ATP content and acetylation level of H4K12 in oocytes grown in vitro compared with those developed without addition of granulosa cells. In addition, there was a positive correlation between the ATP content in oocytes grown in vitro and the number of granulosa cells in the corresponding OGCs. The results suggest that granulosa cells play a role not only in the development of OGCs and the growth of oocytes, but also in the determination of ATP content and the acetylation of H4K12 in the oocytes developed in vitro.

  1. Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

    Directory of Open Access Journals (Sweden)

    Takefuji Haruka

    2016-01-01

    Full Text Available Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-substrate, 95 h, and when this ratio was 200:1, glucoamylase showed maximum activity (180 U/g-substrate, 95 h.

  2. Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

    OpenAIRE

    Takefuji Haruka; Ninomiya Junko; Morita Hiroshi

    2016-01-01

    Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-subs...

  3. Anti-HCV effect of Lentinula edodes mycelia solid culture extracts and low-molecular-weight lignin.

    Science.gov (United States)

    Matsuhisa, Koji; Yamane, Seiji; Okamoto, Toru; Watari, Akihiro; Kondoh, Masuo; Matsuura, Yoshiharu; Yagi, Kiyohito

    2015-06-19

    Lentinula edodes mycelia solid culture extract (MSCE) contains several bioactive molecules, including some polyphenolic compounds, which exert immunomodulatory, antitumor, and hepatoprotective effects. In this study, we examined the anti-hepatitis C virus (HCV) activity of MSCE and low-molecular-weight lignin (LM-lignin), which is the active component responsible for the hepatoprotective effect of MSCE. Both MSCE and LM-lignin inhibited the entry of two HCV pseudovirus (HCVpv) types into Huh7.5.1 cells. LM-lignin inhibited HCVpv entry at a lower concentration than MSCE and inhibited the entry of HCV particles in cell culture (HCVcc). MSCE also inhibited HCV subgenome replication. LM-lignin had no effect on HCV replication, suggesting that MSCE contains additional active substances. We demonstrate here for the first time the anti-HCV effects of plant-derived LM-lignin and MSCE. The hepatoprotective effect of LM-lignin suggests that lignin derivatives, which can be produced in abundance from existing plant resources, may be effective in the treatment of HCV-related diseases. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Safety objectives and basic design for surface centres for long-term storage of solid radioactive waste with short or medium half-life and low or medium specific activity

    International Nuclear Information System (INIS)

    1984-06-01

    RFS or Regles Fondamentales de Surete (Basic Safety Rules) applicable to certain types of nuclear facilities lay down requirements with which compliance, for the type of facilities and within the scope of application covered by the RFS, is considered to be equivalent to compliance with technical French regulatory practice. The object of the RFS is to take advantage of standardization in the field of safety, while allowing for technical progress in that field. They are designed to enable the operating utility and contractors to know the rules pertaining to various subjects which are considered to be acceptable by the Service Central de Surete des Installations Nucleaires, or the SCSIN (Central Department for the Safety of Nuclear Facilities). These RFS should make safety analysis easier and lead to better understanding between experts and individuals concerned with the problems of nuclear safety. The SCSIN reserves the right to modify, when considered necessary, any RFS and specify, if need be, the terms under which a modification is deemed retroactive. The role of this RFS is to define the safety objectives and the basic design philosophy for surface centres for long-term storage of packages of radioactive waste with short or medium half-life and with low or medium specific activity

  5. Estimation of track registration efficiency in solution medium and study of gamma irradiation effects on the bulk-etch rate and the activation energy for bulk etching of CR-39 (DOP) Solid State Nuclear Track Detector

    International Nuclear Information System (INIS)

    Kalsi, P.C.

    2010-01-01

    The fission track registration efficiency of diethylene glycol bis allyl carbonate (dioctyl phthalate doped) (CR-39 (DOP)) solid state nuclear track detector (SSNTD) in solution medium (K wet ) has been experimentally determined and is found to be (9.7 ± 0.5).10 -4 cm. This is in good agreement with the values of other SSNTDs. The gamma irradiation effects in the dose range of 50.0-220.0 kGy on the bulk etch rate, V b and the activation energy for bulk etching, E of this solid state nuclear track detector (SSNTD) have also been studied. It is observed that the bulk etch rates increase and the activation energies for bulk etching decrease with the increase in gamma dose. These results have been explained on the basis of scission of the detector due to gamma irradiation

  6. Effect of culture levels, ultrafiltered retentate addition, total solid levels and heat treatments on quality improvement of buffalo milk plain set yoghurt.

    Science.gov (United States)

    Yadav, Vijesh; Gupta, Vijay Kumar; Meena, Ganga Sahay

    2018-05-01

    Studied the effect of culture (2, 2.5 and 3%), ultrafiltered (UF) retentate addition (0, 11, 18%), total milk solids (13, 13.50, 14%) and heat treatments (80 and 85 °C/30 min) on the change in pH and titratable acidity (TA), sensory scores and rheological parameters of yoghurt. With 3% culture levels, the required TA (0.90% LA) was achieved in minimum 6 h incubation. With an increase in UF retentate addition, there was observed a highly significant decrease in overall acceptability, body and texture and colour and appearance scores, but there was highly significant increase in rheological parameters of yoghurt samples. Yoghurt made from even 13.75% total solids containing nil UF retentate was observed to be sufficiently firm by the sensory panel. Most of the sensory attributes of yoghurt made with 13.50% total solids were significantly better than yoghurt prepared with either 13 or 14% total solids. Standardised milk heated to 85 °C/30 min resulted in significantly better overall acceptability in yoghurt. Overall acceptability of optimised yoghurt was significantly better than a branded market sample. UF retentate addition adversely affected yoghurt quality, whereas optimization of culture levels, totals milk solids and others process parameters noticeably improved the quality of plain set yoghurt with a shelf life of 15 days at 4 °C.

  7. Immunohistochemical expression of stem cell, endothelial cell, and chemosensitivity markers in primary glioma spheroids cultured in serum-containing and serum-free medium

    DEFF Research Database (Denmark)

    Christensen, Karina; Aaberg-Jessen, Charlotte; Andersen, Claus

    2010-01-01

    To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas.......To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas....

  8. Characterization of the Embryogenic Tissue of the Norway Spruce Including a Transition Layer between the Tissue and the Culture Medium by Magnetic Resonance Imaging

    Directory of Open Access Journals (Sweden)

    Kořínek R.

    2017-02-01

    Full Text Available The paper describes the visualization of the cells (ESEs and mucilage (ECMSN in an embryogenic tissue via magnetic resonance imaging (MRI relaxometry measurement combined with the subsequent multi-parametric segmentation. The computed relaxometry maps T1 and T2 show a thin layer (transition layer between the culture medium and the embryogenic tissue. The ESEs, mucilage, and transition layer differ in their relaxation times T1 and T2; thus, these times can be used to characterize the individual parts within the embryogenic tissue. The observed mean values of the relaxation times T1 and T2 of the ESEs, mucilage, and transition layer are as follows: 1469 ± 324 and 53 ± 10 ms, 1784 ± 124 and 74 ± 8 ms, 929 ± 164 and 32 ± 4.7 ms, respectively. The multi-parametric segmentation exploiting the T1 and T2 relaxation times as a classifier shows the distribution of the ESEs and mucilage within the embryogenic tissue. The discussed T1 and T2 indicators can be utilized to characterize both the growth-related changes in an embryogenic tissue and the effect of biotic/abiotic stresses, thus potentially becoming a distinctive indicator of the state of any examined embryogenic tissue.

  9. Energy-based culture medium design for biomanufacturing optimization: A case study in monoclonal antibody production by GS-NS0 cells.

    Science.gov (United States)

    Quiroga-Campano, Ana L; Panoskaltsis, Nicki; Mantalaris, Athanasios

    2018-03-02

    Demand for high-value biologics, a rapidly growing pipeline, and pressure from competition, time-to-market and regulators, necessitate novel biomanufacturing approaches, including Quality by Design (QbD) principles and Process Analytical Technologies (PAT), to facilitate accelerated, efficient and effective process development platforms that ensure consistent product quality and reduced lot-to-lot variability. Herein, QbD and PAT principles were incorporated within an innovative in vitro-in silico integrated framework for upstream process development (UPD). The central component of the UPD framework is a mathematical model that predicts dynamic nutrient uptake and average intracellular ATP content, based on biochemical reaction networks, to quantify and characterize energy metabolism and its adaptive response, metabolic shifts, to maintain ATP homeostasis. The accuracy and flexibility of the model depends on critical cell type/product/clone-specific parameters, which are experimentally estimated. The integrated in vitro-in silico platform and the model's predictive capacity reduced burden, time and expense of experimentation resulting in optimal medium design compared to commercially available culture media (80% amino acid reduction) and a fed-batch feeding strategy that increased productivity by 129%. The framework represents a flexible and efficient tool that transforms, improves and accelerates conventional process development in biomanufacturing with wide applications, including stem cell-based therapies. Copyright © 2018. Published by Elsevier Inc.

  10. Recovery of mango plants with antrachnose resistance following mutation induction and selection in vitro with the culture filtrate of Colletotrichum gloesporoides Penz

    Energy Technology Data Exchange (ETDEWEB)

    Litz, R. E. [Tropical Research and Education Center, University of Florida, Homestead, FL (United States)

    2009-05-15

    Embryogenic mango cultures of three cultivars on semi solid medium were irradiated at 100 Gy: monoembryonic ‘Tommy Atkins’ and ‘Keitt’ and polyembryonic ‘Hindi be Sennara’. Two weeks after irradiation, cultures were inoculated into liquid maintenance medium containing 10% (v/v) culture filtrate of Colletotrichum gloeosporioides Penz. Following two weeks of exposure to culture filtrate, the embryogenic cultures were sub-cultured onto semi solid maintenance medium. Living pro-embryonic masses were manually separated from necrotic tissue four weeks later and were transferred onto semi solid maintenance medium. Somatic embryos were recovered and their shoots have been rescued by ex vitro grafting. Field plantings were established in early 2005; however, the results are inconclusive at the time of writing. (author)

  11. Regeneration from irradiated avocado (Persea americana Mill.) embryogenic cultures

    Energy Technology Data Exchange (ETDEWEB)

    Witjaksono, Y.; Nugraheni, U. K.; Hoesen, D. H. [Plant Cell and Tissue Culture Laboratory, Bogor Botanic Garden, Bogor (Indonesia); Litz, R. E. [Tropical Research and Education Center, University of Florida, Homestead, FL (United States)

    2009-05-15

    Somatic embryogenesis was induced from zygotic embryos excised from immature avocado fruit from selected genotypes grown in the highlands of Cisarua, West Bogor, Indonesia. The proembryonic masses developed first on semi-solid medium and were then transferred to liquid cultures for proliferation. The embryogenic masses were then irradiated at 9, 18 and 35 Gy using a {sup 60}Co irradiation source. 3 sub-cultures in liquid medium ensured adequate proliferation prior to transfer to fresh development medium. After 1-3 months, somatic embryos with more than 0.5 cm in diameter were transferred to a germination medium, while the smaller somatic embryos (<0.5 cm in diameter) were sub-cultured one more time for additional growth. After 1-2 months on germination medium, plantlets were transferred individually to new medium. (author)

  12. Culture.

    Science.gov (United States)

    Smith, Timothy B; Rodríguez, Melanie Domenech; Bernal, Guillermo

    2011-02-01

    This article summarizes the definitions, means, and research of adapting psychotherapy to clients' cultural backgrounds. We begin by reviewing the prevailing definitions of cultural adaptation and providing a clinical example. We present an original meta-analysis of 65 experimental and quasi-experimental studies involving 8,620 participants. The omnibus effect size of d = .46 indicates that treatments specifically adapted for clients of color were moderately more effective with that clientele than traditional treatments. The most effective treatments tended to be those with greater numbers of cultural adaptations. Mental health services targeted to a specific cultural group were several times more effective than those provided to clients from a variety of cultural backgrounds. We recommend a series of research-supported therapeutic practices that account for clients' culture, with culture-specific treatments being more effective than generally culture-sensitive treatments. © 2010 Wiley Periodicals, Inc.

  13. Pilot-scale continuous recycling of growth medium for the mass culture of a halotolerant Tetraselmis sp. in raceway ponds under increasing salinity: a novel protocol for commercial microalgal biomass production.

    Science.gov (United States)

    Fon Sing, S; Isdepsky, A; Borowitzka, M A; Lewis, D M

    2014-06-01

    The opportunity to recycle microalgal culture medium for further cultivation is often hampered by salinity increases from evaporation and fouling by dissolved and particulate matter. In this study, the impact of culture re-use after electro-flocculation of seawater-based medium on growth and biomass productivity of the halotolerant green algal strain Tetraselmis sp., MUR 233, was investigated in pilot-scale open raceway ponds over 5months. Despite a salinity increase from 5.5% to 12% (w/v) NaCl, Tetraselmis MUR 233 grown on naturally DOC-enriched recycled medium produced 48-160% more ash free dry weight (AFDW) biomass daily per unit pond area than when grown on non-recycled medium. A peak productivity of 37.5±3.1gAFDWm(-2)d(-1) was reached in the recycled medium upon transition from ∼14% to ∼7% NaCl. The combination of high biomass-yielding mixotrophic growth under high salinity has been proven to be a successful sustainable cultivation strategy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Comparison of two commercial embryo culture media (SAGE-1 step single medium vs. G1-PLUSTM/G2-PLUSTM sequential media): Influence on in vitro fertilization outcomes and human embryo quality.

    Science.gov (United States)

    López-Pelayo, Iratxe; Gutiérrez-Romero, Javier María; Armada, Ana Isabel Mangano; Calero-Ruiz, María Mercedes; Acevedo-Yagüe, Pablo Javier Moreno de

    2018-04-26

    To compare embryo quality, fertilization, implantation, miscarriage and clinical pregnancy rates for embryos cultured in two different commercial culture media until D-2 or D-3. In this retrospective study, we analyzed 189 cycles performed in 2016. Metaphase II oocytes were microinjected and allocated into single medium (SAGE 1-STEP, Origio) until transferred, frozen or discarded; or, if sequential media were used, the oocytes were cultured in G1-PLUSTM (Vitrolife) up to D-2 or D-3 and in G2-PLUSTM (Vitrolife) to transfer. On the following day, the oocytes were checked for normal fertilization and on D-2 and D-3 for morphological classification. Statistical analysis was performed using the chi-square and Mann-Whitney tests in PASW Statistics 18.0. The fertilization rates were 70.07% for single and 69.11% for sequential media (p=0.736). The mean number of embryos with high morphological quality (class A/B) was higher in the single medium than in the sequential media: D-2 [class A (190 vs. 107, pcultured in single medium were frozen: 197 (21.00%) vs. sequential: 102 (11.00%), pculture in single medium yields greater efficiency per cycle than in sequential media. Higher embryo quality and quantity were achieved, resulting in more frozen embryos. There were no differences in clinical pregnancy rates.

  15. Identification of candidate agents active against N. ceranae infection in honey bees: establishment of a medium throughput screening assay based on N. ceranae infected cultured cells.

    Science.gov (United States)

    Gisder, Sebastian; Genersch, Elke

    2015-01-01

    Many flowering plants in both natural ecosytems and agriculture are dependent on insect pollination for fruit set and seed production. Managed honey bees (Apis mellifera) and wild bees are key pollinators providing this indispensable eco- and agrosystem service. Like all other organisms, bees are attacked by numerous pathogens and parasites. Nosema apis is a honey bee pathogenic microsporidium which is widely distributed in honey bee populations without causing much harm. Its congener Nosema ceranae was originally described as pathogen of the Eastern honey bee (Apis cerana) but jumped host from A. cerana to A. mellifera about 20 years ago and spilled over from A. mellifera to Bombus spp. quite recently. N. ceranae is now considered a deadly emerging parasite of both Western honey bees and bumblebees. Hence, novel and sustainable treatment strategies against N. ceranae are urgently needed to protect honey and wild bees. We here present the development of an in vitro medium throughput screening assay for the identification of candidate agents active against N. ceranae infections. This novel assay is based on our recently developed cell culture model for N. ceranae and coupled with an RT-PCR-ELISA protocol for quantification of N. ceranae in infected cells. The assay has been adapted to the 96-well microplate format to allow automated analysis. Several substances with known (fumagillin) or presumed (surfactin) or no (paromomycin) activity against N. ceranae were tested as well as substances for which no data concerning N. ceranae inhibition existed. While fumagillin and two nitroimidazoles (metronidazole, tinidazole) totally inhibited N. ceranae proliferation, all other test substances were inactive. In summary, the assay proved suitable for substance screening and demonstrated the activity of two synthetic antibiotics against N. ceranae.

  16. Identification of candidate agents active against N. ceranae infection in honey bees: establishment of a medium throughput screening assay based on N. ceranae infected cultured cells.

    Directory of Open Access Journals (Sweden)

    Sebastian Gisder

    Full Text Available Many flowering plants in both natural ecosytems and agriculture are dependent on insect pollination for fruit set and seed production. Managed honey bees (Apis mellifera and wild bees are key pollinators providing this indispensable eco- and agrosystem service. Like all other organisms, bees are attacked by numerous pathogens and parasites. Nosema apis is a honey bee pathogenic microsporidium which is widely distributed in honey bee populations without causing much harm. Its congener Nosema ceranae was originally described as pathogen of the Eastern honey bee (Apis cerana but jumped host from A. cerana to A. mellifera about 20 years ago and spilled over from A. mellifera to Bombus spp. quite recently. N. ceranae is now considered a deadly emerging parasite of both Western honey bees and bumblebees. Hence, novel and sustainable treatment strategies against N. ceranae are urgently needed to protect honey and wild bees. We here present the development of an in vitro medium throughput screening assay for the identification of candidate agents active against N. ceranae infections. This novel assay is based on our recently developed cell culture model for N. ceranae and coupled with an RT-PCR-ELISA protocol for quantification of N. ceranae in infected cells. The assay has been adapted to the 96-well microplate format to allow automated analysis. Several substances with known (fumagillin or presumed (surfactin or no (paromomycin activity against N. ceranae were tested as well as substances for which no data concerning N. ceranae inhibition existed. While fumagillin and two nitroimidazoles (metronidazole, tinidazole totally inhibited N. ceranae proliferation, all other test substances were inactive. In summary, the assay proved suitable for substance screening and demonstrated the activity of two synthetic antibiotics against N. ceranae.

  17. Effective clinical-scale production of dendritic cell vaccines by monocyte elutriation directly in medium, subsequent culture in bags and final antigen loading using peptides or RNA transfection.

    Science.gov (United States)

    Erdmann, Michael; Dörrie, Jan; Schaft, Niels; Strasser, Erwin; Hendelmeier, Martin; Kämpgen, Eckhart; Schuler, Gerold; Schuler-Thurner, Beatrice

    2007-09-01

    Dendritic cell (DC) vaccination approaches are advancing fast into the clinic. The major obstacle for further improvement is the current lack of a simple functionally "closed" system to generate standardized monocyte-derived (mo) DC vaccines. Here, we significantly optimized the use of the Elutra counterflow elutriation system to enrich monocytic DC precursors by (1) developing an algorithm to avoid red blood cell debulking and associated monocyte loss before elutriation, and (2) by elutriation directly in culture medium rather than phosphate-buffered saline. Upon elutriation the bags containing the collected monocytes are simply transferred into the incubator to generate DC progeny as the final "open" washing step is no longer required. Elutriation resulted in significantly more (> or = 2-fold) and purer DC than the standard gradient centrifugation/adherence-based monocyte enrichment, whereas morphology, maturation markers, viability, migratory capacity, and T cell stimulatory capacity were identical. Subsequently, we compared RNA transfection, as this is an increasingly used approach to load DC with antigen. Elutra-derived and adherence-derived DC could be electroporated with similar, high efficiency (on average >85% green fluorescence protein positive), and appeared also equal in antigen expression kinetics. Both Elutra-derived and adherence-derived DC, when loaded with the MelanA peptide or electroporated with MelanA RNA, showed a high T cell stimulation capacity, that is, priming of MelanA-specific CD8+ T cells. Our optimized Elutra-based procedure is straightforward, clearly superior to the standard gradient centrifugation/plastic adherence protocol, and now allows the generation of large numbers of peptide-loaded or RNA-transfected DC in a functionally closed system.

  18. Investigation of solid phase sorbents for the pre- concentration of pads from aqueous medium and their quantitation by high performance liquid chromatography-UV detection

    International Nuclear Information System (INIS)

    Waqar, F.; Jan, S.; Muhammad, B.; Ahmad, S.; Riaz, M.; Akram, N.

    2005-01-01

    A solid phase extraction method was optimized for the pre-concentration of polyaromatic hydrocarbons (PAHs) in water samples. Graphite powder and Lab scale locally synthesized styrene divinylbenzene (SDVB) Copolymer were used as sorbents for the extraction of PAHs and compared with commercially used C18 solid phase extraction cartridge (SPE). Various parameters were optimized to evaluate the extraction efficiencies, the best results were obtained by proper conditioning of extraction cartridges and desorption with suitable solvent. Percentage recoveries were enhanced by rinsing the sample bottles with acetonitrile and combining the rinse with the sample extract. Quantitative analysis was performed by High performance Liquid chromatography (HPLC) with UV detection. Many other parameters, including optimization of mobile phase, selection of HPLC Columns, sample-loading flow rate on extraction cartridge and weight of sorbent were performed to get optimal results. Percent recoveries obtained with synthesized copolymer were comparable with commercial cartridge, while graphite powder showed excellent retention but very poor recoveries. Obtained recoveries of selected PAHs were ranged from 80-87% with relative standard deviation <6%. Developed method was applied for the analysis of drinking water samples(author)

  19. Mixing Construction, Demolition and Excavation Waste and Solid Waste Compost for the Derivation of a Planting Medium for Use in the Rehabilitation of Quarries

    Science.gov (United States)

    Assaf, Eleni

    2015-04-01

    Lebanon's very high population density has been increasing since the end of the civil war in the early 1990s reaching 416.36 people per square kilometer. Furthermore, the influx of refugees from conflicts in the region has increased the resident population significantly. All these are exerting pressure on the country's natural resources, pushing the Lebanese to convert more forest and agricultural land into roads, buildings and houses. This has led to a building boom and rapid urbanization which in turn has created a demand for construction material - mainly rock, gravel, sand, etc. nearly all of which are locally acquired through quarrying to the tune of three million cubic meters annually. This boom has been interrupted by a war with Israel in 2006 which resulted in thousands of tonnes of debris. The increase in population has also led to an increase in solid waste generation with 1.57 million tonnes of solid waste generated in Lebanon per year. The combination of construction, demolition and excavation (CDE) waste along with the increase in solid waste generation has put a major stress on the country and on the management of its solid waste. Compounding this problem are the issues of quarries closure and rehabilitation and a decrease in forest and vegetative cover. The on-going research reported in this paper aims to provide an integrated solution to the stated problem by developing a "soil mix" derived from a mélange of the organic matter of the solid waste (compost), the CDE waste, and soil. Excavation and construction debris were ground to several sizes and mixed with compost and soil at different ratios. Replicates of these mixes and a set of control (regular soil) were used. In this mix, native and indicator plants are planted (in pots) from which the most productive mix will be selected for further testing at field level in later experiments. The plant species used are Mathiolla crassifolia, a native Lebanese plant and Zea mays (Corn), which is commonly

  20. Zastosowanie peptonu Peptobak-Bacutil tu hodowli merystematycznej tkanki Cymbidium Sw. [Application of the peptone Peptobak-Bacutil medium in the culture of meristem-tissue of Cymbidium Sw.

    Directory of Open Access Journals (Sweden)

    K. Kukułczanka

    2015-06-01

    Full Text Available In was found that peptone of Peptobak-Bacutil brand (produced in Poland fulfills perfectly the requirements in cultures in vitro of the meristem tissues of Cymbidium in the modified Tsuchiya's medium in a concentration of 2 g per 1. A decrease in the concentration to 1 g per 1. affects the growth intensity of the tissue, but an increase of concentration up to 3 g per 1. retards the transformation of protocorms into the plant-stage. Growth of the meristem tissue of Cymbidium in the medium consolidated with agar is weaker and the number of protocorms produced 2,3 times lower than in liquid media.

  1. Pulp fruit added to culture medium for in vitro orchid developmentPolpa de frutos adicionada ao meio de cultivo no crescimento in vitro de orquídea

    Directory of Open Access Journals (Sweden)

    Gilberto Rostirolla Batista de Souza

    2013-06-01

    Full Text Available As an additive in in vitro culture media, fruits have a great potential for facilitating economical orchid production because of lower technology requirements and the ease of obtaining raw materials to formulate culture media. We studied the in vitro growth of Cattleya bicolor Lindl. grown in a simplified culture medium supplemented with different kinds of fruit pulp. The experimental design was completely randomised, with eight seedlings per replication and ten replications per treatment, for a total of 80 seedlings per treatment. The culture medium was made using 150 g L -1 of pulp (without peel or seed from the following fruits: ripe Santa Cruz tomatoes (Solanum lycopersicum L., dwarf bananas (Musa cavendishii L. of intermediate ripeness, light green chayote (Sechium edule (Jacq. Sw, ripe papaya (Carica papaya L. or green coconut (Cocos nucifera L..The treatment control was MS 50 %. The treatments and the control were kept in a growth chamber for seven months before evaluating seedling survival percentage, shoot height, number of leaves, rooting percentage, root number, root length and dry masses of shoot and roots. The highest percentages of seedling survival were obtained using MS 50 %, banana and coconut medium. The seedling survival and rooting percentages illustrate that it is possible to emphasise the culture medium MS 50% and the culture medium supplemented with coconut on the most traditional culture medium with banana or tomato pulp. For the in vitro development of Cattleya bicolor Lindl., a simplified culture medium supplemented with coconut pulp is the most suitable for use as an alternative to MS 50%. A simplified culture medium supplemented with papaya pulp is not recommended for the in vitro development of Cattleya bicolor Lindl. Os frutos apresentam potencial para serem utilizados na elaboração de meios de cultivo para facilitar a produção de orquídeas em pequenas propriedades, contribuindo para a rentabilidade do cultivo

  2. Decolorization of the azo dye Acid Orange 51 by laccase produced in solid culture of a newly isolated Trametes trogii strain

    OpenAIRE

    Daâssi, Dalel; Zouari-Mechichi, Héla; Frikha, Fakher; Martínez, María Jesús; Nasri, M.; Mechichi, Tahar

    2013-01-01

    This study concerns the decolorization and detoxification of the azo dye Acid Orange 51 (AO51) by crude laccase from Trametes trogii produced in solid culture using sawdust as support media. A three-level Box?Behnken factorial design with four factors (enzyme concentration, 1-hydroxybenzotriazole (HBT) concentration, dye concentration and reaction time) combined with response surface methodology was applied to optimize AO51 decolorization. A mathematical model was developed showing the effect...

  3. Solid state protonic conductors II for fuel cells and sensors. Proceedings of the European workshop on solid state materials for low to medium temperature fuel cells and monitors, with special emphasis on proton conductors

    Energy Technology Data Exchange (ETDEWEB)

    Goodenough, J B; Jensen, J; Kleitz, M [eds.

    1983-01-01

    Solid electrolytes for chemical sensing, energy storage and conversion have been actively researched and developed since the early sixties. The zirconia fuel-cell electrolyser, the sodium-sulphur rechargeable battery, the oxygen sensor and lithium batteries can all be cited as significant developments from the field. Although of great potential the solid protonic conductors have somehow been ignored by comparison to the great interest that has been shown in, e.g., the lithium conductors. The long absence of any good, stable protonic conductors could easily explain this. The presence of water in the protonic conductors eliminates the possibility of high-temperature preparation and hence of conventional ceramic processing. Since solid electrolytes are used as dense ceramic membranes, difficulties with the fabrication of protonic electrilytes has been a strong disincentive. However, techniques have been developed for fabricating dense composite membranes; these contain free, but immobilized water that is lost at relatively low temperatures. Framework hydrates hold their water to higher temperatures. Although low-temperature ion-exchange preparations are possible, they yield weak ceramics. Nevertheless, their support on strong substrates, as reported in this conference, may provide an alternate way forward. A second workshop was organised on this theme at Hindsgavl Castle, Denmark, 1982. The aim was to compare the progress made in laboratories in Denmark, France and U.K. and also to review present and and future applications of fuel cells in a broader sense. Thirty scientists and representatives from the Commission of the European Communities, European Space Agency and the Daish Ministry of Energy participated. The proceedings cover all the papers of the workshop and the main comments and suggestions proposed during the discussions.

  4. Effects of solid-medium type on routine identification of bacterial isolates by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Anderson, Neil W; Buchan, Blake W; Riebe, Katherine M; Parsons, Lauren N; Gnacinski, Stacy; Ledeboer, Nathan A

    2012-03-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a rapid method for the identification of bacteria. Factors that may alter protein profiles, including growth conditions and presence of exogenous substances, could hinder identification. Bacterial isolates identified by conventional methods were grown on various media and identified using the MALDI Biotyper (Bruker Daltonics, Billerica, MA) using a direct smear method and an acid extraction method. Specimens included 23 Pseudomonas isolates grown on blood agar, Pseudocel (CET), and MacConkey agar (MAC); 20 Staphylococcus isolates grown on blood agar, colistin-nalidixic acid agar (CNA), and mannitol salt agar (MSA); and 25 enteric isolates grown on blood agar, xylose lysine deoxycholate agar (XLD), Hektoen enteric agar (HE), salmonella-shigella agar (SS), and MAC. For Pseudomonas spp., the identification rate to genus using the direct method was 83% from blood, 78% from MAC, and 94% from CET. For Staphylococcus isolates, the identification rate to genus using the direct method was 95% from blood, 75% from CNA, and 95% from MSA. For enteric isolates, the identification rate to genus using the direct method was 100% from blood, 100% from MAC, 100% from XLD, 92% from HE, and 87% from SS. Extraction enhanced identification rates. The direct method of MALDI-TOF analysis of bacteria from selective and differential media yields identifications of varied confidence. Notably, Staphylococci spp. from CNA exhibit low identification rates. Extraction enhances identification rates and is recommended for colonies from this medium.

  5. [Amylase production by Aureobasidium pullulans in liquid and solid media].

    Science.gov (United States)

    Lodato, P B; Forchiassin, F; Segovia de Huergo, M B

    1997-01-01

    Amylase production by a strain of Aureobasidium pullulans isolated in the laboratory was evaluated in liquid media (complex and synthetic) and in solid medium (wheat bran). There was an inhibitory effect in amylase production or amylase secretion by glucose. Asparagine was the best nitrogen source for amylase production (4-6 g/l). Only chlamidospores and melanin but not, amylase activity, were obtained with ammonium sulfate. Amylase production in solid culture was higher than the production obtained in the liquid media assayed. Optimum initial moisture content in solid culture ranged between 57 and 74%. No difference was observed in amylase production between solid media inoculated with cells grown in liquid or solid media.

  6. Seven years of experience in storage of solid radioactive wastes of low and medium activity on the surface or in concrete ditches

    International Nuclear Information System (INIS)

    Bardet, G.

    This experience in the transport and storage of radioactive waste leads to several reflections pertinent to large-scale nuclear installations throughout the world. The experience demonstrates that products of this kind can be centralized and confined by relatively simple processes which are safe and of reasonable cost. The necessary liaison between the producer of radioactive wastes and the group which takes charge of storage is emphasized for the choice of the waste conditioning procedure. It is a consistent part of the experience that the initial conditioning determines in large measure the handling and storage techniques. It is certain that the absence of a solution for long-term storage of radioactive wastes will lead the producers to consider and carry out conditioning which, although valuable, does not have all the same advantages for final disposition of the product. In the field of transport, the volumes and weights are important. It is desirable that a permanent storage center be near a railroad. It is stated that storage can be ensured under safe conditions and with acceptable costs of all solid wastes of low and average activity except those having more than a certain amount of alpha activity. For the latter products a solution can be found which is safe

  7. PdRu alloy nanoparticles of solid solution in atomic scale: outperformance towards formic acid electro-oxidation in acidic medium

    International Nuclear Information System (INIS)

    Miao, Kanghua; Luo, Yun; Zou, Jiasui; Yang, Jun; Zhang, Fengqi; Huang, Lin; Huang, Jie; Kang, Xiongwu; Chen, Shaowei

    2017-01-01

    Developing catalyst of high performance and low cost toward the electro-oxidation of formic acid on the anode of fuel cell is critical for the commercialization of direct formic acid fuel cells. Here we reported the synthesis of Pd x Ru 10-x (x = 1,3,5,7,9) nanoparticles (NPs) by concurrent reduction of Pd 2+ and Ru 2+ in polyol solution at 200 °C. The particle size of the obtained NPs was confined at 5–15 nm in diameter. X-ray diffraction (XRD) analysis revealed face-centered cubic (fcc) crystal structure for Pd x Ru 10-x (x = 3,5,7,9), with the lattice parameter proportional to the Pd content. The formation of the solid solution in atomic scale was confirmed for the alloy nanoparticles by XRD and the elemental mapping. Williamson-Hall method revealed that the stacking fault was dependent on the alloying extent of the alloy nanoparticles and reached the minimum for Pd 5 Ru 5 , which exhibited the highest activity towards formic acid oxidation among all these prepared samples, with mass activity of 12.6 times higher than that of commercial Pd/C. It was observed that the highest catalytic activity was in agreement with the minimum of the stacking fault of the alloy nanoparticles.

  8. Solid phase extraction of cadmium on 2-mercaptobenzothiazole loaded on sulfur powder in the medium of ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate and cold vapor generation-atomic absorption spectrometric determination

    International Nuclear Information System (INIS)

    Pourreza, N.; Ghanemi, K.

    2010-01-01

    A novel solid phase extractor for preconcentration of cadmium at ng L -1 levels has been developed. Cadmium ions were retained on a column packed with sulfur powder modified with 2-mercaptobenzothiazole (2-MBT) in the medium of 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim] + PF 6 - ) ionic liquid. The presence of ionic liquid during modification of sulfur enhanced the retention of cadmium ions on the column. The retained cadmium ions were eluted with 2 mol L -1 solution of HCl and measured by cold vapor generation-atomic absorption spectrometry (CVG-AAS). By using reaction cell-gas liquid separator (RC-GLS), gaseous cadmium vapors were produced and reached the atomic absorption spectrometer, instantaneously. The influence of different variables on both processes of solid phase extraction and CVG-AAS determination of cadmium ions was investigated. The calibration curve was linear in the range of 10-200 ng L -1 of cadmium in the initial solution with r = 0.9992 (n = 8) under optimum conditions. The limit of detection based on three times the standard deviation of the blank (3S b , n = 10) was 4.6 ng L -1 . The relative standard deviation (R.S.D.) of 25 and 150 ng L -1 of cadmium was 4.1 and 2.2% (n = 8), respectively. The procedure was validated by the analysis of a certified reference material (DORM-3), water and fish samples.

  9. Use of a solid-phase 3H-radioimmunoassay for the measurement of immunoglobulin produced in short-term cultures of antibody-secreting cells

    International Nuclear Information System (INIS)

    Mongini, P.K.A.; Heber-Katz, E.

    1982-01-01

    A solid-phase radioimmunoassay (RIA) for assaying immunoglobulin produced from antibody-secreting myeloma, hybridoma and immune spleen cells is described. Specific antibody is detected by culturing antibody-producing cells on antigen-coated flexible polyvinylchloride microtiter wells, washing away the cells, and measuring the bound specific antibody with tritiated affinity-purified anti-isotype reagents. Antibody produced from 10 3 myeloma cells can be detected with as little as 4 h of incubation. With 24-48 h of incubation, antibody from as few as approximately 3-15 myeloma, hybridoma or immune spleen plaque-forming cells (PFC) can be detected. This culture-well RIA has certain distinct advantages over the hemolytic PFC assay and RIA assays in which antibody in culture supernatants is measured. (Auth.)

  10. Development of an irrigation control device based on solar radiation and its adaptability for cultivation of high soluble solid tomato fruit in root zone restriction culture

    International Nuclear Information System (INIS)

    Nitta, M.; Shibuya, K.; Kubai, K.; Komatsu, H.; Hosokawa, T.; Nakamura, K.

    2009-01-01

    An irrigation control device based on solar radiation was developed to allow automatic irrigation management for high soluble solid tomato fruit production in root zone restriction culture. Its adaptability for long-term cultivation (planting carried out in early September and harvesting ending in late June) of high soluble solid tomato fruit in root zone restriction culture was examined. The following results were obtained: 1. The control device was composed of generally available electronic parts. A change of setting was possible for the irrigation starting point, the irrigation time period, and the once amount of irrigation. For the first irrigation of the day, one of two irrigation control modes can be chosen; the first determines irrigation dependent on the solar radiation after the irrigated time of the previous day. The second mode irrigates at a set time. 2. The correlation between the total integrated solar radiation and the evapotranspiration rate of tomato plants were investigated. Positive correlations were observed for each month from October to June. Moreover, total integrated solar radiation per unit evapotranspiration was different for each month. 3. In long-term cultivation of tomato fruit using this device, the marketable yield of high soluble solid tomato fruit (more than Brix 8%) was 9.7t/10a. 4. This device exhibited the necessary adaptability for use in long-term cultivation of high soluble solid tomato fruit in root zone restriction culture, by changing the set value of the irrigation starting point and the irrigation time period in accordance with the growth period

  11. Comparison of the rate of uptake and biologic effects of retinol added to human keratinocytes either directly to the culture medium or bound to serum retinol-binding protein

    International Nuclear Information System (INIS)

    Hodam, J.R.; St Hilaire, P.; Creek, K.E.

    1991-01-01

    Retinol circulates in the plasma bound to retinol-binding protein (RBP), but the mechanism by which retinol is transferred from RBP to target cells is not known. To study retinol delivery, human keratinocytes (HKc) were incubated with [3H]retinol added directly to the culture medium or bound to RBP and the uptake of [3H]retinol was determined at various times. During the first hour of incubation, the rate of [3H]retinol accumulation by HKc was about 40 times greater when the vitamin was added directly to the media rather than bound to RBP. Although maximal uptake of [3H]retinol added directly to the culture medium occurred at 3 h, the uptake of [3H]retinol from RBP was linear with time for at least 72 h. By 57 h, cell-associated [3H]retinol was the same whether it was added directly to the culture medium or bound to RBP. Excess unlabeled retinol or pretreatment of HKc with retinol had no effect on the uptake of [3H]retinol added directly to the culture medium or bound to RBP. Apo- but not holo-RBP was capable of competing with HKc for the uptake of [3H]retinol from RBP. No specific or saturable binding of 125I-labeled RBP to HKc cultured in the absence or the presence of retinol was found. The dose response of retinol inhibition of cholesterol sulfate synthesis and phorbol ester-induced ornithine decarboxylase activity or retinol modulation of keratin expression was the same whether the retinol was delivered to HKc bound to RBP or added directly to the medium. Our data support a mechanism for retinol delivery from RBP to HKc that does not involve cell-surface RBP receptors but instead suggest that the vitamin is first slowly released from RBP and then becomes cell-associated from the aqueous phase. This mechanism is consistent with the finding that HKc respond identically to retinol whether or not it is delivered to them bound to RBP

  12. Bio-detoxification of phorbol esters and other anti-nutrients of Jatropha curcas seed cake by fungal cultures using solid-state fermentation.

    Science.gov (United States)

    Sharath, B S; Mohankumar, B V; Somashekar, D

    2014-03-01

    Jatropha seed cake, a byproduct after biodiesel extraction, has several anti-nutrients and toxins. Solid-state fermentation was carried out for the detoxification of the Jatropha seed cake (JSC) using different fungal cultures. The reduction in the anti-nutritional components such as tannins, phytates, saponins, lectin and protease inhibitor, and phorbol esters on 6th, 9th, and 12th day of fermentation was analyzed. The phorbol ester content in the unfermented JSC was 0.83 mg/g, and the maximum degradation of phorbol esters to the extent of 75% was observed in the case of JSC fermented with Cunninghamella echinulata CJS-90. The phytate degradation in the fermented JSC was in the range of 65-96%. There was a gradual reduction of saponin content in the JSC from 6th to 12th day, and the reduction of saponin was in the range of 55-99% after solid-state fermentation. The trypsin inhibitor activity and lectin were 1,680 trypsin inhibitor units (TIU) per gram and 0.32 hemagglutinating unit in the unfermented JSC, respectively. Trypsin inhibitor activity and lectin could not be detected in JSC after 12th day of solid-state fermentation. Tannins accounted for 0.53% in unfermented JSC, and there was a marginal increase of tannins after solid-state fermentation. The results indicate that biological detoxification could be a promising method to reduce anti-nutritional compounds and toxins in the JSC.

  13. Psoralen production in hairy roots and adventitious roots cultures of Psoralea coryfolia.

    Science.gov (United States)

    Baskaran, P; Jayabalan, N

    2009-07-01

    Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension cultures or on agar solidified medium. Psoralen content was approximately 3 mg g(-1) DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious roots.

  14. Adaptation of a Commonly Used, Chemically Defined Medium for Human Embryonic Stem Cells to Stable Isotope Labeling with Amino Acids in Cell Culture

    DEFF Research Database (Denmark)

    Liberski, A. R.; Al-Noubi, M. N.; Rahman, Z. H.

    2013-01-01

    Metabolic labeling with stable isotopes is a prominent technique for comparative quantitative proteomics, and stable isotope labeling with amino acids in cell culture (SILAC) is the most commonly used approach. SILAC is, however, traditionally limited to simple tissue culture regimens and only ra...

  15. Methodology for monitoring gold nanoparticles and dissolved gold species in culture medium and cells used for nanotoxicity tests by liquid chromatography hyphenated to inductively coupled plasma-mass spectrometry.

    Science.gov (United States)

    López-Sanz, Sara; Fariñas, Nuria Rodríguez; Vargas, Rosario Serrano; Martín-Doimeadios, Rosa Del Carmen Rodríguez; Ríos, Ángel

    2017-03-01

    An analytical methodology based on coupling reversed-phase liquid chromatography (HPLC) to an inductively coupled plasma mass spectrometry (ICP-MS) has been developed for the characterization and identification of gold nanoparticles (AuNPs) and gold dissolved species (Au 3+ ) in culture medium (Dulbecco's Modified Eagle Medium, DMEM) and HeLa cells (a human cervical adenocarcinoma cell line) used in nanotoxicity tests. The influence of the culture medium was also studied and the method applied for nanotoxicity tests. It was also observed that AuNPs can undergo an oxidation process in the supernatants and only a small amount of AuNPs and dissolved Au 3+ was associated with cells. To evaluate the biological impact of AuNPs, a classical viability assay onto HeLa cells was performed using cellular media DMEM in the presence of increasing dosage of 10nm AuNPs. The results showed that 10nm AuNPs exhibit a slight toxic effect. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Long-term storage of clinical samples in CyMol® medium for PNA- FISH® and culturing from the eSwab™ system

    DEFF Research Database (Denmark)

    Larsen, Lone Heimann; Xu, Yijuan; Pedersen, Malene Schibler

    Objectives: A steadily growing diversity of bacteria is reported in foreign body infections, and culture-independent methods have been shown to supplement established culture methods. Therefore, sampling and preservation of specimens have become an important issue. We report here experience from...... analytical methods. Methods: Sampling for both culture-dependent and -independent analyses were done over a period of two years. Specimens were transferred directly to the lab, and cultures of tissue biopsies, joint fluid, sonication fluid from the prosthesis components, and eSwab™ (Copan, Italy) were...... performed within 24 h after sampling. The corresponding specimens for culture-independent methods were stored at -80°C until analyzed in batchs. Specimens for fluorescence in situ hybridization (FISH) analysis were stored for app. one year at -80°C in CyMol® (Copan, Italy), an alcohol based media, before...

  17. Production of medium-chain fatty acids and higher alcohols by a synthetic co-culture grown on carbon monoxide or syngas

    NARCIS (Netherlands)

    Diender, M.; Stams, A.J.M.; Machado de Sousa, D.Z.

    2016-01-01

    Background
    Synthesis gas, a mixture of CO, H2, and CO2, is a promising renewable feedstock for bio-based production of organic chemicals. Production of medium-chain fatty acids can be performed via chain elongation, utilizing acetate and ethanol as main substrates. Acetate and ethanol are main

  18. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  19. Micropropagation of Alstroemeria in liquid medium using slow release of medium components

    NARCIS (Netherlands)

    Klerk, de G.J.M.; Brugge, ter J.

    2010-01-01

    Alstroemeria rhizomes were micropropagated on semi-solid medium (AM) and in liquid medium (LM). In LM, growth was much enhanced (ca. 70%). Adequate gas exchange was crucial. This was obtained by agitation and in static medium by a sufficient large contact area of the explant and the gaseous

  20. Optimization of culture medium for anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl for microbial enhanced oil recovery.

    Science.gov (United States)

    Zhao, F; Mandlaa, M; Hao, J; Liang, X; Shi, R; Han, S; Zhang, Y

    2014-08-01

    Response surface methodology was employed to enhance the anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl. Glycerol is a promising carbon source used to anaerobically produce rhamnolipid. In a Plackett-Burman design, glycerol, KH2 PO4 and yeast extract were significant factors. The proposed optimized medium contained the following: 46·55 g l(-1) glycerol; 3 g l(-1) NaNO3 ; 5·25 g l(-1) K2 HPO4 ·3H2 O; 5·71 g l(-1) KH2 PO4 ; 0·40 g l(-1) MgSO4 ·7H2 O; 0·13 g l(-1) CaCl2 ; 1·0 g l(-1) KCl; 1·0 g l(-1) NaCl; and 2·69 g l(-1) yeast extract. Using this optimized medium, we obtained an anaerobic yield of rhamnolipid of 3·12 ± 0·11 g l(-1) with a 0·85-fold increase. Core flooding test results also revealed that Ps. stutzeri Rhl grown in an optimized medium enhanced the oil recovery efficiency by 15·7%, which was 6·6% higher than in the initial medium. Results suggested that the optimized medium is a promising nutrient source that could effectively mobilize oil by enhancing the in situ production of rhamnolipid. The ex situ application of rhamnolipid for microbial enhanced oil recovery (MEOR) is costly and complex in terms of rhamnolipid production, purification and transportation. Compared with ex situ applications, the in situ production of rhamnolipid in anaerobic oil reservoir is more advantageous for MEOR. This study is the first to report the anaerobic production optimization of rhamnolipid. Results showed that the optimized medium enhanced not only the anaerobic production of rhamnolipid but also crude oil recovery. © 2014 The Society for Applied Microbiology.

  1. Changes in the metabolic footprint of placental explant-conditioned medium cultured in different oxygen tensions from placentas of small for gestational age and normal pregnancies.

    LENUS (Irish Health Repository)

    Horgan, R P

    2012-01-31

    Being born small for gestational age (SGA) confers significantly increased risks of perinatal morbidity and mortality. Accumulating evidence suggests that an SGA fetus results from a poorly perfused and abnormally developed placenta. Some of the placental features seen in SGA, such as abnormal cell turnover and impaired nutrient transport, can be reproduced by culture of placental explants in hypoxic conditions. Metabolic footprinting offers a hypothesis-generating strategy to investigate factors absorbed by and released from this tissue in vitro. Previously, metabolic footprinting of the conditioned culture media has identified differences in placental explants cultured under normoxic and hypoxic conditions and between normal pregnancies and those complicated by pre-eclampsia. In this study we aimed to examine the differences in the metabolic footprint of placental villous explants cultured at different oxygen (O(2)) tensions between women who deliver an SGA baby (n = 9) and those from normal controls (n = 8). Placental villous explants from cases and controls were cultured for 96 h in 1% (hypoxic), 6% (normoxic) and 20% (hyperoxic) O(2). Metabolic footprints were analysed by Ultra Performance Liquid Chromatography coupled to an electrospray hybrid LTQ-Orbitrap Mass Spectrometry (UPLC-MS). 574 metabolite features showed significant difference between SGA and normal at one or more of the oxygen tensions. SGA explant media cultured under hypoxic conditions was observed, on a univariate level, to exhibit the same metabolic signature as controls cultured under normoxic conditions in 49% of the metabolites of interest, suggesting that SGA tissue is acclimatised to hypoxic conditions in vivo. No such behaviour was observed under hyperoxic culture conditions. Glycerophospholipid and tryptophan metabolism were highlighted as areas of particular interest.

  2. Drug and radiation sensitivity measurements of successful primary monolayer culturing of human tumor cells using cell-adhesive matrix and supplemented medium

    International Nuclear Information System (INIS)

    Baker, F.L.; Spitzer, G.; Ajani, J.A.

    1986-01-01

    The limitations of the agar suspension culture method for primary culturing of human tumor cells prompted development of a monolayer system optimized for cell adhesion and growth. This method grew 83% of fresh human tumor cell biopsy specimens, cultured and not contaminated, from a heterogeneous group of 396 tumors including lung cancer (93 of 114, 82%); melanoma (54 of 72, 75%); sarcoma (46 of 59, 78%); breast cancer (35 of 39, 90%); ovarian cancer (16 of 21, 76%); and a miscellaneous group consisting of gastrointestinal, genitourinary, mesothelioma, and unknown primaries (78 of 91, 86%). Cell growth was characterized morphologically with Papanicolaoustained coverslip cultures and cytogenetically with Giemsastained metaphase spreads. Morphological features such as nuclear pleomorphism, chromatin condensation, basophilic cytoplasm, and melanin pigmentation were routinely seen. Aneuploid metaphases were seen in 90% of evaluable cultures, with 15 of 28 showing 70% or more aneuploid metaphases. Colony-forming efficiency ranged between 0.01 and 1% of viable tumor cells, with a median efficiency of 0.2%. This culture system uses a low inoculum of 25,000 viable cells per well which permitted chemosensitivity testing of nine drugs at four doses in duplicate from 2.2 X 10(6) viable tumor cells and radiation sensitivity testing at five doses in quadruplicate from 0.6 X 10(6) cells. Cultures were analyzed for survival by computerized image analysis of crystal violet-stained cells. Drug sensitivity studies showed variability in sensitivity and in survival curve shape with exponential cell killing for cisplatin, Adriamycin, and etoposide, and shouldered survival curves for 5-fluorouracil frequently seen. Radiation sensitivity studies also showed variability in both sensitivity and survival curve shape. Many cultures showed exponential cell killing, although others had shouldered survival curves

  3. Some major deviations for biomass determination by indirect method and estimation based on alkali consumption. [Ratio of cell mass produced and alkali consumed; diesel fuel culture medium

    Energy Technology Data Exchange (ETDEWEB)

    Concone, B R.V.; Doin, P A; Pinto, A G

    1978-01-01

    Some factors like the variation of the liquid volume, the variation of cellular nitrogen content and the mass of cells taken with the samples during batch cultivation of microorganisms on diesel oil, were considered for the computation of the ratio between cell mass produced and the mass of alkali consumed to maintain constant the pH of the fermentation medium. The results obtained showed that if such ratios are computed with cell concentration instead of cell mass the deviations can be of the order of 27% caused by the variation of the liquid medium volume. Otherwise, the results showed also that those ratios are variable during batch cultivation on diesel oil probably because of the variations on the nitrogen content of microorganisms. The relative difference between the mass of cells measured and the mass of cells calculated from the alkali consumption curve can be of the order of 63%.

  4. Effect Of Explant Source And Different Medium Culture On Friable Embryogenic Callus Induction Of Four Cultivars Of Cassava Manihot Esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Simplice Prosper Yandia

    2015-08-01

    Full Text Available In order to obtain Friable Embryogenic Callus FEC for protoplast isolation we have evaluated in this research the competance for Friable Embryogenic Callus FEC of four cassava cultivars M61033 Rendre Yalipe and Six-mois in media containing MS supplemented with 8mgl 24-D MS supplemented with 10 mgl BAP and GD supplemented with 12mgl picloram using apical bud AB and immature leaves lobes ILL as explants. In general in the medium GD12mgl picloram the highest efficiencies of FEC ranged from 58 to 87 and the highest score of FEC ranged from 4.2 to 5.4 with explants AB however we have observed with explants ILL the efficiencies of somatic embryos ranged form 41 to 75 and the score ranged from 4.1 to 4.4. The mediums MS28 mgl 24-D have induced with explants AB the efficiencies of FEC ranged from 43 to 57 and the score ranged from 3.1 to 3.8 however with ILL explants the efficiencies of FEC ranged from 39 to 49 and the score ranged from 2.9 to 3.7. The least FEC were observed in the medium MS210 mgl BAP with BA explants however the efficiencies ranged from 6 to 11 and the score ranged from 1.1 to 1.8. Whereas the efficiencies of FEC with ILL explants ranged from 4 to 7 and the score ranged from 0.5 to 0.8. All of four cultivars showed capability of producing FEC although their efficiency varied according to gonotype donors explants and medium taking into acount. Abbreviations GD Gressoff and Doy MS Murashige and Skoog 24-dichloro phenoxyacetic acid BAP Benzylamino-purin-Acid AB Apical Bud ILL Immature Leaves lobes

  5. Human umbilical cord Wharton's jelly stem cells undergo enhanced chondrogenic differentiation when grown on nanofibrous scaffolds and in a sequential two-stage culture medium environment.

    Science.gov (United States)

    Fong, Chui-Yee; Subramanian, Arjunan; Gauthaman, Kalamegam; Venugopal, Jayarama; Biswas, Arijit; Ramakrishna, Seeram; Bongso, Ariff

    2012-03-01

    The current treatments used for osteoarthritis from cartilage damage have their disadvantages of donor site morbidity, complicated surgical interventions and risks of infection and graft rejection. Recent advances in tissue engineering have offered much promise in cartilage repair but the best cell source and in vitro system have not as yet been optimised. Human bone marrow mesenchymal stem cells (hBMSCs) have thus far been the cell of choice. However, we derived a unique stem cell from the human umbilical cord Wharton's jelly (hWJSC) that has properties superior to hBMSCs in terms of ready availability, prolonged stemness characteristics in vitro, high proliferation rates, wide multipotency, non-tumorigenicity and tolerance in allogeneic transplantation. We observed enhanced cell attachment, cell proliferation and chondrogenesis of hWJSCs over hBMSCs when grown on PCL/Collagen nanoscaffolds in the presence of a two-stage sequential complex/chondrogenic medium for 21 days. Improvement of these three parameters were confirmed via inverted optics, field emission scanning electron microscopy (FESEM), MTT assay, pellet diameters, Alcian blue histology and staining, glycosaminglycans (GAG) and hyaluronic acid production and expression of key chondrogenic genes (SOX9, Collagen type II, COMP, FMOD) using immunohistochemistry and real-time polymerase chain reaction (qRT-PCR). In separate experiments we demonstrated that the 16 ng/ml of basic fibroblast growth factor (bFGF) present in the complex medium may have contributed to driving chondrogenesis. We conclude that hWJSCs are an attractive stem cell source for inducing chondrogenesis in vitro when grown on nanoscaffolds and exposed sequentially first to complex medium and then followed by chondrogenic medium.

  6. Bile culture

    Science.gov (United States)

    Culture - bile ... is placed in a special dish called a culture medium to see if bacteria, viruses, or fungi ... Chernecky CC, Berger BJ. Body fluid - anaerobic culture. In: ... . 6th ed. St Louis, MO: Elsevier Saunders; 2013:225-226. Kim AY, ...

  7. Toward a feline-optimized culture medium: impact of ions, carbohydrates, essential amino acids, vitamins, and serum on development and metabolism of in vitro fertilization-derived feline embryos relative to embryos grown in vivo.

    Science.gov (United States)

    Herrick, Jason R; Bond, Jennifer B; Magarey, Genevieve M; Bateman, Helen L; Krisher, Rebecca L; Dunford, Susan A; Swanson, William F

    2007-05-01

    The objective of this study was to define the physiologic needs of domestic cat embryos to facilitate development of a feline-specific culture medium. In a series of factorial experiments, in vivo-matured oocytes (n = 2040) from gonadotropin-treated domestic cats were inseminated in vitro to generate embryos (n = 1464) for culture. In the initial study, concentrations of NaCl (100.0 vs. 120.0 mM), KCl (4.0 vs. 8.0 mM), KH(2)PO(4) (0.25 vs. 1.0 mM), and the ratio of CaCl(2) to MgSO(4)-7H(2)O (1.0:2.0 mM vs. 2.0:1.0 mM) in the medium were evaluated during Days 1-6 (Day 0: oocyte recovery and in vitro fertilization [IVF]) of culture. Subsequent experiments assessed the effects of varying concentrations of carbohydrate (glucose, 1.5, 3.0, or 6.0 mM; l-lactate, 3.0, 6.0, or 12.0 mM; and pyruvate, 0.1 or 1.0 mM) and essential amino acids (EAAs; 0, 0.5, or 1.0x) in the medium during Days 1-3 and Days 3-6 of culture. Inclusion of vitamins (0 vs. 1.0x) and fetal calf serum (FCS; 0 vs. 5% [v/v]) in the medium also was evaluated during Days 3-6. Development and metabolism of IVF embryos on Day 3 or Day 6 were compared to age-matched in vivo embryos recovered from naturally mated queens. A feline-optimized culture medium (FOCM) was formulated based on these results (100.0 mM NaCl, 8.0 mM KCl, 1.0 mM KH(2)PO(4), 2.0 mM CaCl(2), 1.0 mM MgSO(4), 1.5 mM glucose, 6.0 mM L-lactate, 0.1 mM pyruvate, and 0x EAAs with 25.0 mM NaHCO(3), 1.0 mM alanyl-glutamine, 0.1 mM taurine, and 1.0x nonessential amino acids) with 0.4% (w/v) BSA from Days 0-3 and 5% FCS from Days 3-6. Using this medium, ~70% of cleaved embryos developed into blastocysts with profiles of carbohydrate metabolism similar to in vivo embryos. Our results suggest that feline embryos have stage-specific responses to carbohydrates and are sensitive to EAAs but are still reliant on one or more unidentified components of FCS for optimal blastocyst development.

  8. Optimizing culture medium composition to improve oligodendrocyte progenitor cell yields in vitro from subventricular zone-derived neural progenitor cell neurospheres.

    Directory of Open Access Journals (Sweden)

    Paula G Franco

    Full Text Available Neural Stem and Progenitor Cells (NSC/NPC are gathering tangible recognition for their uses in cell therapy and cell replacement therapies for human disease, as well as a model system to continue research on overall neural developmental processes in vitro. The Subventricular Zone is one of the largest NSC/NPC niches in the developing mammalian Central Nervous System, and persists through to adulthood. Oligodendrocyte progenitor cell (OPC enriched cultures are usefull tools for in vitro studies as well as for cell replacement therapies for treating demyelination diseases. We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield. The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation. Furthermore, heparin addition to the culture media contributed to further increase overall culture yields. The OPC generated by this protocol were able to mature into Myelin Basic Protein-expressing cells and to interact with neurons in an in vitro co-culture system. As a whole, we describe an optimized in vitro method for increasing OPC.

  9. Rapid Two-Step Procedure for Large-Scale Purification of Pediocin-Like Bacteriocins and Other Cationic Antimicrobial Peptides from Complex Culture Medium

    Science.gov (United States)

    Uteng, Marianne; Hauge, Håvard Hildeng; Brondz, Ilia; Nissen-Meyer, Jon; Fimland, Gunnar

    2002-01-01

    A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a low-pressure, reverse-phase column and the bacteriocins were detected as major optical density peaks upon elution with propanol. More than 80% of the activity that was initially in the culture supernatant was recovered in both purification steps, and the final bacteriocin preparation was more than 90% pure as judged by analytical reverse-phase chromatography and capillary electrophoresis. PMID:11823243

  10. Rapid Two-Step Procedure for Large-Scale Purification of Pediocin-Like Bacteriocins and Other Cationic Antimicrobial Peptides from Complex Culture Medium

    OpenAIRE

    Uteng, Marianne; Hauge, Håvard Hildeng; Brondz, Ilia; Nissen-Meyer, Jon; Fimland, Gunnar

    2002-01-01

    A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a lo...

  11. Effect of different surfactants in biorelevant medium on the secretion of a lipophilic compound in lipoproteins using Caco-2 cell culture

    DEFF Research Database (Denmark)

    Karpf, Ditte M; Holm, René; Garafalo, Carole

    2006-01-01

    The impact of a pharmaceutical relevant metabolizable, ionic surfactant or two synthetic, nonionic surfactants on the absorption and lipoprotein incorporation of a lipophilic drug, retinol, was studied in the Caco-2 cell culture. Filter-grown monolayers of Caco-2 cells were incubated for 20 h...

  12. English as a Medium of Instruction in East Asia's Higher Education Sector: A Critical Realist Cultural Political Economy Analysis of Underlying Logics

    Science.gov (United States)

    Kedzierski, Matt

    2016-01-01

    As discourses of globalisation and the knowledge-based economy become increasingly influential in both policy-making and in public debates about education, employability and national competitiveness--the choice of language in the classroom takes on a strategic importance. The paper employs a critical realist Cultural Political Economy lens to…

  13. Influence of culture medium pH on the production of CGTase by Bacillus firmus Strain No. 37 - doi: 10.4025/actascitechnol.v35i3.15882

    Directory of Open Access Journals (Sweden)

    Jéssica Bravin Carmello

    2013-06-01

    Full Text Available The enzyme cyclomaltodextrin-glucanotransferase (CGTase is a transglicosidase able to convert corn starch into cyclodextrin (CD. CDs are widely applied in industry given the ability to form inclusion complexes with a great variety of organic molecules. Regarding the optimum pH of CGTase, values reported in the literature vary according to the enzyme producing microorganism, being 8.0 the optimum pH of CGTase produced by Bacillus firmus Strain No. 37. This work studied the influence of the pH of culture medium with different concentration of nutrients on the production of the enzyme CGTase by Bacillus firmus Strain No. 37. For this purpose, the microorganism was grown in three culture media with different concentrations of carbon and nitrogen. The pH control was performed by adding sodium carbonate. The fermentation process was analyzed by the following methods: Bradford (1976 method to determine soluble proteins, DNS method to analyze sugars, and the method of complexation with β-CD to analyze the enzyme activity. The best result for CGTase enzyme activity was 0.22 U mL-1, obtained with medium containing 2.0% soluble corn starch and yeast extract, and pH 8.3.  

  14. Micropropagation of Achillea millefolium L. on half-strength ms medium and direct rooting and acclimatization of microshoots in hydroponic culture

    Directory of Open Access Journals (Sweden)

    Marković Marija

    2015-01-01

    Full Text Available The aim of this study was to determine the possibility of micropropagation of the medicinal plant A. millefolium on half-strength MS medium and ex vitro rooting and acclimatization of the obtained microshoots in hydroculture in order to establish an efficient production method. Two explant types were used: basal and terminal cuttings, and better results were achieved when terminal cuttings were used. The development of shoots in the multiplication phase was successful with a regeneration percentage of 100%. Ex vitro rooting in a modified Hoagland nutrient solution was successful (83%, but the percentage of in vitro rooting on half-strength MS medium without hormones was higher (95%. However, bearing in mind that mass production of A. millefolium is more efficient when the phase of in vitro rooting is excluded, this method could be recommended for commercial propagation of this medicinal plant. It is necessary to conduct additional research in order to optimize the composition, EC and pH value of the hydroponic nutrient solution. [Projekat Ministarstva nauke Republike Srbije, br. 43007

  15. The C2H2-type transcription factor, FlbC, is involved in the transcriptional regulation of Aspergillus oryzae glucoamylase and protease genes specifically expressed in solid-state culture.

    Science.gov (United States)

    Tanaka, Mizuki; Yoshimura, Midori; Ogawa,