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Sample records for culture medium increased

  1. 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.

  2. Mouse preimplantation embryo responses to culture medium osmolarity include increased expression of CCM2 and p38 MAPK activation

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    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.

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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)

  9. 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.

  10. 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.

  11. INNOVATIVE CULTURE IN SMALL AND MEDIUM ENTERPRISES

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    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.

  12. 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 ...

  13. 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 ...

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

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

    2015-01-01

    supernatant increased significantly after 1.5 min to 466 ± 543 pg·mL-1 (p led to a rapid disturbance of the HUVEC with stress fiber formation, disconnection of cell-cell contacts and an altered prostacyclin secretion, which had regressed nearly completely after 12 hours. Therefore, the evaluation of HUVEC on body foreign materials should be performed not earlier than 12 hours after cell culture medium exchange to avoid a misinterpretation of the endothelial cell morphological state. This procedure minimizes the risk of a misinterpretation of the endothelial cell morphology - caused by the culture medium exchange and not by the interaction between biomaterials and HUVEC.

  15. 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)

  16. 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.

  17. 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 ...

  18. 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...

  19. 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

  20. 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.

  1. 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.

  2. 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)

  3. Culture medium for amylase production by toxigenic fungi

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    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.

  4. 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

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

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    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.

  6. 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...

  7. 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.

  8. 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.

  9. Increasing Understanding of Cultural Differences

    Science.gov (United States)

    Creeden, Jack; Kelly-Aguirre, Eileen; Visser, Aric

    2016-01-01

    Many high school and university students return home from global programs and often report they have changed as a result of the experience. Global educators assume the act of participating in global education programs (such as high school study abroad) will open students' eyes to the complexities of another culture because students have been…

  10. 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 ...

  11. 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.

  12. 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.

  13. Increasing synthetic serum substitute (SSS) concentrations in P1 glucose/phosphate-free medium improves implantation rate: a comparative study.

    Science.gov (United States)

    Ben-Yosef, D; Yovel, I; Schwartz, T; Azem, F; Lessing, J B; Amit, A

    2001-11-01

    To assess the comparative efficacy of IVF medium (MediCult, with 5.2 mM glucose) and a glucose/phosphate-free medium, P1 (Irvine Scientific), and to investigate the influence of increasing the serum supplementation (synthetic serum substitute; SSS; Irvine Scientific) to P1 on embryo development and implantation. Patients were randomly assigned to IVF medium (Group 1, cycles n = 172) or P1 supplemented with 10% SSS (Group 2, cycles n = 229) according to the medium scheduled for use on the day of oocyte retrieval. Another 555 IVF consequent cycles (Group 3) were performed using increased SSS concentrations (20%) in P1 medium. In this large series of IVF cycles, we herein demonstrate that significantly higher pregnancy and implantation rates were found when embryos were cultured in glucose/phosphate-free medium P1 supplemented with 20% SSS compared to supplementation with the lower SSS concentration and with IVF medium.

  14. Harvesting Chlorella vulgaris by natural increase in pH: effect of medium composition.

    Science.gov (United States)

    Nguyen, Thi Dong Phuong; Frappart, Matthieu; Jaouen, Pascal; Pruvost, Jérémy; Bourseau, Patrick

    2014-01-01

    The freshwater microalga Chlorella vulgaris was harvested by autoflocculation resulting from the precipitation of magnesium or calcium compounds induced by a slow increase in pH in the absence of CO2 input. Autoflocculation was tested in two culture media with, respectively, ammonium (NH4+) and nitrate (NO3-) ions as nitrogen source. The culture pH increased because of photosynthesis and CO2 stripping. pH rose to 11 after 8 h in the NO3- medium, but did not exceed 9 in the NH4+ medium. No flocculation took place in any of the media. Autoflocculation tests were repeated in the NO(3-)-based culture medium by progressively increasing the concentrations of Ca2+ and Mg2+ until inorganic compounds precipitated and flocculated microalgae. The minimal concentrations for flocculation were found to be 120 mg Ca2 L(-1) and 1000 mg Mg2+ L(-1). These values were, respectively, 3.5 times and 20 times higher than those allowing flocculation by NaOH addition. Energy-dispersive X-ray spectroscopy, zeta potential measurement, and ionic chromatography suggest that the mechanisms involved are different. The rate of cell removal was close to 90% in both cases, but cells were more concentrated in the aggregates obtained by magnesium compound precipitation, with an estimated concentration close to 33 g (dry matter) L(-1), against 19 g L(-1) for calcium phosphates.

  15. [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.

  16. 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

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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)

  3. 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)

  4. 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

  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. 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...

  7. 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.

  8. 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.

  9. 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...

  10. 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

  11. 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.

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. Increased dipicolinic acid production with an enhanced spoVF operon in Bacillus subtilis and medium optimization.

    Science.gov (United States)

    Takahashi, Fumikazu; Sumitomo, Nobuyuki; Hagihara, Hiroshi; Ozaki, Katsuya

    2015-01-01

    Dipicolinic acid (DPA) is a multi-functional agent for cosmetics, antimicrobial products, detergents, and functional polymers. The aim of this study was to design a new method for producing DPA from renewable material. The Bacillus subtilis spoVF operon encodes enzymes for DPA synthase and the part of lysine biosynthetic pathway. However, DPA is only synthesized in the sporulation phase, so the productivity of DPA is low level. Here, we report that DPA synthase was expressed in vegetative cells, and DPA was produced in the culture medium by replacement of the spoVFA promoter with other highly expressed promoter in B. subtilis vegetative cells, such as spoVG promoter. DPA levels were increased in the culture medium of genetically modified strains. DPA productivity was significantly improved up to 29.14 g/L in 72 h culture by improving the medium composition using a two-step optimization technique with the Taguchi methodology.

  17. 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.

  18. 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...

  19. 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...

  20. 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...

  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. 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.

  3. 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.

  4. 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.

  5. 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...

  6. 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)

  7. 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.

  8. 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.

  9. 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

  10. Plant Explants Grown on Medium Supplemented with Fe3O4 Nanoparticles Have a Significant Increase in Embryogenesis

    Directory of Open Access Journals (Sweden)

    Inese Kokina

    2017-01-01

    Full Text Available Development of nanotechnology leads to the increasing release of nanoparticles in the environment that results in accumulation of different NPs in living organisms including plants. This can lead to serious changes in plant cultures which leads to genotoxicity. The aims of the present study were to detect if iron oxide NPs pass through the flax cell wall, to compare callus morphology, and to estimate the genotoxicity in Linum usitatissimum L. callus cultures induced by different concentrations of Fe3O4 nanoparticles. Two parallel experiments were performed: experiment A, where flax explants were grown on medium supplemented with 0.5 mg/l, 1 mg/l, and 1.5 mg/l Fe3O4 NPs for callus culture obtaining, and experiment B, where calluses obtained from basal MS medium were transported into medium supplemented with concentrations of NPs identical to experiment A. Obtained results demonstrate similarly in both experiments that 25 nm Fe3O4 NPs pass into callus cells and induce low toxicity level in the callus cultures. Nevertheless, calluses from experiment A showed 100% embryogenesis in comparison with experiment B where 100% rhizogenesis was noticed. It could be associated with different stress levels and adaptation time for explants and calluses that were transported into medium with Fe3O4 NPs supplementation.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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)

  16. 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)

  17. 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.

     

  18. 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.)

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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

  14. 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…

  15. 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...

  16. 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

  17. 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)

  18. Increase in the biomass of some green algae species in nitrate and ammonium mediums depending on auto-, mixo- or heterotrophic conditions

    Directory of Open Access Journals (Sweden)

    Stefan Gumiński

    2014-01-01

    Full Text Available The increase in total dry mass and protein in cultures of Chlorella pyrenoidosa, Scenedesmus quadricauda and Ankistrodesmus acicularis was studied. Under autotrophic conditions, increases in dry mass were, as a rule, larger in the nitrate medium than in the ammonium one, under mixotrophic conditions the situation was reversed and in the case of heterotrophy, the individual species reacted differently. The dependence ot the protein content increase on the nitrate or ammonium form of the medium was not clear. Changes in time of the pH and rH of the mediums were followed and the interdependence of these changes with the production of biomass is discussed.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. 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.

  7. 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

  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. 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

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. 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)

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. 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...

  1. 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).

  2. 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).

  3. 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.

  4. 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.

  5. Fungi outcompete bacteria under increased uranium concentration in culture media

    International Nuclear Information System (INIS)

    Mumtaz, Saqib; Streten-Joyce, Claire; Parry, David L.; McGuinness, Keith A.; Lu, Ping; Gibb, Karen S.

    2013-01-01

    As a key part of water management at the Ranger Uranium Mine (Northern Territory, Australia), stockpile (ore and waste) runoff water was applied to natural woodland on the mine lease in accordance with regulatory requirements. Consequently, the soil in these Land Application Areas (LAAs) presents a range of uranium concentrations. Soil samples were collected from LAAs with different concentrations of uranium and extracts were plated onto LB media containing no (0 ppm), low (3 ppm), medium (250 ppm), high (600 ppm) and very high (1500 ppm) uranium concentrations. These concentrations were similar to the range of measured uranium concentrations in the LAAs soils. Bacteria grew on all plates except for the very high uranium concentrations, where only fungi were recovered. Identifications based on bacterial 16S rRNA sequence analysis showed that the dominant cultivable bacteria belonged to the genus Bacillus. Members of the genera Paenibacillus, Lysinibacillus, Klebsiella, Microbacterium and Chryseobacterium were also isolated from the LAAs soil samples. Fungi were identified by sequence analysis of the intergenic spacer region, and members of the genera Aspergillus, Cryptococcus, Penicillium and Curvularia were dominant on plates with very high uranium concentrations. Members of the Paecilomyces and Alternaria were also present but in lower numbers. These findings indicate that fungi can tolerate very high concentrations of uranium and are more resistant than bacteria. Bacteria and fungi isolated at the Ranger LAAs from soils with high concentrations of uranium may have uranium binding capability and hence the potential for uranium bioremediation. -- Highlights: ► Fungi outcompete bacteria under increased uranium concentration in culture media. ► Soil microorganisms isolated from the Ranger Land Application Areas (LAAs) were resistant to uranium. ► Bacillus was the most abundant cultivable genus retrieved from the Ranger LAAs soils. ► Uranium in LAAs soils is

  6. 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.

  7. 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

  8. 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

  9. Contribution of Co2+ in increasing chlorophyll a concentration of Nannochloropsis salina in controlled Conwy medium

    Science.gov (United States)

    Hala, Y.; Taba, P.; Suryati, E.; Kasih, P.; Firman, N. F.

    2018-03-01

    A research in determining the contribution of Co2+ on the increase of chlorophyll a concentration of Nannochloropsis salina has been caried out. The cultivation of N. salina was conducted in the Conwy medium with a salinity of 5%o and 25%o and various Co2+ concentration (2, 4, and 8 ppm). In this research, Co2+ was exposed early in the cultivation of N. salina. The growth of N. salina was observed daily by counting the number of populations using a haemocytometer while the chlorophyll a concentration was determined by a Uv-Vis spectrophotometer. The results showed that the growth of N. salina in the control was higher than that in the medium containing Co2+. The optimum growth time was achieved on 15th days (5%) and 8th days (25%). In the cultivation medium with a salinity of 5%, Co2+ with a concentration of 2 ppm increased the chlorophyll a level while Co2+ with concentrations of 4 and 8 ppm decreased it. In the medium of cultivation with a salinity of 25%, the increase in chlorophyll a level was observed at Co2+ concentrations of 2 and 4 ppm whereas the decrease in chlorophyl a level was given at a concentration of 8 ppm. It can be concluded that at low concentrations, Co2+ increased the concentration of chlorophyll a in N. salina.

  10. 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.

  11. 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

  12. 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.

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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

    groups. Free β-hCG (MoM) ± SEM differed significantly (1.55 ± 0.19 in Vitrolife versus 1.06 ± 0.10 in Cook; P = 0.031, Student's t-test). At 20 weeks' gestation, a more advanced GA, reflecting an increased fetal growth, was seen at ultrasound examination in the Vitrolife group (n = 115) when compared with the Cook group (n = 91). After adjustment for confounding factors, both the difference between GA based on three biparietal diameter dating formulas minus the actual (ovum retrieval based) GA (adjusted mean difference + 1.14 days (P = 0.04), +1.14 days (P = 0.04) and +1.36 days (P = 0.048)), as well as head circumference (HC) and trans-cerebellar diameter (TCD) were significantly higher in the Vitrolife group (HCvl 177.3 mm, HCc 175.9 mm, adjusted mean difference 1.8, P = 0.03; TCDvl 20.5 mm, TCDc 20.2 mm, adjusted mean difference 0.4, P = 0.008). A first trimester (12 weeks) fetal screening was not yet offered routinely during the study period, therefore only 28% of women in our study participated in this elective screening programme. Although all sonographers were experienced and specially trained to perform these ultrasound examinations and were unaware of the randomization procedure, we cannot totally rule out possible intra- and inter-observer variability. Despite being indispensable in daily practice, sonographic weight formulas have a limited accuracy. According to the fetal origins hypothesis, many adult diseases originate in utero owing to adaptations made by the fetus to the environment it encounters. This study indicates that the embryonic environment is already important for fetal development. Therefore, our study emphasizes the need to investigate fetal growth patterns after assisted reproduction technologies and long-term health outcomes of IVF children, especially in relation to the culture medium used during the first few days of preimplantation development. Not applicable.

  18. A method to evaluate the dose increase in CT with iodinated contrast medium

    International Nuclear Information System (INIS)

    Amato, Ernesto; Lizio, Domenico; Settineri, Nicola; Di Pasquale, Andrea; Salamone, Ignazio; Pandolfo, Ignazio

    2010-01-01

    Purpose: The objective of this study is to develop a method to calculate the relative dose increase when a computerized tomography scan (CT) is carried out after administration of iodinated contrast medium, with respect to the same CT scan in absence of contrast medium. Methods: A Monte Carlo simulation in GEANT4 of anthropomorphic neck and abdomen phantoms exposed to a simplified model of CT scanner was set up in order to calculate the increase of dose to thyroid, liver, spleen, kidneys, and pancreas as a function of the quantity of iodine accumulated; a series of experimental measurements of Hounsfield unit (HU) increment for known concentrations of iodinated contrast medium was carried out on a Siemens Sensation 16 CT scanner in order to obtain a relationship between the increment in HU and the relative dose increase in the organs studied. The authors applied such a method to calculate the average dose increase in three patients who underwent standard CT protocols consisting of one native scan in absence of contrast, followed by a contrast-enhanced scan in venous phase. Results: The authors validated their GEANT4 Monte Carlo simulation by comparing the resulting dose increases for iodine solutions in water with the ones presented in literature and with their experimental data obtained through a Roentgen therapy unit. The relative dose increases as a function of the iodine mass fraction accumulated and as a function of the Hounsfield unit increment between the contrast-enhanced scan and the native scan are presented. The data shown for the three patients exhibit an average relative dose increase between 22% for liver and 74% for kidneys; also, spleen (34%), pancreas (28%), and thyroid (48%) show a remarkable average increase. Conclusions: The method developed allows a simple evaluation of the dose increase when iodinated contrast medium is used in CT scans, basing on the increment in Hounsfield units observed on the patients' organs. Since many clinical protocols

  19. A method to evaluate the dose increase in CT with iodinated contrast medium

    Energy Technology Data Exchange (ETDEWEB)

    Amato, Ernesto; Lizio, Domenico; Settineri, Nicola; Di Pasquale, Andrea; Salamone, Ignazio; Pandolfo, Ignazio [Department of Radiological Sciences, University of Messina, Messina 98125 (Italy); Department of Physics, University of Messina, Messina 98166 (Italy); University Hospital ' ' G. Martino' ' , Messina 98125 (Italy); Department of Radiological Sciences, University of Messina, Messina 98125 (Italy) and University Hospital ' ' G. Martino' ' , Messina 98125 (Italy)

    2010-08-15

    Purpose: The objective of this study is to develop a method to calculate the relative dose increase when a computerized tomography scan (CT) is carried out after administration of iodinated contrast medium, with respect to the same CT scan in absence of contrast medium. Methods: A Monte Carlo simulation in GEANT4 of anthropomorphic neck and abdomen phantoms exposed to a simplified model of CT scanner was set up in order to calculate the increase of dose to thyroid, liver, spleen, kidneys, and pancreas as a function of the quantity of iodine accumulated; a series of experimental measurements of Hounsfield unit (HU) increment for known concentrations of iodinated contrast medium was carried out on a Siemens Sensation 16 CT scanner in order to obtain a relationship between the increment in HU and the relative dose increase in the organs studied. The authors applied such a method to calculate the average dose increase in three patients who underwent standard CT protocols consisting of one native scan in absence of contrast, followed by a contrast-enhanced scan in venous phase. Results: The authors validated their GEANT4 Monte Carlo simulation by comparing the resulting dose increases for iodine solutions in water with the ones presented in literature and with their experimental data obtained through a Roentgen therapy unit. The relative dose increases as a function of the iodine mass fraction accumulated and as a function of the Hounsfield unit increment between the contrast-enhanced scan and the native scan are presented. The data shown for the three patients exhibit an average relative dose increase between 22% for liver and 74% for kidneys; also, spleen (34%), pancreas (28%), and thyroid (48%) show a remarkable average increase. Conclusions: The method developed allows a simple evaluation of the dose increase when iodinated contrast medium is used in CT scans, basing on the increment in Hounsfield units observed on the patients' organs. Since many clinical

  20. 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

  1. Can One Undergraduate Course Increase Cross-Cultural Competence?

    Science.gov (United States)

    Spitzer, Lois

    2015-01-01

    The majority of students who took this general education undergraduate course in developing cross-cultural understanding at a state college in the northeastern United States reported that their level of cross-cultural competence and global awareness increased by the end of the course. The primary course objective was to help students better…

  2. 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.

  3. 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 ...

  4. 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.

  5. 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

  6. 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

  7. High selection pressure promotes increase in cumulative adaptive culture.

    Directory of Open Access Journals (Sweden)

    Carolin Vegvari

    Full Text Available The evolution of cumulative adaptive culture has received widespread interest in recent years, especially the factors promoting its occurrence. Current evolutionary models suggest that an increase in population size may lead to an increase in cultural complexity via a higher rate of cultural transmission and innovation. However, relatively little attention has been paid to the role of natural selection in the evolution of cultural complexity. Here we use an agent-based simulation model to demonstrate that high selection pressure in the form of resource pressure promotes the accumulation of adaptive culture in spite of small population sizes and high innovation costs. We argue that the interaction of demography and selection is important, and that neither can be considered in isolation. We predict that an increase in cultural complexity is most likely to occur under conditions of population pressure relative to resource availability. Our model may help to explain why culture change can occur without major environmental change. We suggest that understanding the interaction between shifting selective pressures and demography is essential for explaining the evolution of cultural complexity.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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...

  18. Implementing Mobile Virtual Exhibition to Increase Cultural Heritage Visibility

    Directory of Open Access Journals (Sweden)

    Cristian CIUREA

    2014-01-01

    Full Text Available The paper presents an implementation of iOS mobile application designed as a virtual exhibition, which aims to increase the accessibility and visibility of physical objects that composite cultural heritage elements. Mobile technologies have seen a huge evolution in the last years and people are very attracted by smartphones, tablets and other mobile devices. Taking into consideration the impact of mobile technologies in all the activity fields, an important research objective is to analyze the influence of mobile applications designed as virtual exhibitions on cultural heritage promotion and on people cultural needs.

  19. 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)

  20. 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

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  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. 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

  11. Method for increasing the lifetime of an extraction medium used for reprocessing spent nuclear fuel and/or breeder materials

    International Nuclear Information System (INIS)

    Schmieder, H.; Stieglitz, L.

    1977-01-01

    A method is provided for increasing the lifetime of an extraction medium containing an organophosphorus acid ester and a hydrocarbon and being used for reprocessing spent nuclear fuel and/or breeder materials. Impurities resulting from chemical and/or radiolytic decomposition and interfering compounds of such impurities with radionuclides are removed from the extraction medium by bringing the extraction medium, after use, into intimate contact with an aqueous hydrazine hydrate solution having a concentration of between 0.1 and 1.0 molar at a temperature between 20 to 75 0 C. The aqueous hydrazine hydrate solution is then separated from the extraction medium

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Norepinephrine metabolism in neuronal cultures is increased by angiotensin II

    International Nuclear Information System (INIS)

    Sumners, C.; Shalit, S.L.; Kalberg, C.J.; Raizada, M.K.

    1987-01-01

    In this study the authors have examined the actions of angiotensin II (ANG II) on catecholamine metabolism in neuronal brain cell cultures prepared from the hypothalamus and brain stem. Neuronal cultures prepared from the brains of 1-day-old Sprague-Dawley rats exhibit specific neuronal uptake mechanisms for both norepinephrine (NE) and dopamine (DA), and also monoamine oxidase (MAO) and catechol O-methyltransferase (COMT) activity. Separate neuronal uptake sites for NE and DA were identified by using specific neuronal uptake inhibitors for each amine. In previous studies, they determined that ANG II (10 nM-1 μM) stimulates increased neuronal [ 3 H]NE uptake by acting as specific receptors. They have confirmed these results here and in addition have shown that ANG II has not significant effects on neuronal [ 3 H]DA uptake. These results suggest that the actions of ANG II are restricted to the NE transporter in neuronal cultures. It is possible that ANG II stimulates the intraneuronal metabolism of at least part of the NE that is taken up, because the peptide stimulates MAO activity, an effect mediated by specific ANG II receptors. ANG II had no effect on COMT activity in neuronal cultures. Therefore, the use of neuronal cultures of hypothalamus and brain stem they have determined that ANG II can specifically alter NE metabolism in these areas, while apparently not altering DA metabolism

  2. 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.

  3. 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

  4. 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

  5. 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.

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

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    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

  7. 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.

  8. 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.

  9. 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.

  10. 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)

  11. 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.

  12. Effect of glycine and alanine supplementation on development of cattle embryos cultured in CR1aa medium with or without cumulus cells

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    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.;

  13. 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.

  14. 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.

  15. Increasing the production of desulfurizing biocatalysts by means of fed - batch culture

    International Nuclear Information System (INIS)

    Berdugo, C I; Mena, J A; Acero, J R; Mogollon, L

    2001-01-01

    Over the past years, environmental regulations have driven a lot of effort for the development of new technologies for the upgrading of fossil fuels. Biotechnology offers an alternative way to process fossil fuels by means of a biodesulfurization technology where the production of the biocatalyst is one of the key topics. Traditionally, the production is carried out in batch culture where the maximum cellular concentration is restricted by inherent limitations of the culture type and the microorganism growth rate. This work addresses the production of two desulfurizing microorganisms: Rhodococcus erythropolis IGTS8 and gordona rubropertinctus ICP172 using fed-batch culture. Fed-batch cultures were conducted in a 12 L fermentor using ICP 4 medium containing glucose and DMSO as carbon and sulfur sources. As a result, cell concentration was increased 1.5 and 3 times with fed-batch cultures using constant and exponential flow respectively, achieving a maximum cell concentration of 7.3 g DCW/L of biocatalyst igts8 and 12.85 gGDCW/L of the new biocatalyst ICP172. Both biocatalysts presented biodesulfurization activity in a spiked matrix DBT/HXD and in diesel matrix with the detection of 2-HBP which is the end-product of DBT degradation pathway

  16. 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.

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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)....

  4. 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)

  5. 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)

  6. 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.

  7. 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.

  8. 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.

  9. 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...

  10. 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.

  11. 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...

  12. 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

  13. Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

    Science.gov (United States)

    Stampfer, Martha R; Garbe, James C

    2015-02-24

    Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

  14. 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

  15. 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.

  16. 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.

  17. 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%.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. Progranulin increases phagocytosis by retinal pigment epithelial cells in culture.

    Science.gov (United States)

    Murase, Hiromi; Tsuruma, Kazuhiro; Kuse, Yoshiki; Shimazawa, Masamitsu; Hara, Hideaki

    2017-12-01

    Retinal pigment epithelium (RPE) cells take part in retinal preservation, such as phagocytizing the shed photoreceptor outer segments (POS), every day. The incomplete phagocytic function accelerates RPE degeneration and formation of the toxic by-product lipofuscin. Excessive lipofuscin accumulation is characteristic of various blinding diseases in the human eye. Progranulin is a cysteine-rich protein that has multiple biological activities, and it has a high presence in the retina. Progranulin has been recognized to be involved in macrophage phagocytosis in the brain. The purpose of this study is to determine whether progranulin influences phagocytosis by RPE cells. All experiments were performed on primary human RPE (hRPE) cells in culture. pHrodo was used to label the isolated porcine POS, and quantification of pHrodo fluorescence was used to determine the degree of phagocytosis. Western blotting and immunohistochemistry of key proteins involved in phagocytosis were used to clarify the mechanism of progranulin. Progranulin increased RPE phagocytosis in hydrogen peroxide-treated and nontreated RPE cells. The phosphorylated form of Mer tyrosine kinase, which is important for POS internalization, was significantly increased in the progranulin-exposed cells. This increase was attenuated by SU11274, an inhibitor of hepatic growth factor receptor. Under the oxidative stress condition, exposure to progranulin led to an approximately twofold increase in integrin alpha-v, which is associated with the first step in recognition of POS by RPE cells. These results suggest that progranulin could be an effective stimulator for RPE phagocytosis and could repair RPE function. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  4. 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...

  5. 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.

  6. 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.

  7. 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.

  8. 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...

  9. 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.

  10. Using Six-Word Memoirs to Increase Cultural Identity Awareness

    Science.gov (United States)

    Simmons, Nathaniel; Chen, Yea-Wen

    2014-01-01

    Guided by cultural identity theory (CIT), the authors offer the six-word memoir (6WM) as a storytelling vehicle to engage students in critical, reflexive (re)considerations of their cultural identities and positions. This activity's impetus is threefold. First, it recognizes the practical challenges of teaching and learning the important, yet…

  11. Editorial : initiating cultural heritage research to increase Europe's competitiveness

    NARCIS (Netherlands)

    Pereira Roders, A.R.; Oers, van R.

    2011-01-01

    Purpose – The purpose of this paper is to present and discuss the contribution of European Commission (EC) initiatives to stimulate cultural heritage research over the last 20 years and the contribution of the research results to cultural heritage management and sustainable development.

  12. Follicle stimulating hormone increases spermatogonial stem cell colonization during in vitro co-culture.

    Science.gov (United States)

    Narenji Sani, Reza; Tajik, Parviz; Yousefi, Mohammad Hassan; Movahedin, Mansoureh; Qasemi-Panahi, Babak; Shafiei, Shiva; Ahmadi Hamedani, Mahmood

    2013-01-01

    The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells (SCCs) have provided very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist of spermatogonial stem cells. To investigate and biomanipulation of these cells, proliferation and viability rate of cells should be increased in vitro, at first. Follicle stimulating hormone (FSH) has been suggested to play a determinant role in the survival of germ cells in addition to increasing spermatogonial proliferation. In this study, the in vitro effects of FSH on spermatogonial cell colony formation were investigated. Sertoli and spermatogonial cells were isolated from 3-5 months old calves. The identity of the Sertoli cells and spermatogonial stem cells were confirmed through immunocytochemistry and colony morphology, respectively. Co-cultured Sertoli and spermatogonial cells were treated with FSH in different dose of 10, 20 and 40 IU mL(-1) FSH, before colony assay. Results indicated that, FSH increased in vitro colonization of spermatogonial cells in comparison with control group. In conclusion, using FSH provided proper bovine spermatogonial stem cell culture medium for in vitro study of these cells.

  13. 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

  14. 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.

  15. 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

  16. Exogenous glutamine increases lipid accumulation in developing seeds of castor bean (Ricinus communis L. cultured in vitro

    Directory of Open Access Journals (Sweden)

    Zhang Yang

    2015-01-01

    Full Text Available This report describes biomass production and compositional changes of developing castor seeds in response to change in the nitrogen resource (glutamine of the medium. During the early developmental period (24-36 days after pollination, oil was found to initially accumulate in the developing seeds. Carbohydrates and oil were inversely related after glutamine provision (35 mM, in the culture medium. [U-14C] sucrose labeling was used to investigate the effect of metabolic fluxes among different storage materials. Addition of glutamine led to a 7% increase of labeling in lipids and an inverse decrease of labeling in carbohydrates. It was postulated that changes in the glutamine concentration in the medium are likely to influence the partitioning of resources between the various storage products, especially carbohydrates and oil. These observations will contribute to a better understanding of assimilate partitioning in developing castor seeds and the development of molecular strategies to improve castor bean seed quality and plant breeding studies.

  17. 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.

  18. 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.

  19. 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

  20. Medium-chain, triglyceride-containing lipid emulsions increase human neutrophil beta2 integrin expression, adhesion, and degranulation.

    Science.gov (United States)

    Wanten, G J; Geijtenbeek, T B; Raymakers, R A; van Kooyk, Y; Roos, D; Jansen, J B; Naber, A H

    2000-01-01

    To test the hypothesis that lipid emulsions with different triglyceride structures have distinct immunomodulatory properties, we analyzed human neutrophil adhesion and degranulation after lipid incubation. Neutrophils, isolated from the blood of 10 healthy volunteers, were incubated in medium or physiologic (2.5 mmol/L) emulsions containing long-chain (LCT), medium-chain (MCT), mixed LCT/MCT, or structured (SL) triglycerides. Expression of adhesion molecules and degranulation markers was evaluated by flow cytometry. Also, functional adhesion was investigated by means of a flow cytometric assay using fluorescent beads coated with the integrin ligand intercellular adhesion molecule (ICAM)-1. Although LCT and SL had no effect, LCT/MCT significantly increased expression of the beta2 integrins lymphocyte-function-associated antigen 1 (+18%), macrophage antigen 1 (+387%), p150,95 (+82%), and (alphaDbeta2 (+230%). Degranulation marker expression for azurophilic (CD63, +210%) and specific granules (CD66b, +370%) also significantly increased, whereas L-selectin (CD62L, -70%) decreased. The effects of LCT/MCT were mimicked by the MCT emulsion. ICAM-1 adhesion (% beads bound) was increased by LCT/MCT (34% +/- 4%), whereas LCT (19% +/-3%) and SL (20% +/- 2%) had no effect compared with medium (17% +/- 3%). LCT/MCT and MCT, contrary to LCT and SL emulsions, increased neutrophil beta2 integrin expression, adhesion, and degranulation. Apart from other emulsion constituents, triglyceride chain length might therefore be a key feature in the interaction of lipid emulsions and the phagocyte immune system.

  1. 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

  2. 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.

  3. 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...

  4. 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.

  5. 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

  6. 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.

  7. 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).

  8. 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…

  9. 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

  10. Study Abroad in Psychology: Increasing Cultural Competencies through Experiential Learning

    Science.gov (United States)

    Earnest, David R.; Rosenbusch, Katherine; Wallace-Williams, Devin; Keim, Alaina C.

    2016-01-01

    Despite the prominence of study abroad programs, few are offered in the field of psychology. The current study sought to investigate the impact of study abroad programs in psychology through a comparison of study abroad and domestic student cultural competencies. Participants included 104 undergraduate students enrolled in either a psychology…

  11. Mixed cultures of Kimchi lactic acid bacteria show increased cell ...

    African Journals Online (AJOL)

    ufuoma

    anaerobic organisms that are highly resistant to salts. Probiotic cultures for use in ... kimchi have a superior ability to decompose and utilize nutrients, and show ... citrate, 5 g sodium acetate, 1 g Tween, 2 g K2HPO4, 0.2 g. MgSO4•7H2O, 0.2 g ...

  12. Did Cultural and Artistic Education in the Netherlands increase Student Participation in High Cultural Events?

    NARCIS (Netherlands)

    Damen, Marie Louise; Van Klaveren, Chris

    2013-01-01

    This study examines whether Cultural and Artistic Education in the Netherlands caused students to participate more in high cultural events. A unique feature of the intervention was that students were free to choose the type of cultural event they participated in. So the intervention relied on the

  13. Did cultural and artistic education in the Netherlands increase student participation in high cultural events?

    NARCIS (Netherlands)

    Damen, M.-L.; van Klaveren, C.

    2010-01-01

    This study examines whether Cultural and Artistic Education that was implemented by the Dutch Ministry of Education, Culture and Science in 1999 caused students to participate more in high cultural events. A unique feature of the intervention was that students were free to choose the type of

  14. Follicle stimulating hormone increases spermatogonial stem cell colonization during in vitro co - culture

    Directory of Open Access Journals (Sweden)

    Reza Narenji Sani

    2013-03-01

    Full Text Available The complex process of spermatogenesis is regulated by various factors. Studies onspermatogonial stem cells(SCCshave provided very important tool to improve herd geneticand different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist ofspermatogonial stem cells. To investigate and biomanipulation of these cells, proliferationand viability rate of cells should be increasedin vitro, at first. Follicle stimulating hormone(FSH has been suggested to play a determinant role in the survival of germ cells in additionto increasing spermatogonial proliferation. In this study, thein vitroeffects ofFSHonspermatogonial cell colony formation were investigated. Sertoli and spermatogonial cellswere isolated from 3-5 months old calves. The identity of theSertoli cells and spermatogonialstem cells were confirmed through immunocytochemistry and colony morphology,respectively. Co-cultured Sertoli and spermatogonial cells were treatedwithFSHin differentdose of10, 20 and 40 IU mL-1FSH, before colony assay.Results indicated that,FSHincreasedin vitrocolonization of spermatogonial cells in comparison with control group. In conclusion,usingFSHprovided proper bovine spermatogonial stem cell culture medium forin vitrostudy of these cells.

  15. 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.

  16. 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

  17. 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.

  18. How Universities Can Increase Enrollment by Advertising Internships: The "Message" and the "Medium"

    Science.gov (United States)

    Tucciarone, Kristy

    2015-01-01

    This study investigates how universities can increase enrollment by advertising internships to prospective students during the college search process. The primary reason students earn a college degree is to secure a good-quality career with earning potential. Internships--the single most important credential for recent graduates--are the key…

  19. 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

  20. 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

  1. Meeting increased logistical demands : Developing as a small- and medium-sized system supplier

    OpenAIRE

    Carlsson, Inga-Lill

    2009-01-01

    Many subcontractors choose to implement a strategy of “system supply” in order to meetincreasing global competition. They are then confronted with increased demands to take agreater overall responsibility in this role. It is important to investigate the implications of theseresponsibilities before investing in developing the organization, especially for a small- ormedium-sized subcontractor with limited resources. The customer’s view of different demandsdoes not necessarily correspond to how ...

  2. 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.

  3. 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.

  4. THE STRATEGY OF EMPOWERMENT SMALL AND MEDIUM ENTERPRISES IN THE EFFORT FOR INCREASING REVENUE AND MARKET

    Directory of Open Access Journals (Sweden)

    PRASETIO ARIWIBOWO

    2018-04-01

    Full Text Available The development of SMEs sector, explicitly aimed at the efforts to realize the power of the nation-competitiveness in order to strengthen the global competitive economy. This research uses qualitative descriptive method to describe each variable analyzed. The analysis is SWOT analysis and Analytical Hierarchy Procees (AHP to calculate any case the most preferred and influence the development of the empowerment of SMEs especially in terms of revenue and market expansion in Semarang based and questionnaire that sample scrambled by the author. Objectives and results of this study is to determine the priority of the strategy used by the Semarang city government and become the main reference for entrepreneurs of SMEs in Semarang to increase revenue and market share in the city of Semarang is improving managerial skills by improving capacity, work ethic, and the performance of the institution and the staff of the local government of Semarang, and increase community participation in the planning, implementation, and monitoring of development of SMEs.

  5. 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.

  6. 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.

  7. Response of Xylella fastidiosa to zinc: decreased culturability, increased exopolysaccharide production, and formation of resilient biofilms under flow conditions.

    Science.gov (United States)

    Navarrete, Fernando; De La Fuente, Leonardo

    2014-02-01

    The bacterial plant pathogen Xylella fastidiosa produces biofilm that accumulates in the host xylem vessels, affecting disease development in various crops and bacterial acquisition by insect vectors. Biofilms are sensitive to the chemical composition of the environment, and mineral elements being transported in the xylem are of special interest for this pathosystem. Here, X. fastidiosa liquid cultures were supplemented with zinc and compared with nonamended cultures to determine the effects of Zn on growth, biofilm, and exopolysaccharide (EPS) production under batch and flow culture conditions. The results show that Zn reduces growth and biofilm production under both conditions. However, in microfluidic chambers under liquid flow and with constant bacterial supplementation (closer to conditions inside the host), a dramatic increase in biofilm aggregates was seen in the Zn-amended medium. Biofilms formed under these conditions were strongly attached to surfaces and were not removed by medium flow. This phenomenon was correlated with increased EPS production in stationary-phase cells grown under high Zn concentrations. Zn did not cause greater adhesion to surfaces by individual cells. Additionally, viability analyses suggest that X. fastidiosa may be able to enter the viable but nonculturable state in vitro, and Zn can hasten the onset of this state. Together, these findings suggest that Zn can act as a stress factor with pleiotropic effects on X. fastidiosa and indicate that, although Zn could be used as a bactericide treatment, it could trigger the undesired effect of stronger biofilm formation upon reinoculation events.

  8. 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.

  9. 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)

  10. 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.

  11. 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.

  12. Increased production of outer membrane vesicles by cultured freshwater bacteria in response to ultraviolet radiation.

    Science.gov (United States)

    Gamalier, Juliana P; Silva, Thiago P; Zarantonello, Victor; Dias, Felipe F; Melo, Rossana C N

    2017-01-01

    Secretion of membrane vesicles is an important biological process of both eukaryotic and prokaryotic cells. This process has been characterized in pathogenic bacteria, but is less clear in non-pathogenic bacteria from aquatic ecosystems. Here, we investigated, for the first time, the process of formation of outer membranes vesicles (OMVs), nanoscale vesicles extruded from the outer membrane (OM) of gram-negative bacteria, in cultures of freshwater bacteria after exposure or not to ultraviolet radiation (UVR) as an environmental stressor. Non-axenic cultures of freshwater bacteria isolated from a Brazilian aquatic ecosystem (Funil reservoir) were exposed or not to UVR (UVA+UVB) over a 3h period, during which cell density, viability and ultrastructure were analyzed. First, we showed that UVR induce bacterial death. UVR triggered significant negative effect on cell density after 3h of UVR treatment. This decrease was directly associated with cell death as revealed by a cell viability fluorescent probe that enables the distinction of live/dead bacteria. Transmission electron microscopy (TEM) revealed changes indicative of cell death after 3h of UVR exposure, with significant increase of damaged cells compared to the control group. Second, we demonstrated that gram-negative bacteria release OMVs during normal growth and after UVR exposure. OMVs were clearly identified as round, membrane-bound vesicles budding off from the bacterial OM as isolated or clustered vesicles or free in the extracellular medium. Remarkably, quantitative TEM analyses showed that bacteria respond to UVR with increased formation of OMVs. Moreover, while OMVs numbers per intact or damaged cell did not differ in the untreated group, UVR led to a higher vesiculation by bacteria in process of death. This means that degenerating bacteria release OMVs before lysis and that this secretion might be an adaptive/protective response to rapid changes in environmental conditions such as UV radiation. Copyright

  13. 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.

  14. Mechanical stimulation increases proliferation, differentiation and protein expression in culture

    DEFF Research Database (Denmark)

    Grossi, Alberto; Yadav, Kavita; Lawson, Moira Ann

    2007-01-01

    Myogenesis is a complex sequence of events, including the irreversible transition from the proliferation-competent myoblast stage into fused, multinucleated myotubes. Myogenic differentiation is regulated by positive and negative signals from surrounding tissues. Stimulation due to stretch- or load...... to elucidate also the signaling pathway by which this mechanical stimulation can causes an increase in protein expression. When mechanically stimulated via laminin receptors on cell surface, C(2)C(12) cells showed an increase in cell proliferation and differentiation. Populations undergoing mechanical...... stimulation through laminin receptors show an increase in expression of Myo-D, myogenin and an increase in ERK1/2 phosphorylation. Cells stimulated via fibronectin receptors show no significant increases in fusion competence. We conclude that load induced signalling through integrin containing laminin...

  15. 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.

  16. Non-face-to face international business negotiation : how is national culture reflected in this medium?

    NARCIS (Netherlands)

    Ulijn, J.M.; Lincke, A.; Karakaya, Y.

    2001-01-01

    With the globalization of the world economy, it is imperative that managers, both present and future, be sensitive to differences in intercultural business communication. In particular, the context of global electronic commerce leads to an increasing use of email in negotiating deals, which to this

  17. Electronic business negotiation : some experimental studies on the interaction between medium, innovation context and culture

    NARCIS (Netherlands)

    Lincke, A.M.Th.

    2003-01-01

    Negotiating a deal with a new client, customer, or supplier has traditionally meant meeting in person, sometimes enduring long hours in transit. Today, business people find themselves with an ever-increasing array of technologies for communicating and initiating relationships, many of which do not

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Assessment of Chlorella vulgaris and indigenous microalgae biomass with treated wastewater as growth culture medium.

    Science.gov (United States)

    Fernández-Linares, Luis C; Guerrero Barajas, Claudia; Durán Páramo, Enrique; Badillo Corona, Jesús A

    2017-11-01

    The aim of the present work was to evaluate the feasibility of microalgae cultivation using secondary treated domestic wastewater. Two Chlorella vulgaris strains (CICESE and UTEX) and an indigenous consortium, were cultivated on treated wastewater enriched with and without the fertilizer Bayfolan®. Biomass production for C. vulgaris UTEX, CICESE and the indigenous consortium grown in treated wastewater was 1.167±0.057, 1.575±0.434 and 1.125±0.250g/L, with a total lipid content of 25.70±1.24, 23.35±3.01and 20.54±1.23% dw, respectively. The fatty acids profiles were mainly composed of C16 and C18. Regardless of the media used, in all three strains unsaturated fatty acids were the main FAME (fatty acids methyl esters) accumulated in a range of 45-62%. An enrichment of treated wastewater with Bayfolan® significantly increased the production of biomass along with an increase in pigments and proteins of ten and threefold, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. A study of the irradiation of sawdust culture medium and the yield of fresh lentinus

    International Nuclear Information System (INIS)

    Xu Meiyu; Wu Jinshui

    1989-01-01

    60 Co γ-rays at the doses of 1.0-1.8 Mrad were used to irradiate sawdust material for sterilization, It was found that 60 Co γ-rays could promote the breakdown of cellulose into soluble sugar, and thus promoted the growth of hyphae and increased the yield of L.edode. Determination of regressive analysis and correlation coefficients indicated that there was a high positive correlation between soluble sugar and the growth of hyphae(r = 0.9546**) , which can be expressed as Y = 3.12 + 1.30X; A high positive correlation between soluble sugar and the yield of L.edode(r = 0.9935**) can be expressed as Y = 31.95 + 86.69X; and a high positive correlation between the hyphae growing rate and the yield of L.edode(r = 0.9531**) can be expressed as Y = -145.34 + 60.92X

  4. [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.

  5. 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)

  6. Does a Culture of Happiness Increase Rumination Over Failure?

    Science.gov (United States)

    McGuirk, Lucy; Kuppens, Peter; Kingston, Rosemary; Bastian, Brock

    2017-07-17

    Promoting happiness within society is good for health, but could the overpromotion of happiness have a downside? Across 2 studies, we investigate 2 emotion norms associated with an emphasis on happiness-the importance of (a) seeking positive emotion, and (b) avoiding negative emotion-and whether these norms have implications for how people respond to, and seek to regulate, their negative emotional experiences. In Study 1, we used an experimental design to show that emphasizing the importance of happiness increased rumination in response to failure. In Study 2, we drew on cross-sectional evidence to investigate the other side of this equation, finding that emphasizing the importance of not experiencing negative emotional states (e.g., depression and anxiety) was also associated with increased rumination, and that this had downstream consequences for well-being. Together, the findings suggest that the overpromotion of happiness, and, in turn, the felt social pressure not to experience negative emotional states, has implications for maladaptive responses to negative emotional experiences. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  7. Medium-chain triglycerides and conjugated linoleic acids in beverage form increase satiety and reduce food intake in humans.

    Science.gov (United States)

    Coleman, Hannah; Quinn, Paul; Clegg, Miriam E

    2016-06-01

    Both developed and developing countries are seeing increasing trends of obesity in people young and old. It is thought that satiety may play a role in the prevention of obesity by increasing satiety and reducing energy intake. We hypothesized that medium-chain triglycerides (MCT) would increase satiety and decrease food intake compared with conjugated linoleic acid (CLA) and a control oil. Nineteen healthy participants were tested on 3 separate occasions, where they consumed a beverage test breakfast containing (1) vegetable oil (control), (2) CLA, or (3) MCT. Participants self-requested an ad libitum sandwich buffet lunch. Time between meals, satiety from visual analog scales, energy intake at lunch, and intake for the rest of the day using weighed food diaries were measured. The results indicated that the time until a meal request was significantly different between the 3 meals (P=.016); however, there were no differences in intakes at the ad libitum lunch (P>.05). The CLA breakfast generated the greatest delay in meal time request. There was a difference between the control lipid compared with both the CLA and MCT for energy intake over the remainder of the test day and for total energy intake on the test day (P.05). Both CLA and MCT increased satiety and reduced energy intake, indicating a potential role in aiding the maintenance of energy balance. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. 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.

  9. 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)

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. The relationship between cultural competence education and increasing diversity in nursing schools and practice settings.

    Science.gov (United States)

    Pacquiao, Dula

    2007-01-01

    This article attempted to examine the relationship between cultural competence education and increasing diversity in nursing schools and practice settings. In addition to the review of the literature, a panel of experts was interviewed regarding institutional practices in response to the challenge of increasing diversity and cultural competence education. Evidence of positive outcomes of cultural competent care and impact of race and ethnic concordance between patients and providers are presented. The challenge of increasing underrepresented minorities in health care professions remains elusive. An ecological analysis is recommended to address the social and cultural barriers that transcend the micro system of the school and the macro system of the society. The challenge of increasing diversity and realizing outcomes of cultural competence education requires social and comprehensive remedies to level life inequities that perpetuate a history of disadvantages in some groups.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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).

  20. 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.

  1. A Systematic Review: The Next Generation Science Standards and the Increased Cultural Diversity

    Science.gov (United States)

    Asowayan, Alaa A.; Ashreef, Samaar Y.; Omar, Sozan H.

    2017-01-01

    This systematic review aims to explore the effect of NGSS on students' academic excellence. Specifically, considering increased cultural diversity, it is appropriate to identify student's science-related values, respectful features of teachers' cultural competence, and underlying challenges and detect in what ways these objectives are addressed by…

  2. Fish culture in heated effluents (eastern Europe). [Growth increase in carp and rainbow trout

    Energy Technology Data Exchange (ETDEWEB)

    Backiel, T; Gay, B; La Croix, M J.B.; Ophel, I L [eds.

    1976-01-01

    Dependence of growth and reproduction of cultivated fishes on temperature are briefly reviewed. Experiments with and commercial-size operations of various methods of fish culture with the use of heated effluents are exemplified. Cage culture, earthen ponds, flowing water culture have demonstrated possibilities of growth increase in carp and rainbow trout and of controlled reproduction of the former. Problems involved in utilization of various systems and several points of view on the use of low-grade heat for fish culture are considered.

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. 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...

  8. 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.

  9. 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.

  10. Increased Arginine and Ornithine Flux in Islets of Langerhans Cultured in a Microgravity Model System

    Science.gov (United States)

    Tobin, B. W.; Sams, C. F.; Smith, S. M.

    2000-01-01

    Microgravity is associated with alterations in protein metabolism of both muscle and bone. That pancreas-derived insulin is essential to the normal maintenance of body protein balance is well known. The importance of altered endocrine pancreas function in microgravity is not yet established. We proposed to examine the influence of a microgravity model system, the High Aspect Ratio Vessel (HARV) upon islets of Langerhans from Wistar Furth rats. Islets were cultured in the HARV for 48 hr in Medium-199 and contrasted to static control islets (PLATE). Nitrogenous compounds elaborated into the media (micromoles/ml) were analyzed at 0 and 48 hr of culture and compared to PLATE with a 2-way ANOVA (HARV vs Hour).

  11. 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.

  12. Structured medium and long chain triglycerides show short-term increases in fat oxidation, but no changes in adiposity in men.

    Science.gov (United States)

    Roynette, Catherine E; Rudkowska, Iwona; Nakhasi, Dilip K; Jones, Peter J H

    2008-05-01

    Medium chain triglycerides (MCT) have been suggested as modulators of human energy expenditure (EE) and thus may influence total and regional body fat distribution. To investigate in overweight men the effects of structured medium and long chain triglycerides on EE, substrate oxidation and body adiposity, compared to extra virgin olive oil (OO). In a 6 week single-blind crossover study, 23 overweight men were randomly assigned to consume a standard high-fat diet of which 75% total fat was provided as either structured medium and long chain triglycerides referred to as structured oil (StO), or OO. EE and body composition were measured using indirect calorimetry and magnetic resonance imaging, respectively, at weeks 1 and 6 of each phase. Body weight decreased (pstructured medium and long chain triglyceride oil increases short-term fat oxidation but fails to modulate body weight or adiposity through a change in EE.

  13. Sociocultural influences on strategies to lose weight, gain weight, and increase muscles among ten cultural groups.

    Science.gov (United States)

    McCabe, Marita P; Busija, Lucy; Fuller-Tyszkiewicz, Matthew; Ricciardelli, Lina; Mellor, David; Mussap, Alexander

    2015-01-01

    This study determined how sociocultural messages to change one's body are perceived by adolescents from different cultural groups. In total, 4904 adolescents, including Australian, Chilean, Chinese, Indo-Fijian, Indigenous Fijian, Greek, Malaysian, Chinese Malaysian, Tongans in New Zealand, and Tongans in Tonga, were surveyed about messages from family, peers, and the media to lose weight, gain weight, and increase muscles. Groups were best differentiated by family pressure to gain weight. Girls were more likely to receive the messages from multiple sociocultural sources whereas boys were more likely to receive the messages from the family. Some participants in a cultural group indicated higher, and others lower, levels of these sociocultural messages. These findings highlight the differences in sociocultural messages across cultural groups, but also that adolescents receive contrasting messages within a cultural group. These results demonstrate the difficulty in representing a particular message as being characteristic of each cultural group. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. A METHOD OF INCREASING THE NUMBER OF TOURISTS IN ROMANIA: SIBIU – EUROPEAN CAPITAL OF CULTURE

    Directory of Open Access Journals (Sweden)

    Gabriela Cecilia STĂNCIULESCU

    2016-06-01

    Full Text Available The title of European Capital of Culture could help immensely a city and its surrounding region from cultural, social and economic pointd of view. Cultural activity in the city is gaining momentum and new categories of tourists are attracted here. To determine if being a cultural capital influenced or not the growth and/or decrease trend as regards the number of tourists that came to Sibiu in a consecutive period of 23 years, the starting point was that this event had a major impact on tourists. The present paper is a quantitative research based on the number of Romanian and foreign tourists accommodated in structures of tourist receptions with functions of tourist accommodation. The conclusions of this article respond to these questions: Being a European Capital of Culture attracts or not more tourists than usual? The increase in the number of tourists was proportional with the effort of the authorities involved in the project?

  15. Enrichment methodology to increase the positivity of cultures from body fluids

    Directory of Open Access Journals (Sweden)

    Alessandra Valle Daur

    Full Text Available Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially-severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1 the conventional culture method (agar plating and (2 the enrichment culture technique, using the Bact/Alert® blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents.

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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

  1. A strategy to reduce cross-cultural miscommunication and increase the likelihood of improving health outcomes.

    Science.gov (United States)

    Kagawa-Singer, Marjorie; Kassim-Lakha, Shaheen

    2003-06-01

    Encounters between physicians and patients from different cultural backgrounds are becoming commonplace. Physicians strive to improve health outcomes and increase quality of life for every patient, yet these discordant encounters appear to be a significant factor, beyond socioeconomic barriers, in creating the unequal and avoidable excess burden of disease borne by members of ethnic minority populations in the United States. Most clinicians lack the information to understand how culture influences the clinical encounter and the skills to effectively bridge potential differences. New strategies are required to expand medical training to adequately address culturally discordant encounters among the physicians, their patients, and the families, for all three may have different concepts regarding the nature of the disease, expectations about treatment, and modes of appropriate communication beyond language. The authors provide an anthropological perspective of the fundamental relationship between culture and health, and outline systemic changes needed within the social and legal structures of the health care system to reduce the risk of cross-cultural miscommunication and increase the likelihood of improving health outcomes for all populations within the multicultural U.S. society. The authors define the strengths inherent within every culture, provide a guideline for the clinician to evaluate disease and illness within its cultural context, and outline the clinical skills required to negotiate among potential differences to reach mutually desired goals for care. Last, they indicate the structural changes required in the health care setting to enable and support such practice.

  2. INCREASING CULTURALLY COMPETENT NEUROPSYCHOLOGICAL SERVICES FOR ETHNIC MINORITY POPULATIONS: A CALL TO ACTION

    Science.gov (United States)

    Mindt, Monica Rivera; Byrd, Desiree; Saez, Pedro; Manly, Jennifer

    2010-01-01

    US demographic and sociopolitical shifts have resulted in a rapidly growing need for culturally competent neuropsychological services. However, clinical neuropsychology as a field has not kept pace with the needs of ethnic minority clients. In this discussion we review: historical precedents and the limits of universalism in neuropsychology; ethical/professional guidelines pertinent to neuropsychological practice with ethnic minority clients; critical cultural considerations in neuropsychology; current disparities germane to practice; and challenges to the provision of services to racial/ethnic minority clients. We provide a call to action for neuropsychologists and related organizations to advance multiculturalism and diversity within the field by increasing multicultural awareness and knowledge, multicultural education and training, multicultural neuropsychological research, and the provision of culturally competent neuropsychological services to racial/ethnic minority clients. Lastly, we discuss strategies for increasing the provision of culturally competent neuropsychological services, and offer several resources to meet these goals. PMID:20373222

  3. 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.

  4. 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)

  5. Plant-fed versus chemicals-fed rhizobacteria of Lucerne: Plant-only teabags culture media not only increase culturability of rhizobacteria but also recover a previously uncultured Lysobacter sp., Novosphingobium sp. and Pedobacter sp.

    Science.gov (United States)

    Hegazi, Nabil A; Sarhan, Mohamed S; Fayez, Mohamed; Patz, Sascha; Murphy, Brian R; Ruppel, Silke

    2017-01-01

    In an effort to axenically culture the previously uncultivable populations of the rhizobacteria of Lucerne (Medicago sativa L.), we propose plant-only teabags culture media to mimic the nutritional matrix available in the rhizosphere. Here, we show that culture media prepared from Lucerne powder teabags substantially increased the cultivability of Lucerne rhizobacteria compared with a standard nutrient agar, where we found that the cultivable populations significantly increased by up to 60% of the total bacterial numbers as estimated by Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Cluster analysis of 16S rDNA Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) of cultivable Colony-Forming Units (CFUs) revealed a more distinct composition and separation of bacterial populations recovered on the plant-only teabags culture media than those developed on a standard nutrient agar. Further, the new plant medium gave preference to the micro-symbiont Sinorhizobium meliloti, and succeeded in isolating a number of not-yet-cultured bacteria, most closely matched to Novosphingobium sp., Lysobacter sp. and Pedobacter sp. The present study may encourage other researchers to consider moving from the well-established standard culture media to the challenging new plant-only culture media. Such a move may reveal previously hidden members of rhizobacteria, and help to further explore their potential environmental impacts.

  6. Chronic lithium treatment increased intracellular S100ß levels in rat primary neuronal culture.

    Directory of Open Access Journals (Sweden)

    Masoumeh Emamghoreishi

    2015-02-01

    Full Text Available S100ß a neurotrophic factor mainly released by astrocytes, has been implicated in the pathogenesis of bipolar disorder. Thus, lithium may exert its neuroprotective effects to some extent through S100ß. Furthermore, the possible effects of lithium on astrocytes as well as on interactions between neurons and astrocytes as a part of its mechanisms of actions are unknown. This study was undertaken to determine the effect of lithium on S100β in neurons, astrocytes and a mixture of neurons and astrocytes. Rat primary astrocyte, neuronal and mixed neuro-astroglia cultures were prepared from cortices of 18-day's embryos. Cell cultures were exposed to lithium (1mM or vehicle for 1day (acute or 7 days (chronic. RT-PCR and ELISA determined S100β mRNA and intra- and extracellular protein levels. Chronic lithium treatment significantly increased intracellular S100β in neuronal and neuro-astroglia cultures in comparison to control cultures (P<0.05. Acute and chronic lithium treatments exerted no significant effects on intracellular S100β protein levels in astrocytes, and extracellular S100β protein levels in three studied cultures as compared to control cultures. Acute and chronic lithium treatments did not significantly alter S100β mRNA levels in three studied cultures, compared to control cultures. Chronic lithium treatment increased intracellular S100ß protein levels in a cell-type specific manner which may favor its neuroprotective action. The findings of this study suggest that lithium may exert its neuroprotective action, at least partly, by increasing neuronal S100ß level, with no effect on astrocytes or interaction between neurons and astrocytes.

  7. 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.

  8. Shaping the Organizational Culture in Conditions of Increasing the Competitiveness of Enterprises

    Directory of Open Access Journals (Sweden)

    Joanna Rębisz

    2010-10-01

    Full Text Available The organizational culture is one of key factors which can influence the organizational success in building the long-lasting domination of an enterprise. The article is an attempt to introduce and at the same time to present the understanding of the culture in terms of expected bearings which can explain why organizational individuals (or entire organizations promote only the behaviors which are in accordance with the value and the mission of the enterprises. The author draws attention to the level of expectations and behavior, which is usually the result of team work. A lot of attention has also been paid to the phenomenon of crossing of two relations: organizational culture with the function of leadership. The author has also discussed the role of a manager as a means of shaping and supporting an organization culture in conditions of increasing competition.

  9. 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.

  10. 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).

  11. 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.

  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. 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

  14. 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.

  15. 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

  16. 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

  17. Tuning Chocolate Flavor through Development of Thermotolerant Saccharomyces cerevisiae Starter Cultures with Increased Acetate Ester Production

    Science.gov (United States)

    Meersman, Esther; Steensels, Jan; Struyf, Nore; Paulus, Tinneke; Saels, Veerle; Mathawan, Melissa; Allegaert, Leen; Vrancken, Gino

    2015-01-01

    Microbial starter cultures have extensively been used to enhance the consistency and efficiency of industrial fermentations. Despite the advantages of such controlled fermentations, the fermentation involved in the production of chocolate is still a spontaneous process that relies on the natural microbiota at cocoa farms. However, recent studies indicate that certain thermotolerant Saccharomyces cerevisiae cultures can be used as starter cultures for cocoa pulp fermentation. In this study, we investigate the potential of specifically developed starter cultures to modulate chocolate aroma. Specifically, we developed several new S. cerevisiae hybrids that combine thermotolerance and efficient cocoa pulp fermentation with a high production of volatile flavor-active esters. In addition, we investigated the potential of two strains of two non-Saccharomyces species that produce very large amounts of fruity esters (Pichia kluyveri and Cyberlindnera fabianii) to modulate chocolate aroma. Gas chromatography-mass spectrometry (GC-MS) analysis of the cocoa liquor revealed an increased concentration of various flavor-active esters and a decrease in spoilage-related off-flavors in batches inoculated with S. cerevisiae starter cultures and, to a lesser extent, in batches inoculated with P. kluyveri and Cyb. fabianii. Additionally, GC-MS analysis of chocolate samples revealed that while most short-chain esters evaporated during conching, longer and more-fat-soluble ethyl and acetate esters, such as ethyl octanoate, phenylethyl acetate, ethyl phenylacetate, ethyl decanoate, and ethyl dodecanoate, remained almost unaffected. Sensory analysis by an expert panel confirmed significant differences in the aromas of chocolates produced with different starter cultures. Together, these results show that the selection of different yeast cultures opens novel avenues for modulating chocolate flavor. PMID:26590272

  18. Increased Paracrine Immunomodulatory Potential of Mesenchymal Stromal Cells in Three-Dimensional Culture

    DEFF Research Database (Denmark)

    Follin, Bjarke; Juhl, Morten; Cohen, Smadar

    2016-01-01

    Mesenchymal stromal/stem cells (MSCs) have been investigated extensively through the past years, proving to have great clinical therapeutic potential. In vitro cultivation of MSCs in three-dimensional (3D) culture systems, such as scaffolds, hydrogels, or spheroids, have recently gained attention...... for tissue engineering applications. Studies on MSC spheroids demonstrated that such cultivation increased the paracrine immunomodulatory potential of the MSCs, accompanied by phenotypic alterations. In this review, we gather results from recent experimental studies on the immunomodulatory abilities of MSCs...... when cultured as spheroids or in biomaterials like scaffolds or hydrogels compared to regular two-dimensional (2D) culture and show that alterations occurring to MSCs in spheroids also occur in MSCs in biomaterials. We provide a brief description of known mechanisms of MSC immunomodulatory capacity...

  19. 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...

  20. 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.

  1. Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose.

    Science.gov (United States)

    Beltramo, E; Berrone, E; Buttiglieri, S; Porta, M

    2004-01-01

    High glucose induces pathological alterations in small and large vessels, possibly through increased formation of AGE, activation of aldose reductase and protein kinase C, and increased flux through the hexosamine pathway. We showed previously that thiamine and benfotiamine correct delayed replication and increase lactate production in endothelial cells subjected to high glucose. We now aim at verifying the effects of thiamine and benfotiamine on cell cycle, apoptosis, and expression of adhesion molecules in endothelial cells and pericytes, under high ambient glucose. Human umbilical vein endothelial cells and bovine retinal pericytes were cultured in normal (5.6 mmol/L) or high (28 mmol/L) glucose, with or without thiamine or benfotiamine, 50 or 100 micro mol/L. Apoptosis was determined by two separate ELISA methods, measuring DNA fragmentation and caspase-3 activity, respectively. Cell cycle and integrin subunits alpha3, alpha5, and beta1 concentration were measured by flow cytometry. Apoptosis was increased in high glucose after 3 days of culture, both in endothelium and pericytes. Thiamine and benfotiamine reversed such effects. Neither cell cycle traversal nor integrin concentrations were modified in these experimental conditions. Thiamine and benfotiamine correct increased apoptosis due to high glucose in cultured vascular cells. Further elucidations of the mechanisms through which they work could help set the basis for clinical use of this vitamin in the prevention and/or treatment of diabetic microangiopathy. Copyright 2004 John Wiley & Sons, Ltd.

  2. 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)

  3. A Systematic Review of Culturally Specific Interventions to Increase Physical Activity for Older Asian Americans.

    Science.gov (United States)

    Katigbak, Carina; Flaherty, Erin; Chao, Ying-Yu; Nguyen, Tam; Cheung, Daphne; Yiu-Cho Kwan, Rick

    Physical activity (PA) is a significant modifiable risk factor for cardiovascular disease. For older adults, engaging in PA is shown to improve cardiac status, reduce cognitive, and functional decline, and improve overall quality of life. However, only 17% of Asian American adults meet the 2008 federal recommended guidelines for aerobic and muscle strengthening activity; and there is a paucity of data reporting on older Asian Americans - a rapidly growing, underserved group. While data pertaining to Asian Americans is frequently reported at the aggregate level, this masks differences (eg, language, culture, income) among Asian ethnic subgroups that may impact health behaviors. The purpose of this review was to identify intervention, and cultural adaptation strategies in studies promoting PA for older Asian Americans. A comprehensive literature search was performed to identify interventions published between 1996-2016 focused on improving PA among older Asian Americans (> 60 years old). Data were abstracted to examine intervention study designs, cultural adaptation strategies, theoretical frameworks, and physical activity measures. Nine studies met the review's inclusion criteria. Community-based recruitment approaches were widely used, and all studies employed cultural adaptation to varying degrees. Most studies reported improvements in PA outcomes, focused on Chinese Americans, and relied on self-reports of PA, while few aimed to increase PA using a multi-component approach. Future studies would benefit from larger sample sizes, a wider representation of Asian ethnic subgroups, and concentrated efforts to implement deep level adaptations that may increase the salience and sustainability of these interventions.

  4. 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.

  5. Use of Selective Fungal Culture Media Increases Rates of Detection of Fungi in the Respiratory Tract of Cystic Fibrosis Patients.

    Science.gov (United States)

    Hong, Gina; Miller, Heather B; Allgood, Sarah; Lee, Richard; Lechtzin, Noah; Zhang, Sean X

    2017-04-01

    The prevalence of fungi in the respiratory tracts of cystic fibrosis (CF) patients has risen. However, fungal surveillance is not routinely performed in most clinical centers in the United States, which may lead to an underestimation of the true prevalence of the problem. We conducted a prospective study comparing the rates of detection for clinically important fungi (CIF), defined as Aspergillus , Scedosporium , and Trichosporon species and Exophiala dermatitidis , in CF sputa using standard bacterial and selective fungal culture media, including Sabouraud dextrose agar with gentamicin (SDA), inhibitory mold agar (IMA), and brain heart infusion (BHI) agar with chloramphenicol and gentamicin. We described the prevalence of these fungi in an adult CF population. A total of 487 CF respiratory samples were collected from 211 unique participants. CIF were detected in 184 (37.8%) samples. Only 26.1% of CIF-positive samples were detected in bacterial culture medium, whereas greater rates of detection for fungi were found in IMA (65.8%; P culture media and longer incubation periods yielded higher rates of detection for CIF in CF sputum samples compared with that detected in bacterial culture medium, resulting in an underdetection of fungi by bacterial culture alone. The prevalence of fungi in CF may be better estimated by using selective fungal culture media, and this may translate to important clinical decisions. Copyright © 2017 American Society for Microbiology.

  6. Marketing to increase participation in a Web-based continuing medical education cultural competence curriculum.

    Science.gov (United States)

    Estrada, Carlos A; Krishnamoorthy, Periyakaruppan; Smith, Ann; Staton, Lisa; Korf, Michele J; Allison, Jeroan J; Houston, Thomas K

    2011-01-01

    CME providers may be interested in identifying effective marketing strategies to direct users to specific content. Online advertisements for recruiting participants into activities such as clinical trials, public health programs, and continuing medical education (CME) have been effective in some but not all studies. The purpose of this study was to compare the impact of 2 marketing strategies in the context of an online CME cultural competence curriculum (www.c-comp.org). In an interrupted time-series quasi-experimental design, 2 marketing strategies were tested: (1) wide dissemination to relevant organizations over a period of approximately 4 months, and (2) Internet paid search using Google Ads (5 consecutive 8-week periods--control 1, cultural/CME advertisement, control 2, hypertension/ content advertisement, control 3). Outcome measures were CME credit requests, Web traffic (visits per day, page views, pages viewed per visit), and cost. Overall, the site was visited 19,156 times and 78,160 pages were viewed. During the wide dissemination phase, the proportion of visits requesting CME credit decreased between the first (5.3%) and second (3.3%) halves of this phase (p = .04). During the Internet paid search phase, the proportion of visits requesting CME credit was highest during the cultural/CME advertisement period (control 1, 1.4%; cultural/CME ad, 4.3%; control 2, 1.5%; hypertension/content ad, 0.6%; control 3, 0.8%; p advertisement periods. The incremental cost for the cultural advertisement per CME credit requested was US $0.64. Internet advertisement focusing on cultural competence and CME was associated with about a threefold increase in requests for CME credit at an incremental cost of under US $1; however, Web traffic changes were independent of the advertisement strategy. Copyright © 2011 The Alliance for Continuing Medical Education, the Society for Academic Continuing Medical Education, and the Council on CME, Association for Hospital Medical

  7. 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…

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. Increasing Mathematics and Science Achievement for Culturally Diverse Students through Teaching Training

    Science.gov (United States)

    Mahon, Lee

    1997-01-01

    The purpose of this proposal was to field test and evaluate a Teacher Training program that would prepare teachers to increase the motivation and achievement of culturally diverse students in the areas of science and mathematics. Designed as a three year program, this report covers the first two years of the training program at the Ronald McNair School in the Ravenswood School district, using the resources of the NASA Ames Research Center and the California Framework for Mathematics and Science.

  16. Effectiveness of culturally focused interventions in increasing the satisfaction of hospitalized Asian patients: a systematic review.

    Science.gov (United States)

    Alfred, Millicent; Ubogaya, Karolina; Chen, Xing; Wint, Diana; Worral, Priscilla Sandford

    2016-08-01

    Patient satisfaction is a driving force for healthcare organizations to enhance patient services. As the Asian population in the United States is increasing at a significant rate, it is important to understand the needs of this population to implement culturally focused services that will lead to increased Asian in-patient satisfaction. The objective of the systematic review was to identify the best available evidence on the effectiveness of culturally focused interventions in increasing satisfaction of hospitalized adult Asian patients. This review considered studies that included Asian adults, 18 years of age and older, who were admitted to acute-care hospitals in countries where Asians are a minority culture. This review considered studies that included any intervention or sets of interventions implemented by hospitals for the purpose of making the hospital experience consistent with the cultural preferences of adult Asian in-patients. Satisfaction of adult Asian hospitalized patients as measured by self-report satisfaction scales or tools considered by accrediting and/or governing bodies to be acceptable sources of evidence of patients' perceptions of their care. This review first considered randomized controlled trials (RCTs), non-RCTs and quasi-experimental studies. As no RCTs or quasi-experimental studies were found, the reviewers also considered before and after studies, cohort studies and case-control studies for inclusion. This review also considered for inclusion descriptive study designs including case series, individual case reports and descriptive cross-sectional studies related to the adult Asian population in acute-care hospital settings. Three descriptive studies were selected in the review. The search strategy aimed to find both published and unpublished studies in English and Chinese (Mandarin and Cantonese) languages. A search of MEDLINE, Cumulative Index to Nursing and Allied Health Literature (CINAHL), PsycINFO, Educational Research

  17. Proteome analysis of Aspergillus niger: Lactate added in starch-containing medium can increase production of the mycotoxin fumonisin B2 by modifying acetyl-CoA metabolism

    DEFF Research Database (Denmark)

    Sørensen, Louise Marie; Lametsch, Rene; Andersen, Mikael Rørdam

    2009-01-01

    Background Aspergillus niger is a filamentous fungus found in the environment, on foods and feeds and is used as host for production of organic acids, enzymes and proteins. The mycotoxin fumonisin B2 was recently found to be produced by A. niger and hence very little is known about production...... and regulation of this metabolite. Proteome analysis was used with the purpose to reveal how fumonisin B2 production by A. niger is influenced by starch and lactate in the medium. Results Fumonisin B2 production by A. niger was significantly increased when lactate and starch were combined in the medium....... Production of a few other A. niger secondary metabolites was affected similarly by lactate and starch (fumonisin B4, orlandin, desmethylkotanin and pyranonigrin A), while production of others was not (ochratoxin A, ochratoxin alpha, malformin A, malformin C, kotanin, aurasperone B and tensidol B...

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. Increased sensitivity to ET-1 in rat cerebral arteries following organ culture

    DEFF Research Database (Denmark)

    Hansen-Schwartz, J; Edvinsson, L

    2000-01-01

    Endothelin-1 (ET-1) is recognized as being involved in the pathophysiology of cerebrovascular diseases. Using organ culture as a model for possible pathological changes we studied changes in ET(A) and ETB receptor function using a sensitive in vitro method. We observed an up-regulation of the ET......(B) receptor and an amazingly increased sensitivity to ET-1 by 3 log units in pEC50; pEC50(fresh) was 8.7 +/- 0.1, and pEC50(cultured) was 11.7 +/- 0.3. pA2 for FR139317 in the fresh vessel was 7.0 +/- 0.2 whereas it could not be obtained for the cultured vessel, indicating a possible cross-talk between the ET......(A) and ET(B) receptors. The increased sensitivity to ET-1 could also take place during cerebrovascular disease such as stroke or haemorrhage rendering the vessels considerably more sensitive to ET-1....

  3. Diabetes increases susceptibility of primary cultures of rat proximal tubular cells to chemically induced injury

    International Nuclear Information System (INIS)

    Zhong Qing; Terlecky, Stanley R.; Lash, Lawrence H.

    2009-01-01

    Diabetic nephropathy is characterized by increased oxidative stress and mitochondrial dysfunction. In the present study, we prepared primary cultures of proximal tubular (PT) cells from diabetic rats 30 days after an ip injection of streptozotocin and compared their susceptibility to oxidants (tert-butyl hydroperoxide, methyl vinyl ketone) and a mitochondrial toxicant (antimycin A) with that of PT cells isolated from age-matched control rats, to test the hypothesis that PT cells from diabetic rats exhibit more cellular and mitochondrial injury than those from control rats when exposed to these toxicants. PT cells from diabetic rats exhibited higher basal levels of reactive oxygen species (ROS) and higher mitochondrial membrane potential, demonstrating that the PT cells maintain the diabetic phenotype in primary culture. Incubation with either the oxidants or mitochondrial toxicant resulted in greater necrotic and apoptotic cell death, greater evidence of morphological damage, greater increases in ROS, and greater decreases in mitochondrial membrane potential in PT cells from diabetic rats than in those from control rats. Pretreatment with either the antioxidant N-acetyl-L-cysteine or a catalase mimetic provided equivalent protection of PT cells from both diabetic and control rats. Despite the greater susceptibility to oxidative and mitochondrial injury, both cytoplasmic and mitochondrial glutathione concentrations were markedly higher in PT cells from diabetic rats, suggesting an upregulation of antioxidant processes in diabetic kidney. These results support the hypothesis that primary cultures of PT cells from diabetic rats are a valid model in which to study renal cellular function in the diabetic state.

  4. Increased methane production in cyanobacteria and methanogenic microbe co-cultures.

    Science.gov (United States)

    Yeung, Tracey; Kwan, Matthew; Adler, Lewis; Mills, Toby J; Neilan, Brett A; Conibeer, Gavin; Patterson, Robert

    2017-11-01

    A novel light-to-bioenergy system produced 3.5 times the baseline methane output using a co-culture of cyanobacteria (Oscillatoria sp.) and a methanogenic microbial community. Analysis of micronutrients in the system during the growth phase indicated that cobalt, iron, nickel and zinc were not appreciably consumed. The stable consumption and return of macronutrients calcium and magnesium were also observed. Essential macronutrients nitrogen, in the form of nitrate, and phosphorus showed no cycling during the growth phase and were depleted at rates of 0.35mg/L/day and 0.40µg/L/day, respectively. Biofilm formation increased the resilience of biomass to bacterial degradation in an anaerobic digester, as shown by viability assays of cyanobacterial biofilms in the co-culture. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Photosynthetic light reactions increase total lipid accumulation in carbon-supplemented batch cultures of Chlorella vulgaris.

    Science.gov (United States)

    Woodworth, Benjamin D; Mead, Rebecca L; Nichols, Courtney N; Kolling, Derrick R J

    2015-03-01

    Microalgae are an attractive biofuel feedstock because of their high lipid to biomass ratios, lipid compositions that are suitable for biodiesel production, and the ability to grow on varied carbon sources. While algae can grow autotrophically, supplying an exogenous carbon source can increase growth rates and allow heterotrophic growth in the absence of light. Time course analyses of dextrose-supplemented Chlorella vulgaris batch cultures demonstrate that light availability directly influences growth rate, chlorophyll production, and total lipid accumulation. Parallel photomixotrophic and heterotrophic cultures grown to stationary phase reached the same amount of biomass, but total lipid content was higher for algae grown in the presence of light (an average of 1.90 mg/mL vs. 0.77 mg/mL over 5 days of stationary phase growth). Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. 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.

  7. Increased lipids production of Nannochloropsis oculata and Chlorella vulgaris for biodiesel synthesis through the optimization of growth medium composition arrangement by using bicarbonate addition

    Directory of Open Access Journals (Sweden)

    Dianursanti

    2018-01-01

    Full Text Available Chlorella vulgaris and Nannochloropsis oculata are a highly potential microalgae to be used in pilot-scale of biodiesel synthesis. The essential content from these microalgae is the fatty acid of lipid which is the main target for the feed and biodiesel industries. One of the key factor in improving lipid microalgae are the arrangemment of nutrients in the growth medium. Research on the regulation of nutrients using bicarbonate (HCO3- as an additional inorganic carbon source has been done by many studies, but the yield of lipids obtained has not been much. The aim of the study was to improve the lipid yield of Chlorella vulgaris and Nannochloropsis oculata. Variation of [HCO3-] which added to Walne medium were 25 ppm and 75 ppm, while the Walne medium without the addition of bicarbonate acts as control. The results showed that [HCO3-] 75 ppm could increase Chlorella vulgaris biomass by 0.9162 g/l with 17.0% wt, while Nannochloropsis oculata produced the greatest lipid content in [HCO3-] 25 ppm of 20.3% wt and the largest biomass on [HCO3-] 75 ppm of 1.7233 g/l.

  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. 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

  10. 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.

  11. 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.

  12. 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.

  13. Cultural stereotypes as gatekeepers: increasing girls’ interest in computer science and engineering by diversifying stereotypes

    Science.gov (United States)

    Cheryan, Sapna; Master, Allison; Meltzoff, Andrew N.

    2015-01-01

    Despite having made significant inroads into many traditionally male-dominated fields (e.g., biology, chemistry), women continue to be underrepresented in computer science and engineering. We propose that students’ stereotypes about the culture of these fields—including the kind of people, the work involved, and the values of the field—steer girls away from choosing to enter them. Computer science and engineering are stereotyped in modern American culture as male-oriented fields that involve social isolation, an intense focus on machinery, and inborn brilliance. These stereotypes are compatible with qualities that are typically more valued in men than women in American culture. As a result, when computer science and engineering stereotypes are salient, girls report less interest in these fields than their male peers. However, altering these stereotypes—by broadening the representation of the people who do this work, the work itself, and the environments in which it occurs—significantly increases girls’ sense of belonging and interest in the field. Academic stereotypes thus serve as gatekeepers, driving girls away from certain fields and constraining their learning opportunities and career aspirations. PMID:25717308

  14. Cultural stereotypes as gatekeepers: increasing girls' interest in computer science and engineering by diversifying stereotypes.

    Science.gov (United States)

    Cheryan, Sapna; Master, Allison; Meltzoff, Andrew N

    2015-01-01

    Despite having made significant inroads into many traditionally male-dominated fields (e.g., biology, chemistry), women continue to be underrepresented in computer science and engineering. We propose that students' stereotypes about the culture of these fields-including the kind of people, the work involved, and the values of the field-steer girls away from choosing to enter them. Computer science and engineering are stereotyped in modern American culture as male-oriented fields that involve social isolation, an intense focus on machinery, and inborn brilliance. These stereotypes are compatible with qualities that are typically more valued in men than women in American culture. As a result, when computer science and engineering stereotypes are salient, girls report less interest in these fields than their male peers. However, altering these stereotypes-by broadening the representation of the people who do this work, the work itself, and the environments in which it occurs-significantly increases girls' sense of belonging and interest in the field. Academic stereotypes thus serve as gatekeepers, driving girls away from certain fields and constraining their learning opportunities and career aspirations.

  15. Proteome analysis of Aspergillus niger: Lactate added in starch-containing medium can increase production of the mycotoxin fumonisin B2 by modifying acetyl-CoA metabolism

    DEFF Research Database (Denmark)

    Sørensen, Louise Marie; Lametsch, Rene; Andersen, Mikael Rørdam

    2009-01-01

    containing nitrate and starch can increase fumonisin B2 production by A. niger as well as production of some other secondary metabolites. Changes in the balance of intracellular metabolites towards a higher level of carbon passing through acetyl-CoA and a high capacity to regenerate NADPH during growth...... on medium with starch and lactate were found to be the likely cause of this effect. The results lead to the hypothesis that fumonisin production by A. niger is regulated by acetyl-CoA....

  16. Device for selective culturing of bacteria used to increase oil bed output

    Energy Technology Data Exchange (ETDEWEB)

    Lazar, J; Zamfirescu, I

    1982-12-15

    The device is patented for selective culturing of bacteria which can be used for increasing oil bed output in order to adapt them to conditions of the field. Development and reproduction of the bacteria are recorded by increase in pressure indicated by the manometer. When the pressure in the container reaches 0.5-0.5 MPa, the gases formed in it are released into the atmosphere through the reverse valve and the hose. Slow rise in pressure in the vessel (usually within 5-10 days) indicates that the nutrient carrier is approaching depletion. After removal of the gases formed because of bacterial activity, the cover is opened and a sample is taken for microbiological analysis and establishment of bacterial growth required for increasing oil output in the given field. When it is established that the developing populations of bacteria correspond to the requirements, then the necessary quantity of bacteria are grown under the same conditions in a battery of similar devices.

  17. RELATIONSHIP BETWEEN THE EFFECTS OF INCREASING THE SATISFACTION OF THE VISITORS AND DEVELOPMENT OF CULTURAL TOURISM

    Directory of Open Access Journals (Sweden)

    Tünde SZABÓ

    2011-12-01

    Full Text Available In order to succeed on the market, organizations must permanently focus their efforts towards understanding and fulfilling the necessities and expectations of all their clients, both the current and the potential ones. At the same time, enterprises must identify methods and tools appropriate for evaluating the satisfaction degree of the clients and for ensuring the loyalty of the most important clients, as well as improving the internal and external relational system and creating new partnerships with loyal clients. The effects of increasing the client satisfaction can be numerous, and they depend greatly on the activities specific to each organization. In the current paper, an attempt is made to highlight the possible effects of increasing the satisfaction of the visitors of the National Seckler Museum from Sfîntu Gheorghe, - effects like an increased number of visitors, tourist attraction, developing and promoting cultural tourism in the county.

  18. [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.

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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+).

  4. 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.

  5. 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.

  6. 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.

  7. Design and deployment strategies for small and medium sized reactors (SMRs) to overcome loss of economies of scale and incorporate increased proliferation resistance

    International Nuclear Information System (INIS)

    Kuznetsov, V.

    2007-01-01

    The designers of innovative small and medium sized reactors pursue new design and deployment strategies making use of certain advantages provided by smaller reactor size and capacity to achieve reduced design complexity and simplified operation and maintenance requirements, and to provide for incremental capacity increase through multi-module plant clustering. Competitiveness of SMRs (Small and Medium size Reactor) depends on the incorporated strategies to overcome loss of economies of scale but equally it depends on finding appropriate market niches for such reactors. For many less developed countries, these are the features of enhanced proliferation resistance and increased robustness of barriers for sabotage protection that may ensure the progress of nuclear power. For such countries, small reactors without on-site refuelling, designed for infrequent replacement of well-contained fuel cassette(s) in a manner that impedes clandestine diversion of nuclear fuel material, may provide a solution. Based on the outputs of recent IAEA activities for innovative SMRs, the paper provides a summary of the state-of-the-art in approaches to improve SMR competitiveness and incorporate enhanced proliferation resistance and energy security. (author)

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  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. Increased production of azadirachtin from an improved method of androgenic cultures of a medicinal tree Azadirachta indica A. Juss.

    Science.gov (United States)

    Srivastava, Priyanka; Chaturvedi, Rakhi

    2011-07-01

    Present report is the first direct evidence of azadirachtin production in androgenic haploid cultures of Azadirachta indica, a woody medicinal tree. Anther cultures at early-late-uninucleate stage of microspores were established on MS medium with BAP (5 μM), 2,4-D (1 μM) and NAA (1 μM) containing 12% sucrose. The calli, induced, were further multiplied on 2,4-D and Kinetin media. Shoots, differentiated on BAP (2.2 μM) + NAA (0.05 μM) medium, were elongated on MS + BAP (0.5 μM) and multiplied on MS + BAP (1 μM) + CH (250 mg/l). Thereafter, the shoots were rooted on ¼ MS + IBA (0.5 μM). Cytological analysis of the calli and regenerants have confirmed their haploid status with the chromosome number as 2n = x = 12. The haploid cell lines and leaves from in vitro grown plantlets were analyzed for azadirachtin by RP-HPLC and mass spectroscopy. Maximum azadirachtin (728.41 μg/g DW) was detected in calli supporting best shoot proliferation while least (49 μg/g DW) was observed in an undifferentiated line from maintenance medium. This study has brought us a step closer to develop genetically pure lines that could serve as new and attractive alternative ways of homogeneous controlled production of high value compounds, round the year, independent of geographical and climatic barrier.

  14. Bioaugmentation of Syntrophic Acetate-Oxidizing Culture in Biogas Reactors Exposed to Increasing Levels of Ammonia

    Science.gov (United States)

    Westerholm, Maria; Levén, Lotta

    2012-01-01

    The importance of syntrophic acetate oxidation for process stability in methanogenic systems operating at high ammonia concentrations has previously been emphasized. In this study we investigated bioaugmentation of syntrophic acetate-oxidizing (SAO) cultures as a possible method for decreasing the adaptation period of biogas reactors operating at gradually increased ammonia concentrations (1.5 to 11 g NH4+-N/liter). Whole stillage and cattle manure were codigested semicontinuously for about 460 days in four mesophilic anaerobic laboratory-scale reactors, and a fixed volume of SAO culture was added daily to two of the reactors. Reactor performance was evaluated in terms of biogas productivity, methane content, pH, alkalinity, and volatile fatty acid (VFA) content. The decomposition pathway of acetate was analyzed by isotopic tracer experiments, and population dynamics were monitored by quantitative PCR analyses. A shift in dominance from aceticlastic methanogenesis to SAO occurred simultaneously in all reactors, indicating no influence by bioaugmentation on the prevailing pathway. Higher abundances of Clostridium ultunense and Tepidanaerobacter acetatoxydans were associated with bioaugmentation, but no influence on Syntrophaceticus schinkii or the methanogenic population was distinguished. Overloading or accumulation of VFA did not cause notable dynamic effects on the population. Instead, the ammonia concentration had a substantial impact on the abundance level of the microorganisms surveyed. The addition of SAO culture did not affect process performance or stability against ammonia inhibition, and all four reactors deteriorated at high ammonia concentrations. Consequently, these findings further demonstrate the strong influence of ammonia on the methane-producing consortia and on the representative methanization pathway in mesophilic biogas reactors. PMID:22923397

  15. 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.

  16. Medium doses of daily vitamin D decrease falls and higher doses of daily vitamin D3 increase falls: A randomized clinical trial.

    Science.gov (United States)

    Smith, Lynette M; Gallagher, J Christopher; Suiter, Corinna

    2017-10-01

    Falls are a serious health problem in the aging population. Because low levels of vitamin D have been associated with increased fall rates, many trials have been performed with vitamin D; two meta-analyses showed either a small effect or no effect of vitamin D on falls. We conducted a study of the effect of vitamin D on serum 25 hydroxyvitamin D (25OHD) and data on falls was collected as a secondary outcome. In a 12-month double blind randomized placebo trial, elderly women, mean age 66 years, were randomized to one of seven daily oral doses of vitamin D or placebo. The main inclusion criterion for study was a baseline serum 25OHDvitamin D on falls followed a U-shaped curve whether analyzed by dose or serum 25OHD levels. There was no decrease in falls on low vitamin D doses 400, 800 IU, a significant decrease on medium doses 1600, 2400,3200 IU (p=0.020) and no decrease on high doses 4000, 4800 IU compared to placebo (p=0.55). When compared to 12-month serum 25OHD quintiles, the faller rate was 60% in the lowest quintile <25ng/ml (<50nmol/L), 21% in the low middle quintile 32-38ng/ml (80-95nmo/L), 72% in the high middle quintile 38-46ng/ml (95-115nmo/L) and 45% in the highest quintile 46-66ng/ml (115-165nmol/L). In the subgroup with a fall history, fall rates were 68% on low dose, 27% on medium doses and 100% on higher doses. Fall rates on high doses were increased compared to medium doses (Odds Ratio 5.6.95% CI: 2.1-14.8). In summary, the maximum decrease in falls corresponds to a 12- month serum 25OHD of 32-38ng/ml (80-95nmol/L) and faller rates increase as serum 25OHD exceed 40-45ng/ml (100-112.5nmol/L). The Tolerable upper limit (TUL) recently increased in 2010 from 2000 to 4000 IU/day may need to be reduced in elderly women especially in those with a fall history. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Adaptive response of Saccharomyces cerevisiae UEME3 to isoproturon depends on the presence of peptone in the rich culture medium

    OpenAIRE

    Candeias, M; Alves-Pereira, I; Ferreira, R

    2010-01-01

    The isoproturon, a herbicide which acts as a photosynthesis inhibitor is consider by some authors as toxic to humans having ability for tumors induction. When this phenylurea is metabolized by biotransformation systems can generate oxidative stress, activating electron transport chains present at mitochondria and endoplasmic reticulum and causing an increase of intracellular ROS. Having in account exposure above, the main purpose of this work was to compare the effects of is...

  18. 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

  19. Cultural Stereotypes as Gatekeepers: Increasing Girls’ Interest in Computer Science and Engineering by Diversifying Stereotypes

    Directory of Open Access Journals (Sweden)

    Sapna eCheryan

    2015-02-01

    Full Text Available Despite having made significant inroads into many traditionally male-dominated fields (e.g., biology, chemistry, women continue to be underrepresented in computer science and engineering. We propose that students’ stereotypes about the culture of these fields—including the kind of people, the work involved, and the values of the field—steer girls away from choosing to enter these fields. Computer science and engineering are stereotyped in modern American culture as male-oriented fields that involve social isolation, an intense focus on machinery, and inborn brilliance. These stereotypes are more compatible with qualities that are typically valued in men than women. As a result, when computer science and engineering stereotypes are salient, girls report less interest in these fields than their male peers. However, altering these stereotypes—by broadening the representation of the people who do this work, the work itself, and the environments in which it occurs—significantly increases girls’ sense of belonging and interest in the field. Academic stereotypes thus serve as gatekeepers, driving girls away from certain fields and constraining their learning opportunities and career aspirations.

  20. Increased adsorption of histidine-tagged proteins onto tissue culture polystyrene.

    Science.gov (United States)

    Holmberg, Maria; Hansen, Thomas Steen; Lind, Johan Ulrik; Hjortø, Gertrud Malene

    2012-04-01

    In this study we compare histidine-tagged and native proteins with regards to adsorption properties. We observe significantly increased adsorption of proteins with an incorporated polyhistidine amino acid motif (HIS-tag) onto tissue culture polystyrene (TCPS) compared to similar proteins without a HIS-tag. The effect is not observed on polystyrene (PS). Adsorption experiments have been performed at physiological pH (7.4) and the effect was only observed for the investigated proteins that have pI values below or around 7.4. Competitive adsorption experiments with imidazole and ethylenediaminetetraacetic acid (EDTA), as well as adsorption performed at different pH and ionic strength indicates that the high adsorption is caused by electrostatic interaction between negatively charged carboxylate groups on the TCPS surface and positively charged histidine residues in the proteins. Pre-adsorption of bovine serum albumin (BSA) does not decrease the adsorption of HIS-tagged proteins onto TCPS. Our findings identify a potential problem in using HIS-tagged signalling molecule in assays with cells cultured on TCPS, since the concentration of the molecule in solution might be affected and this could critically influence the assay outcome. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. Eficacia del medio de cultivo CromoCen(R CC en el diagnóstico de la infección urinaria Efficacy of the CromoCen (R culture medium in the diagnosis of urinary infection

    Directory of Open Access Journals (Sweden)

    Emilio Simón Barroso de la Cruz

    2006-12-01

    Full Text Available Se realizó la evaluación del desempeño con muestras clínicas de orina para determinar la eficacia del CromoCen(R CC como medio de cultivo en el diagnóstico de la infección urinaria, e identificar, con su utilización, los grupos bacterianos más frecuentes y el tiempo necesario para realizar el diagnóstico. En una muestra de 500 casos atendidos en la consulta externa de urología y realizados en el servicio de microbiología de San Antonio de los Baños, en la provincia de La Habana, en el período comprendido desde enero a junio de 2003. Se confirmó la hipótesis de trabajo: el medio de cultivo estudiado fue eficaz en el diagnóstico rápido de la infección urinaria. Desde el punto de vista ético se mantuvo la confidencialidad, se elevó la autoestima del binomio médico-paciente, y su calidad de vida desde el punto de vista bioético, por lo que se recomienda emplear este medio de cultivo en el diagnóstico rápido de la infección urinaria.The evaluation of the performance with clinical samples of urine was carried out to determine the efficacy of CromoCen(R CC as a culture medium in the diagnosis of urinary infection, and to identify with its use the most frequent bacterial groups and the time necessary to make the diagnosis. In a sample of 500 cases seen at the aoutpatient department of urology and analyzed at the microbiology service of San Antonio de los Baños, in the province of Havana, from January to June, 2003, it was confirmed the working hypothesis: the studied culture medium was efficient for the rapid diagnosis of urinary infection. From the ethical point of view, the reliability was maintained, and the self-steem of the physician-patient binomial, as well as the quality of life of the patient from the bioethical view increased. That's why, it is recommended to use this culture medium in the rapid diagnosis of the urinary infection.

  2. Evaluation of entomopathogenic nematodes and the supernatants of the in vitro culture medium of their mutualistic bacteria for the control of the root-knot nematodes Meloidogyne incognita and M. arenaria.

    Science.gov (United States)

    Kepenekci, Ilker; Hazir, Selcuk; Lewis, Edwin E

    2016-02-01

    The suppressive effects of various formulations of four entomopathogenic nematode (EPN) species and the supernatants of their mutualistic bacteria on the root-knot nematodes (RKNs) Meloidogyne incognita and M. arenaria in tomato roots were evaluated. The EPNs Steinernema carpocapsae, S. feltiae, S. glaseri and Heterorhabditis bacteriophora were applied as either live infective juveniles (IJs) or infected insect cadavers. Spent medium from culturing the bacterial symbionts Xenorhabdus bovienii and Photorhabdus luminescens kayaii with the cells removed was also applied without their nematode partners. The aqueous suspensions of IJs, infected cadaver applications of EPNs and especially treatments of X. bovienii supernatant suppressed the negative impact of RKNs on tomatoes. Specific responses to treatment were reduced RKN egg masses, increased plant height and increased fresh and dry weights compared with the control where only RKNs were applied. Among the treatments tested, the plant-dipping method of X. bovienii into bacterial culture fluid may be the most practical and effective method for M. incognita and M. arenaria control. © 2015 Society of Chemical Industry.

  3. Supplementing five-point body condition score with body fat percentage increases the sensitivity for assessing overweight status of small to medium sized dogs

    Directory of Open Access Journals (Sweden)

    Arai T

    2012-09-01

    Full Text Available Gebin Li,1 Peter Lee,1 Nobuko Mori,1 Ichiro Yamamoto,1 Koh Kawasumi,1 Hisao Tanabe,2 Toshiro Arai11Department of Veterinary Science, School of Veterinary Medicine, Nippon Veterinary and Life Science University, 2Komazawa Animal Hospital, Tokyo, JapanBackground and methods: Currently, five-point body condition scoring (BCS is widely used by veterinarians and clinicians to assess adiposity in dogs in Japan. However, BCS score assignment is subjective in nature, and most clinicians do not score with half points, instead preferring to round off values, thereby rendering less accurate assessments. Therefore, we sought to determine whether assessing body fat percentage using simple morphometric measurements and supplementing this with five-point BCS can have increased sensitivity for detecting increasing adiposity in overweight small-medium sized dog breeds via plasma metabolite validation.Results: Overall, lean body fat percentage was determined to be 15%–22% for male (non-neutered/neutered dogs and 15%–25% for female (nonspayed/spayed. Dogs categorized as overweight by BCS had significantly higher levels of nonesterified fatty acids (P = 0.005, whereas animals categorized as overweight by BCS + body fat percentage were observed to have significantly higher levels of nonesterified fatty acids (P = 0.006, total cholesterol (P = 0.029, and triglycerides (P = 0.001 than lean animals. The increased sensitivity due to body fat percentage for gauging alterations in plasma metabolite levels may be due to increased correlation strength. Body fat percentage correlated positively with plasma insulin (r = 0.627, P = 0.002, nonesterified fatty acids (r = 0.674, P < 0.001, total cholesterol (r = 0.825, P < 0.0001, triglycerides (r = 0.5823, P < 0.005, blood urea nitrogen (r = 0.429, P < 0.05, creatinine (r = 0.490, P = 0.021, and total protein (r = 0.737, P< 0.0001 levels, which all tend to increase as a result of increasing adiposity

  4. Increased adsorption of histidine-tagged proteins onto tissue culture polystyrene

    DEFF Research Database (Denmark)

    Holmberg, Maria; Hansen, Thomas Steen; Lind, Johan Ulrik

    2012-01-01

    and ethylenediaminetetraacetic acid (EDTA), as well as adsorption performed at different pH and ionic strength indicates that the high adsorption is caused by electrostatic interaction between negatively charged carboxylate groups on the TCPS surface and positively charged histidine residues in the proteins. Pre......In this study we compare histidine-tagged and native proteins with regards to adsorption properties. We observe significantly increased adsorption of proteins with an incorporated polyhistidine amino acid motif (HIS-tag) onto tissue culture polystyrene (TCPS) compared to similar proteins without...... a HIS-tag. The effect is not observed on polystyrene (PS). Adsorption experiments have been performed at physiological pH (7.4) and the effect was only observed for the investigated proteins that have pI values below or around 7.4. Competitive adsorption experiments with imidazole...

  5. Corporate Culture- the Way of Increasing the Work Quality in Transport Organization

    Directory of Open Access Journals (Sweden)

    Alžbeta Bielikova

    2008-01-01

    Full Text Available The article deals with corporate culture in transport organization.The corporate culture has a momentary character and itis affected by continual confrontation with business prioritiesand by movement of external environment. For this reason theactivity of transport organization must be still adapted to customerexpectations and needs and c01porate culture must beadapted to this objective.

  6. Low Fat Loss Response after Medium-Term Supervised Exercise in Obese Is Associated with Exercise-Induced Increase in Food Reward

    Directory of Open Access Journals (Sweden)

    Graham Finlayson

    2011-01-01

    Full Text Available Objective. To examine exercise-induced changes in the reward value of food during medium-term supervised exercise in obese individuals. Subjects/Methods. The study was a 12-week supervised exercise intervention prescribed to expend 500 kcal/day, 5 d/week. 34 sedentary obese males and females were identified as responders (R or non-responders (NR to the intervention according to changes in body composition relative to measured energy expended during exercise. Food reward (ratings of liking and wanting, and relative preference by forced choice pairs for an array of food images was assessed before and after an acute exercise bout. Results. 20 responders and 14 non-responders were identified. R lost 5.2 kg ± 2.4 of total fat mass and NR lost 1.7 kg ± 1.4. After acute exercise, liking for all foods increased in NR compared to no change in R. Furthermore, NR showed an increase in wanting and relative preference for high-fat sweet foods. These differences were independent of 12-weeks regular exercise and weight loss. Conclusion. Individuals who showed an immediate post-exercise increase in liking and increased wanting and preference for high-fat sweet foods displayed a smaller reduction in fat mass with exercise. For some individuals, exercise increases the reward value of food and diminishes the impact of exercise on fat loss.

  7. High-yield production of biologically active recombinant protein in shake flask culture by combination of enzyme-based glucose delivery and increased oxygen transfer

    Directory of Open Access Journals (Sweden)

    Ukkonen Kaisa

    2011-12-01

    Full Text Available Abstract This report describes the combined use of an enzyme-based glucose release system (EnBase® and high-aeration shake flask (Ultra Yield Flask™. The benefit of this combination is demonstrated by over 100-fold improvement in the active yield of recombinant alcohol dehydrogenase expressed in E. coli. Compared to Terrific Broth and ZYM-5052 autoinduction medium, the EnBase system improved yield mainly through increased productivity per cell. Four-fold increase in oxygen transfer by the Ultra Yield Flask contributed to higher cell density with EnBase but not with the other tested media, and consequently the product yield per ml of EnBase culture was further improved.

  8. Increased detection of mastitis pathogens by real-time PCR compared to bacterial culture.

    Science.gov (United States)

    Keane, O M; Budd, K E; Flynn, J; McCoy, F

    2013-09-21

    Rapid and accurate identification of mastitis pathogens is important for disease control. Bacterial culture and isolate identification is considered the gold standard in mastitis diagnosis but is time consuming and results in many culture-negative samples. Identification of mastitis pathogens by PCR has been proposed as a fast and sensitive alternative to bacterial culture. The results of bacterial culture and PCR for the identification of the aetiological agent of clinical mastitis were compared. The pathogen identified by traditional culture methods was also detected by PCR in 98 per cent of cases indicating good agreement between the positive results of bacterial culture and PCR. A mastitis pathogen could not be recovered from approximately 30 per cent of samples by bacterial culture, however, an aetiological agent was identified by PCR in 79 per cent of these samples. Therefore, a mastitis pathogen was detected in significantly more milk samples by PCR than by bacterial culture (92 per cent and 70 per cent, respectively) although the clinical relevance of PCR-positive culture-negative results remains controversial. A mixed infection of two or more mastitis pathogens was also detected more commonly by PCR. Culture-negative samples due to undetected Staphylococcus aureus infections were rare. The use of PCR technology may assist in rapid mastitis diagnosis, however, accurate interpretation of PCR results in the absence of bacterial culture remains problematic.

  9. Growth regulators and darkness increase efficiency in in vitro culture of immature embryos from peppers

    Directory of Open Access Journals (Sweden)

    Juan Pablo Manzur

    2014-12-01

    Full Text Available Common pepper (Capsicum annuum L. is one of the most important vegetables in the world, and extensive breeding efforts are being made to develop new improved strains of this species. In this regard, in vitro culture of immature embryos may help breeders accelerate breeding cycles and overcome interspecific barriers, among other applications. In this study, we have optimized a protocol for in vitro culture of immature embryos of C. annuum. Levels of indole-3-acetic acid (IAA and zeatin have been tested to improve the efficiency (germination rates of this technique in C. annuum embryos at the four main immature stages (i.e. globular, heart, torpedo, and early cotyledonary from four varietal types of this species (California Wonder, Piquillo, Guindilla, and Bola. The effect of 5-day initial incubation in the dark was also tested on the most efficient hormone formulation. On average, relatively low levels of both IAA and zeatin (0.01 mg L−¹ each (M1 provided the highest germination rates, particularly in the advanced stages (torpedo and cotyledonary. To a lesser extent, the lack of these growth regulators (M0 or high IAA (0.2 mg L−¹/low zeatin (0.01 mg L−¹ (M2 combination also had a positive response. On the contrary, high zeatin levels (0.2 mg L−¹ produced very low germination rates or callus development (efficiency 0-7 %. Different responses were also found between genotypes. Thus, considering the best media (M0, M1, M2, Bola embryos had the highest rates. M1 plus 5-days of initial dark incubation (M1-D improved the efficiency rates at all embryo stages, particularly in the earliest (globular embryos which increased from 3 % to > 20 %.

  10. Glycocalyx Degradation Induces a Proinflammatory Phenotype and Increased Leukocyte Adhesion in Cultured Endothelial Cells under Flow.

    Directory of Open Access Journals (Sweden)

    Karli K McDonald

    Full Text Available Leukocyte adhesion to the endothelium is an early step in the pathogenesis of atherosclerosis. Effective adhesion requires the binding of leukocytes to their cognate receptors on the surface of endothelial cells. The glycocalyx covers the surface of endothelial cells and is important in the mechanotransduction of shear stress. This study aimed to identify the molecular mechanisms underlying the role of the glycocalyx in leukocyte adhesion under flow. We performed experiments using 3-D cell culture models, exposing human abdominal aortic endothelial cells to steady laminar shear stress (10 dynes/cm2 for 24 hours. We found that with the enzymatic degradation of the glycocalyx, endothelial cells developed a proinflammatory phenotype when exposed to uniform steady shear stress leading to an increase in leukocyte adhesion. Our results show an up-regulation of ICAM-1 with degradation compared to non-degraded controls (3-fold increase, p<0.05 and we attribute this effect to a de-regulation in NF-κB activity in response to flow. These results suggest that the glycocalyx is not solely a physical barrier to adhesion but rather plays an important role in governing the phenotype of endothelial cells, a key determinant in leukocyte adhesion. We provide evidence for how the destabilization of this structure may be an early and defining feature in the initiation of atherosclerosis.

  11. Arginine vasopressin increases cellular free calcium concentration and adenosine 3',5'-monophosphate production in rat renal papillary collecting tubule cells in culture

    International Nuclear Information System (INIS)

    Ishikawa, S.; Okada, K.; Saito, T.

    1988-01-01

    The role of calcium (Ca) in the cellular action of arginine vasopressin (AVP) was examined in rat renal papillary collecting tubule cells in culture. AVP increased both the cellular free Ca concentration ([Ca2+]i) using fura-2, and cAMP production in a dose-dependent manner. AVP-induced cellular Ca mobilization was totally blocked by the antagonist to the antidiuretic action of AVP, and somewhat weakened by the antagonist to the vascular action of AVP. 1-Deamino-8-D-AVP (dDAVP). an antidiuretic analog of AVP, also increased [Ca2+] significantly. Cellular Ca mobilization was not obtained with cAMP, forskolin (a diterpene activator of adenylate cyclase), or phorbol-12-myristate-13-acetate. The early phase of [Ca2+]i depended on the intracellular Ca pool, since an AVP-induced rise in [Ca2+]i was obtained in cells pretreated with Ca-free medium containing 1 mM EGTA, verapamil, or cobalt, which blocked cellular Ca uptake. Also, AVP increased 45 Ca2+ influx during the initial 10 min, which initiated the sustained phase of cellular Ca mobilization. However, cellular cAMP production induced by AVP during the 10-min observation period was diminished in the cells pretreated with Ca-free medium, verapamil, or cobalt, but was still significantly higher than the basal level. This was also diminished by a high Ca concentration in medium. These results indicate that 1) AVP concomitantly regulates cellular free Ca as well as its second messenger cAMP production; 2) AVP-induced elevation of cellular free Ca is dependent on both the cellular Ca pool and extracellular Ca; and 3) there is an optimal level of extracellular Ca to modulate the AVP action in renal papillary collecting tubule cells

  12. Increasing the Thermal Conductivity of Graphene-Polyamide-6,6 Nanocomposites by Surface-Grafted Polymer Chains: Calculation with Molecular Dynamics and Effective-Medium Approximation.

    Science.gov (United States)

    Gao, Yangyang; Müller-Plathe, Florian

    2016-02-25

    By employing reverse nonequilibrium molecular dynamics simulations in a full atomistic resolution, the effect of surface-grafted chains on the thermal conductivity of graphene-polyamide-6.6 (PA) nanocomposites has been investigated. The interfacial thermal conductivity perpendicular to the graphene plane is proportional to the grafting density, while it first increases and then saturates with the grafting length. Meanwhile, the intrinsic in-plane thermal conductivity of graphene drops sharply as the grafting density increases. The maximum overall thermal conductivity of nanocomposites appears at an intermediate grafting density because of these two competing effects. The thermal conductivity of the composite parallel to the graphene plane increases with the grafting density and grafting length which is attributed to better interfacial coupling between graphene and PA. There exists an optimal balance between grafting density and grafting length to obtain the highest interfacial and parallel thermal conductivity. Two empirical formulas are suggested, which quantitatively account for the effects of grafting length and density on the interfacial and parallel thermal conductivity. Combined with effective medium approximation, for ungrafted graphene in random orientation, the model overestimates the thermal conductivity at low graphene volume fraction (f 10%). For unoriented grafted graphene, the model matches the experimental results well. In short, this work provides some valuable guides to obtain the nanocomposites with high thermal conductivity by grafting chain on the surface of graphene.

  13. Amniocar as a proliferative medium for mesenchymal cells

    Directory of Open Access Journals (Sweden)

    V. V. Chestkov

    2014-01-01

    Full Text Available Objectives. To develop the Amniocar nutrient medium that contains fetal calf serum (FCS and growth factors cocktail for mass cultivation of human fibroblasts. To study proliferative activity of the medium on cultures of HUVEC cells of mesenchymal origin and mesenchymal stromal cells, as well as on cell culture of human amniotic fluid.Materials and methods. Determination of the rate of accumulation of the cellular mass and cell morphology in the course of cultivation of cells of various histogenesis in the Amniocar medium and nutrient medium that contains 10 % of FCS.Results. It has been demonstrated that the Amniocar medium is prevalent as compared to the standard DMEM medium with 10 % of FCS by 2 to 5 times for cultivation of skin fibroblasts, HUVEC, and mesenchymal stem cells. The Amniocar medium increased the quantity of endothelial cells that enter mitosis and maintained the culture of HUVEC cells with prolonged passaging in vitro. Clonal cultivation of human amniotic fluid cells in the Amniocar medium secured development of colonies of both fibroblast and epithelial type.Conclusions. Proliferative Amniocar medium is efficient for mass cultivation of various cells of mesenchymal origin and can be used for diagnostic purposes in medical genetics, oncology, etc.

  14. [The effect of 3-aminobenzamide on the mitotic cycle of Chinese hamster cells cultured on a medium with 5-bromodeoxyuridine following ionizing radiation action].

    Science.gov (United States)

    Kirillova, T V; Rozanov, Iu M; Spivak, I M

    1992-01-01

    A specific inhibitor of poly(ADP-ribose)polymerase-3-aminobenzamide (6 mM) has been shown to: 1) reduce survival of non-irradiated CHO-K1 cells, cultivated in medium containing 5-bromodeoxyuridine (10 mkM, BDU cells), and increase their radiosensitivity; 2) induce G2 delay in BDU cells while progressing through the cell cycle as analysed by the DNA flow cytometry; 3) increase to a great degree G2 delay in X-irradiated BDU cells. 3-Aminobenzamide is primarily effective when it is present during the first or two first cell cycles after the initial addition of BDU. The above data confirm the involvement, presumably an indirect one, of ADP-ribosylation in the DNA repair through affecting the chromatin structure.

  15. 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....

  16. 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.

  17. Growth characteristics of liquid cultures increase the reliability of presumptive identification of Mycobacterium tuberculosis complex.

    Science.gov (United States)

    Pinhata, Juliana Maira Watanabe; Felippe, Isis Moreira; Gallo, Juliana Failde; Chimara, Erica; Ferrazoli, Lucilaine; de Oliveira, Rosangela Siqueira

    2018-04-23

    We evaluated the microscopic and macroscopic characteristics of mycobacteria growth indicator tube (MGIT) cultures for the presumptive identification of the Mycobacterium tuberculosis complex (MTBC) and assessed the reliability of this strategy for correctly directing isolates to drug susceptibility testing (DST) or species identification. A total of 1526 isolates of mycobacteria received at the Instituto Adolfo Lutz were prospectively subjected to presumptive identification by the observation of growth characteristics along with cord formation detection via microscopy. The presumptive identification showed a sensitivity, specificity and accuracy of 98.8, 92.5 and 97.9 %, respectively. Macroscopic analysis of MTBC isolates that would have been erroneously classified as non-tuberculous mycobacteria based solely on microscopic morphology enabled us to direct them rapidly to DST, representing a substantial gain to patients. In conclusion, the growth characteristics of mycobacteria in MGIT, when considered along with cord formation, increased the reliability of the presumptive identification, which has a great impact on the laboratory budget and turnaround times.

  18. Options for small and medium sized reactors (SMRs) to overcome loss of economies of scale and incorporate increased proliferation resistance and energy security

    International Nuclear Information System (INIS)

    Kuznetsov, Vladimir

    2008-01-01

    The designers of innovative small and medium sized reactors pursue new design and deployment strategies making use of certain advantages provided by smaller reactor size and capacity to achieve reduced design complexity and simplified operation and maintenance requirements, and to provide for incremental capacity increase through multi-module plant clustering. Competitiveness of SMRs depends on the incorporated strategies to overcome loss of economies of scale but equally it depends on finding appropriate market niches for such reactors. For many less developed countries, these are the features of enhanced proliferation resistance and increased robustness of barriers for sabotage protection that may ensure the progress of nuclear power. For such countries, small reactors without on-site refuelling, designed for infrequent replacement of well-contained fuel cassette(s) in a manner that impedes clandestine diversion of nuclear fuel material, may provide a solution. Based on the outputs of recent IAEA activities for innovative SMRs, the paper provides a summary of the state-of-the-art in approaches to improve SMR competitiveness and incorporate enhanced proliferation resistance and energy security. (author)

  19. Culture medium pH influence on Gluconacetobacter physiology: Cellulose production rate and yield enhancement in presence of multiple carbon sources.

    Science.gov (United States)

    Yassine, Fatima; Bassil, Nathalie; Flouty, Roula; Chokr, Ali; Samrani, Antoine El; Boiteux, Gisèle; Tahchi, Mario El

    2016-08-01

    Gluconacetobacter genera are valued for bacterial cellulose (BC) and acetic acid production. BC is produced at optimal yields in classical microbiological media that are expensive for a large scale of production. In addition, BC usage for industrial purposes is limited due to low conversion rate into cellulose and to long incubation duration. In this paper, Gluconacetobacter isolated from apple vinegar was kinetically studied to evaluate cellulose production in presence of different carbon sources. Acetic and citric acid effect on Gluconacetobacter metabolism is clarified. It was shown that Gluconacetobacter uses glucose as a primary carbon source for cells growth and products formation. Acetic acid employment as a co-carbon source in Hestrin Schramm medium showed an increase of 17% in BC yield with a moderate decrease in the crystallite size of the resulting polymer. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. 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.

  1. Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells

    DEFF Research Database (Denmark)

    Rasmussen, Jeppe Grøndahl; Frøbert, Ole; Pilgaard, Linda

    2011-01-01

    Transplantation of mesenchymal stromal cells (MSC), including adipose tissue-derived stem cells (ASC), is a promising option in the treatment of vascular disease. Short-term hypoxic culture of MSC augments secretion of anti-apoptotic and angiogenic cytokines. We hypothesized that prolonged hypoxic...... (1% and 5% oxygen) culture and trypsinization would augment ASC expression of anti-apoptotic and angiogenic cytokines and increase the angiogenic potential of ASC-conditioned media....

  2. Cultural stereotypes as gatekeepers: increasing girls’ interest in computer science and engineering by diversifying stereotypes

    OpenAIRE

    Cheryan, Sapna; Master, Allison; Meltzoff, Andrew N.

    2015-01-01

    Despite having made significant inroads into many traditionally male-dominated fields (e.g., biology, chemistry), women continue to be underrepresented in computer science and engineering. We propose that students’ stereotypes about the culture of these fields—including the kind of people, the work involved, and the values of the field—steer girls away from choosing to enter them. Computer science and engineering are stereotyped in modern American culture as male-oriented fields that involve ...

  3. Medium-Chain Triglycerides in Combination with Leucine and Vitamin D Increase Muscle Strength and Function in Frail Elderly Adults in a Randomized Controlled Trial.

    Science.gov (United States)

    Abe, Sakiko; Ezaki, Osamu; Suzuki, Motohisa

    2016-05-01

    Sarcopenia, the loss of skeletal muscle mass, strength, and function, is common in elderly individuals but difficult to treat. A combination of nutrients was investigated to treat sarcopenia in very frail elderly adults. We enrolled 38 elderly nursing home residents (11 men and 27 women with a mean ± SD age of 86.6 ± 4.8 y) in a 3-mo randomized, controlled, single-blind, parallel group trial. The participants were randomly allocated to 3 groups. The first group received a daily l-leucine (1.2 g) and cholecalciferol (20 μg)-enriched supplement with 6 g medium-chain triglycerides (TGs) (MCTs) (LD + MCT); the second group received the same leucine and cholecalciferol-enriched supplement with 6 g long-chain TGs (LD + LCT); and the third group did not receive any supplements (control). The supplement and oils were taken at dinner, and changes in muscle mass, strength, and function were monitored. The increase in body weight in the LD + MCT (1.1 ± 1.0 kg) and LD + LCT (0.8 ± 1.1 kg) groups was greater than that in the control group (-0.5 ± 0.9 kg) (P elderly individuals. This trial was registered at the University Hospital Medical Information Network Clinical Trials Registry as UMIN000017567. © 2016 American Society for Nutrition.

  4. Reduced ultraviolet light transmission increases insecticide longevity in protected culture raspberry production.

    Science.gov (United States)

    Leach, Heather; Wise, John C; Isaacs, Rufus

    2017-12-01

    High tunnels are large protective structures used for season extension of many crops, including raspberries. These structures are often covered in plastic films to reduce and diffuse ultraviolet light transmission for pest and disease control, but this may also affect the photodegradation and efficacy of pesticides applied under these tunnels. We compared the residue levels of ten insecticides under three tunnel plastics with varying levels of UV transmission and open field conditions. Raspberry plants placed in research-scale tunnels were treated with insecticides and residues on fruit and foliage were monitored for one or two weeks in early 2015 and early and late 2016. Plastics that reduce UV transmission resulted in 50% greater residues of some insecticides compared to transparent plastics, and 60% compared to uncovered tunnels. This increased persistence of residues was evident within 1 day and remained consistently higher for up to 14 days. This pattern was demonstrated for multiple insecticides, including bifenthrin, esfenvalerate, imidacloprid, thiamethoxam, and spinosad. In contrast, the insecticide malathion degraded rapidly regardless of the plastic treatment, indicating less sensitivity to photodegradation. Bioassays using insecticide-treated leaves that were under UV-blocking plastic revealed higher mortality of the invasive fruit pest, Drosophila suzukii, compared to leaves that were uncovered. This indicates that the activity of pesticides under high tunnels covered in UV-reducing plastics may be prolonged, allowing for fewer insecticide applications and longer intervals between sprays. This information can be used to help optimize pest control in protected culture berry production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. 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.

  6. 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.

  7. 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.

  8. Co-Culture with Listeria monocytogenes within a Dual-Species Biofilm Community Strongly Increases Resistance of Pseudomonas putida to Benzalkonium Chloride

    Science.gov (United States)

    Giaouris, Efstathios; Chorianopoulos, Nikos; Doulgeraki, Agapi; Nychas, George-John

    2013-01-01

    Biofilm formation is a phenomenon occurring almost wherever microorganisms and surfaces exist in close proximity. This study aimed to evaluate the possible influence of bacterial interactions on the ability of Listeria monocytogenes and Pseudomonas putida to develop a dual-species biofilm community on stainless steel (SS), as well as on the subsequent resistance of their sessile cells to benzalkonium chloride (BC) used in inadequate (sub-lethal) concentration (50 ppm). The possible progressive adaptability of mixed-culture biofilms to BC was also investigated. To accomplish these, 3 strains per species were left to develop mixed-culture biofilms on SS coupons, incubated in daily renewable growth medium for a total period of 10 days, under either mono- or dual-species conditions. Each day, biofilm cells were exposed to disinfection treatment. Results revealed that the simultaneous presence of L. monocytogenes strongly increased the resistance of P. putida biofilm cells to BC, while culture conditions (mono-/dual-species) did not seem to significantly influence the resistance of L. monocytogenes biofilm cells. BC mainly killed L. monocytogenes cells when this was applied against the dual-species sessile community during the whole incubation period, despite the fact that from the 2nd day this community was mainly composed (>90%) of P. putida cells. No obvious adaptation to BC was observed in either L. monocytogenes or P. putida biofilm cells. Pulsed field gel electrophoresis (PFGE) analysis showed that the different strains behaved differently with regard to biofilm formation and antimicrobial resistance. Such knowledge on the physiological behavior of mixed-culture biofilms could provide the information necessary to control their formation. PMID:24130873

  9. Co-culture with Listeria monocytogenes within a dual-species biofilm community strongly increases resistance of Pseudomonas putida to benzalkonium chloride.

    Directory of Open Access Journals (Sweden)

    Efstathios Giaouris

    Full Text Available Biofilm formation is a phenomenon occurring almost wherever microorganisms and surfaces exist in close proximity. This study aimed to evaluate the possible influence of bacterial interactions on the ability of Listeria monocytogenes and Pseudomonas putida to develop a dual-species biofilm community on stainless steel (SS, as well as on the subsequent resistance of their sessile cells to benzalkonium chloride (BC used in inadequate (sub-lethal concentration (50 ppm. The possible progressive adaptability of mixed-culture biofilms to BC was also investigated. To accomplish these, 3 strains per species were left to develop mixed-culture biofilms on SS coupons, incubated in daily renewable growth medium for a total period of 10 days, under either mono- or dual-species conditions. Each day, biofilm cells were exposed to disinfection treatment. Results revealed that the simultaneous presence of L. monocytogenes strongly increased the resistance of P. putida biofilm cells to BC, while culture conditions (mono-/dual-species did not seem to significantly influence the resistance of L. monocytogenes biofilm cells. BC mainly killed L. monocytogenes cells when this was applied against the dual-species sessile community during the whole incubation period, despite the fact that from the 2nd day this community was mainly composed (>90% of P. putida cells. No obvious adaptation to BC was observed in either L. monocytogenes or P. putida biofilm cells. Pulsed field gel electrophoresis (PFGE analysis showed that the different strains behaved differently with regard to biofilm formation and antimicrobial resistance. Such knowledge on the physiological behavior of mixed-culture biofilms could provide the information necessary to control their formation.

  10. Cultural/Favorite Recipe Day: Strengthening Approaches to Increase Culturally Diverse Foods Served in Head Start Meals

    Science.gov (United States)

    Hoffman, Jessica A.; Agrawal, Tara; Carter, Sonia; Grinder, AnnMarie; Castaneda-Sceppa, Carmen

    2012-01-01

    One approach to halting the childhood obesity epidemic has been the modification of foods available to children during the school day. In recent years there has been an increased focus on obesity prevention efforts among children ages birth to 5 and the role of child care settings in prevention efforts. Head Start serves as an important venue for…

  11. Development of small and medium business (SMES) of mango dodol processing to increase the added value (a case study in Ujungjaya Village, Indramayu District, West Java)

    Science.gov (United States)

    Sulistyowati, L.; Pardian, P.; Syamsyiah, N.; Deliana, Y.

    2018-03-01

    In the national economic development in Indonesia, Small and Medium Enterprises (SMEs) become a priority to be developed, because SMEs can be the backbone of the populist economic system to reduce the problem of poverty. In addition, the development of SMEs is able to expand the economic base and can contribute to the increase of added value, in addition it would also serve to open employment opportunities in rural areas. Indramayu is one of the three mango production centers in West Java that face the problem that there are about 20% of the mangoes that is not worth selling. This opportunity is utilized by women who are members of KUB (Joint Business Group) to be processed into mango dodol at household scale. But this effort has not been widespread, only pioneered by a small portion of women. This study aims toobserve the driving force of women to participate in the processing of mango dodol, and whether the mango processing business to become mango dodol is profitable, also how much added value obtained. This study uses case study method with interview for data collection, participant observation and documentation study. While the data analysis technique using Hayami Value-added Method and descriptive analysis. The results revealed that the factors that affect the women’s participation in the processing of dodol is to increase family income, take advantage of spare time and take advantage of rejected mangoes. The added value obtained in mango dodol processing is Rp.50.600,00 per kilogram of input, with a value-added ratio of 52.8%. For the development of SMEs mangoes Training and socialization are needed for the good dodol processing and hygienic according to SOP (Standard Operational Procedure) from the relevant institutions, innovation in packaging, pioneering business partnerships with stores in the city of Indramayu and surrounding areas, and support financing from banks with an affordable interest rate.

  12. Transfer of human frozen-thawed embryos with further cleavage during culture increases pregnancy rates

    Directory of Open Access Journals (Sweden)

    Bharat V Joshi

    2010-01-01

    Full Text Available Aim: To compare the pregnancy rate following transfer of frozen-thawed embryos with or without overnight culture after thawing. Settings and Design: This is a retrospective analysis of frozen-thawed embryo transfer (FET cycles performed between January 2006 and December 2008. Materials and Methods: Out of 518 thaw cycles, 504 resulted in embryo transfers (ETs. Of the total FET cycles, 415 were performed after an overnight culture of embryos (group A; and in 89 cycles, ET was performed within 2 hours of embryo thawing (group B. Statistical Analysis: The data were statistically analyzed using chi-square test. Results: We observed that with FET, women ≤30 years of age had a significantly higher (P=0.003 pregnancy rate (PR=28.9% as compared to women >30 years of age (17.5%. A significantly higher (P<0.001FNx08 pregnancy rate was also observed in women receiving 3 frozen-thawed embryos (29% as compared to those who received less than 3 embryos (10.7%. The difference in PR between group A (PR=24.3% and group B (PR=20.3% was not statistically significant. However, within group A, ET with cleaved embryos showed significantly ( P≤0.01 higher pregnancy rate compared to the uncleaved embryos, depending on the number of cleaved embryos transferred. Conclusion: No significant difference was noticed between FETs made with transfer of embryos with overnight culture and those without culture. However, within the cultured group, transfer of embryos cleaved during overnight culture gave significantly higher PR than transfers without any cleavage.

  13. 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

  14. Increased susceptibility to in vitro ultraviolet B radiation in fibroblasts and lymphocytes cultured from systemic lupus erythematosus patients

    International Nuclear Information System (INIS)

    Golan, T.D.; Foltyn, V.; Roueff, A.

    1991-01-01

    Sunlight is known to induce exacerbations of systemic lupus erythematosus (SLE) but its mechanism remains unclear. We have previously reported that ultraviolet A (UVA) exposure induces an increase in total DNA synthesis (DS) in vitro but a decrease in unscheduled DNA repair synthesis (UDRS) of splenocytes of murine SLE strains. In order to investigate whether similar observations are characteristic of human SLE, peripheral blood lymphocytes (PBL) and dermal fibroblast (DF) cultures of 20 patients and 15 matched controls were exposed in vitro to UVA or UVB at different doses. Thirteen (65%) SLE DF cultures exposed to UVB light (12-24 J/m2) showed an increase in DS compared to paired unirradiated cultures. In contrast, UVB-irradiated DF from normal individuals had no significant increase in DS following UVB irradiation. When SLE DF were exposed to higher doses of UVB (48-96 J/m2), 90% of cultures showed a decrease in DS compared to only 20% in the control group. All of the SLE DF cultures showed a decrease of their unscheduled DNA repair capacity following UVB (24-48 J/m2) irradiation whereas no UDRS was apparent in 74% of controls under the same conditions. Similar findings regarding UDRS were observed in SLE PBL cultures and were also confirmed by autoradiography. UVA exposure (0-3840 J/m2) had no effect on DS nor on UDRS in DF or PBL cultured from SLE and controls. The relevance of these in vitro findings to the in vivo pathogenesis of the disease is discussed

  15. Culture-Based Contextual Learning to Increase Problem-Solving Ability of First Year University Student

    Science.gov (United States)

    Samo, Damianus Dao; Darhim; Kartasasmita, Bana G.

    2018-01-01

    The purpose of this study is to show the differences in problem-solving ability between first-year University students who received culture-based contextual learning and conventional learning. This research is a quantitative research using quasi-experimental research design. Samples were the First-year students of mathematics education department;…

  16. Measuring Collaboration and Communication to Increase Implementation of Evidence-Based Practices: The Cultural Exchange Inventory

    Science.gov (United States)

    Palinkas, Lawrence A.; Garcia, Antonio; Aarons, Gregory; Finno-Velasquez, Megan; Fuentes, Dahlia; Holloway, Ian; Chamberlain, Patricia

    2018-01-01

    The Cultural Exchange Inventory (CEI) is a 15-item instrument designed to measure the process (7 items) and outcomes (8 items) of exchanges of knowledge, attitudes and practices between members of different organisations collaborating in implementing evidence-based practice. We conducted principal axis factor analyses and parallel analyses of data…

  17. Exposing Students to Repeat Photography: Increasing Cultural Understanding on a Short-Term Study Abroad

    Science.gov (United States)

    Lemmons, Kelly K.; Brannstrom, Christian; Hurd, Danielle

    2014-01-01

    Traditionally, repeat photography has been used to analyze land cover change. This paper describes how repeat photography may be used as a tool to enhance the short-term study abroad experience by facilitating cultural interaction and understanding. We present evidence from two cases and suggest a five-step repeat photography method for educators…

  18. Digital Storytelling: A Method for Engaging Students and Increasing Cultural Competency

    Science.gov (United States)

    Grant, Natalie S.; Bolin, Brien L.

    2016-01-01

    Digital storytelling is explored as a method of engaging students in the development of media literacy and cultural competency. This paper describes the perceptions and experiences of 96 undergraduate students at a large Midwestern university, after completing a digital storytelling project in a semester-long diversity course. Digital storytelling…

  19. Effect of early addition of bone morphogenetic protein 5 (BMP5) to embryo culture medium on in vitro development and expression of developmentally important genes in bovine preimplantation embryos.

    Science.gov (United States)

    García, Elina V; Miceli, Dora C; Rizo, Gabriela; Valdecantos, Pablo A; Barrera, Antonio D

    2015-09-01

    blastocyst stage in the BMP5 group. Moreover, reverse transcription quantitative real-time polymerase chain reaction analysis showed a significant increase in the relative abundance of SOX2 in two-cell stage embryos, ID2 and OCT4 in eight-cell stage embryos, and NANOG and OCT4 in blastocysts derived from BMP5-treated embryos. In conclusion, our results report that early addition of BMP5 to the embryo culture medium had a positive effect on the blastocyst rate and affected the relative expression of BMP target and pluripotency genes, suggesting that BMP5 could play an important role in the preimplantation development of bovine embryos. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Eight-Shaped Hatching Increases the Risk of Inner Cell Mass Splitting in Extended Mouse Embryo Culture.

    Directory of Open Access Journals (Sweden)

    Zheng Yan

    Full Text Available Increased risk of monozygotic twinning (MZT has been shown to be associated with assisted reproduction techniques, particularly blastocyst culture. Interestingly, inner cell mass (ICM splitting in human '8'-shaped hatching blastocysts that resulted in MZT was reported. However, the underlying cause of MZT is not known. In this study, we investigated in a mouse model whether in vitro culture leads to ICM splitting and its association with hatching types. Blastocyst hatching was observed in: (i in vivo developed blastocysts and (ii-iii in vitro cultured blastocysts following in vivo or in vitro fertilization. We found that '8'-shaped hatching occurred with significantly higher frequency in the two groups of in vitro cultured blastocysts than in the group of in vivo developed blastocysts (24.4% and 20.4% versus 0.8%, respectively; n = 805, P < 0.01. Moreover, Oct4 immunofluorescence staining was performed to identify the ICM in the hatching and hatched blastocysts. Scattered and split distribution of ICM cells was observed around the small zona opening of '8'-shaped hatching blastocysts. This occurred at a high frequency in the in vitro cultured groups. Furthermore, we found more double OCT4-positive masses, suggestive of increased ICM splitting in '8'-shaped hatching and hatched blastocysts than in 'U'-shaped hatching and hatched blastocysts (12.5% versus 1.9%, respectively; n = 838, P < 0.01. Therefore, our results demonstrate that extended in vitro culture can cause high frequencies of '8'-shaped hatching, and '8'-shaped hatching that may disturb ICM herniation leading to increased risk of ICM splitting in mouse blastocysts. These results may provide insights into the increased risk of human MZT after in vitro fertilization and blastocyst transfer.

  1. A Conceptual Framework for Inclusive Digital Storytelling to Increase Diversity and Motivation for Cultural Tourism in Thailand.

    Science.gov (United States)

    Kasemsarn, Kittichai; Nickpour, Farnaz

    2016-01-01

    Cultural tourism is considered to be a niche market and little attention has been paid to it, as compared with mass tourism. Moreover, visitors have little motivation to visit actual historical sites and read the story displayed behind the exhibitions. These issues highlight a good opportunity to increase further potential extended tourism and increase the motivation of visitors. To broaden and increase the potential market, this study applies inclusive design principles as 'understanding and designing for diversity' and presents reports on the first study. To increase the motivation of tourists, this study adopts digital storytelling as 'the guideline to increase motivation' and illustrates this in the second study.

  2. Whey and Casein Proteins and Medium-Chain Saturated Fatty Acids from Milk Do Not Increase Low-Grade Inflammation in Abdominally Obese Adults.

    Science.gov (United States)

    Bohl, Mette; Bjørnshave, Ann; Gregersen, Søren; Hermansen, Kjeld

    2016-01-01

    Low-grade inflammation is involved in the development of diabetes and cardiovascular disease (CVD). Inflammation can be modulated by dietary factors. Dairy products are rich in saturated fatty acids (SFA), which are known to possess pro-inflammatory properties. However, different fatty acid compositions may exert different effects. Other components such as milk proteins may exert anti-inflammatory properties which may compensate for the potential negative effects of SFAs. Generally, the available data suggest a neutral role of dairy product consumption on inflammation. To investigate the effects of, and potential interaction between, a dietary supplementation with whey protein and milk fat, naturally enriched in medium-chain SFA (MC-SFA), on inflammatory markers in abdominal obese adults. The study was a 12-week, randomized, double-blinded, intervention study. Sixty-three adults were equally allocated to one of four groups which received a supplement of either 60 g/day whey or 60 g/day casein plus 63 g/day milk fat either high or low in MC-SFA content. Fifty-two subjects completed the study. Before and after the intervention, changes in plasma interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1RA), high-sensitive C-reactive protein (hsCRP), adiponectin, and monocyte chemoattractant protein-1 (MCP-1) were measured. Changes in inflammatory genes in the subcutaneous adipose tissue were also documented. There were no differences in circulating inflammatory markers between protein types or fatty acid compositions in abdominally obese subjects, with the exception of an increase in adiponectin in response to high compared to low MC-SFA consumption in women. We found that combined dairy proteins and MC-SFAs influenced inflammatory gene expression in adipose tissue, while no effect was detected by dairy proteins or MC-SFA per se. Whey protein compared with casein and MC-SFA-enriched milk fat did not alter circulating markers of low-grade inflammation in

  3. Does high-stakes testing increase cultural capital among low-income and racial minority students?

    Directory of Open Access Journals (Sweden)

    Won-Pyo Hong

    2008-03-01

    Full Text Available This article draws on research from Texas and Chicago to examine whether high-stakes testing enables low-income and racial minority students to acquire cultural capital. While students' performance on state or district tests rose after the implementation of high-stakes testing and accountability policies in Texas and Chicago in the 1990s, several studies indicate that these policies seemed to have had deleterious effects on curriculum, instruction, the percentage of students excluded from the tests, and student dropout rates. As a result, the policies seemed to have had mixed effects on students' opportunities to acquire embodied and institutionalized cultural capital. These findings are consistent with the work of Shepard (2000, Darling-Hammond (2004a, and others who have written of the likely negative repercussions of high-stakes testing and accountability policies.

  4. Chronic Treatment with a Water-Soluble Extract from the Culture Medium of Ganoderma lucidum Mycelia Prevents Apoptosis and Necroptosis in Hypoxia/Ischemia-Induced Injury of Type 2 Diabetic Mouse Brain

    Directory of Open Access Journals (Sweden)

    Meiyan Xuan

    2015-01-01

    Full Text Available Type 2 diabetes mellitus has been known to increase systemic oxidative stress by chronic hyperglycemia and visceral obesity and aggravate cerebral ischemic injury. On the basis of our previous study regarding a water-soluble extract from the culture medium of Ganoderma lucidum mycelia (designed as MAK, which exerts antioxidative and neuroprotective effects, the present study was conducted to evaluate the preventive effects of MAK on apoptosis and necroptosis (a programmed necrosis induced by hypoxia/ischemia (H/I in type 2 diabetic KKAy mice. H/I was induced by a combination of unilateral common carotid artery ligation with hypoxia (8% O2 for 20 min and subsequent reoxygenation. Pretreatment with MAK (1 g/kg, p.o. for a week significantly reduced H/I-induced neurological deficits and brain infarction volume assessed at 24 h of reoxygenation. Histochemical analysis showed that MAK significantly suppressed superoxide production, neuronal cell death, and vacuolation in the ischemic penumbra, which was accompanied by a decrease in the numbers of TUNEL- or cleaved caspase-3-positive cells. Furthermore, MAK decreased the expression of receptor-interacting protein kinase 3 mRNA and protein, a key molecule for necroptosis. These results suggest that MAK confers resistance to apoptotic and necroptotic cell death and relieves H/I-induced cerebral ischemic injury in type 2 diabetic mice.

  5. Carbon Tetrachloride Increases Intracellular Calcium in Rat Liver and Hepatocyte Cultures

    Science.gov (United States)

    1986-05-12

    tible to destruction by CC14 ( Head ~ al., 1981). Thus, the activation of CC14 to a toxic moiety clearly depends upon metabolism by one or more... embryologic development (Wyllie, 1986). In contrast, Toyo-oka et al. (1985) could not establish that phospholipase& or proteases were involved in ischemic...D. M. Bissell, and u. A Meyer. (1977) Drug Metabolism in Adult Rat Hepatocyte& in Primary Monolayer Culture. Gastroenterology 72:1232-1239. Head , B

  6. Mechanisms of Change in the ARC Organizational Strategy: Increasing Mental Health Clinicians' EBP Adoption Through Improved Organizational Culture and Capacity.

    Science.gov (United States)

    Williams, Nathaniel J; Glisson, Charles; Hemmelgarn, Anthony; Green, Philip

    2017-03-01

    The development of efficient and scalable implementation strategies in mental health is restricted by poor understanding of the change mechanisms that increase clinicians' evidence-based practice (EBP) adoption. This study tests the cross-level change mechanisms that link an empirically-supported organizational strategy for supporting implementation (labeled ARC for Availability, Responsiveness, and Continuity) to mental health clinicians' EBP adoption and use. Four hundred seventy-five mental health clinicians in 14 children's mental health agencies were randomly assigned to the ARC intervention or a control condition. Measures of organizational culture, clinicians' intentions to adopt EBPs, and job-related EBP barriers were collected before, during, and upon completion of the three-year ARC intervention. EBP adoption and use were assessed at 12-month follow-up. Multilevel mediation analyses tested changes in organizational culture, clinicians' intentions to adopt EBPs, and job-related EBP barriers as linking mechanisms explaining the effects of ARC on clinicians' EBP adoption and use. ARC increased clinicians' EBP adoption (OR = 3.19, p = .003) and use (81 vs. 56 %, d = .79, p = .003) at 12-month follow-up. These effects were mediated by improvement in organizational proficiency culture leading to increased clinician intentions to adopt EBPs and by reduced job-related EBP barriers. A combined mediation analysis indicated the organizational culture-EBP intentions mechanism was the primary carrier of ARC's effects on clinicians' EBP adoption and use. ARC increases clinicians' EBP adoption and use by creating proficient organizational cultures that increase clinicians' intentions to adopt EBPs.

  7. Mechanisms of change in the ARC organizational strategy: Increasing mental health clinicians’ EBP adoption through improved organizational culture and capacity

    Science.gov (United States)

    Williams, Nathaniel J.; Glisson, Charles; Hemmelgarn, Anthony; Green, Philip

    2016-01-01

    Background The development of efficient and scalable implementation strategies in mental health is restricted by poor understanding of the change mechanisms that increase clinicians’ evidence-based practice (EBP) adoption. This study tests the cross-level change mechanisms that link an empirically-supported organizational strategy for supporting implementation (labeled ARC for Availability, Responsiveness, and Continuity) to mental health clinicians’ EBP adoption and use. Method Four hundred seventy five mental health clinicians in 14 children’s mental health agencies were randomly assigned to the ARC intervention or a control condition. Measures of organizational culture, clinicians’ intentions to adopt EBPs, and job-related EBP barriers were collected before, during, and upon completion of the three-year ARC intervention. EBP adoption and use were assessed at 12-month follow-up. Multilevel mediation analyses tested changes in organizational culture, clinicians’ intentions to adopt EBPs, and job-related EBP barriers as linking mechanisms explaining the effects of ARC on clinicians’ EBP adoption and use. Results ARC increased clinicians’ EBP adoption (OR = 3.19, p = .003) and use (81% vs. 56%, d = .79, p = .003) at 12-month follow-up. These effects were mediated by improvement in organizational proficiency culture leading to increased clinician intentions to adopt EBPs and by reduced job-related EBP barriers. A combined mediation analysis indicated the organizational culture-EBP intentions mechanism was the primary carrier of ARC’s effects on clinicians’ EBP adoption and use. Conclusions ARC increases clinicians’ EBP adoption and use by creating proficient organizational cultures that increase clinicians’ intentions to adopt EBPs. PMID:27236457

  8. Macroautophagy-generated increase of lysosomal amyloid β-protein mediates oxidant-induced apoptosis of cultured neuroblastoma cells

    DEFF Research Database (Denmark)

    Zheng, Lin; Terman, Alexei; Hallbeck, Martin

    2011-01-01

    and accumulation of Aβ within lysosomes, induced apoptosis in differentiated SH-SY5Y neuroblastoma cells. Cells under hyperoxia showed: (1) increased numbers of autophagic vacuoles that contained amyloid precursor protein (APP) as well as Aβ monomers and oligomers, (2) increased reactive oxygen species production...... and resulting lysosomal Aβ accumulation are essential for oxidant-induced apoptosis in cultured neuroblastoma cells and provide additional support for the interactive role of oxidative stress and the lysosomal system in AD-related neurodegeneration....

  9. Early increase and late decrease of purkinje cell dendritic spine density in prion-infected organotypic mouse cerebellar cultures.

    Science.gov (United States)

    Campeau, Jody L; Wu, Gengshu; Bell, John R; Rasmussen, Jay; Sim, Valerie L

    2013-01-01

    Prion diseases are infectious neurodegenerative diseases associated with the accumulation of protease-resistant prion protein, neuronal loss, spongiform change and astrogliosis. In the mouse model, the loss of dendritic spines is one of the earliest pathological changes observed in vivo, occurring 4-5 weeks after the first detection of protease-resistant prion protein in the brain. While there are cell culture models of prion infection, most do not recapitulate the neuropathology seen in vivo. Only the recently developed prion organotypic slice culture assay has been reported to undergo neuronal loss and the development of some aspects of prion pathology, namely small vacuolar degeneration and tubulovesicular bodies. Given the rapid replication of prions in this system, with protease-resistant prion protein detectable by 21 days, we investigated whether the dendritic spine loss and altered dendritic morphology seen in prion disease might also develop within the lifetime of this culture system. Indeed, six weeks after first detection of protease-resistant prion protein in tga20 mouse cerebellar slice cultures infected with RML prion strain, we found a statistically significant loss of Purkinje cell dendritic spines and altered dendritic morphology in infected cultures, analogous to that seen in vivo. In addition, we found a transient but statistically significant increase in Purkinje cell dendritic spine density during infection, at the time when protease-resistant prion protein was first detectable in culture. Our findings support the use of this slice culture system as one which recapitulates prion disease pathology and one which may facilitate study of the earliest stages of prion disease pathogenesis.

  10. An innovative program to increase safety culture for workers on a nuclear power plant

    International Nuclear Information System (INIS)

    Schryvers, Vincent

    2007-01-01

    Full text: To implement the WENRA harmonized guidelines and the IAEA reference guides, Electrabel has recently introduced a major training program for both its own staff and the contractors working on the sites of its Nuclear Power Plants. This training program stresses the importance of safety culture on both theoretical and practical level and is mostly focused on the behavioural aspects during activities performed at the site of a Nuclear Power Plant. Further emphasis is put on radiation protection, industrial safety, environmental protection and explosion prevention. The training scheme for both the staff of Electrabel and contractors typically contains a theoretical part introducing the basic concepts of nuclear safety and safety culture and a practical exercise in a simulated environment. A novel element in the training cycle is the use of a simulated environment, where the actual working conditions in the nuclear part of the installation are simulated. This mock-up installation enables the workers to train the nuclear safety constraints linked to the actual installation and to enhance safety culture by responding on simulated problems and changing conditions possibly being encountered during an intervention at the real working site. To analyze the behaviour of the future workers, the activities are videotaped and commented for further improvement. A refresh of the training courses is implemented after 3 years.Although this training program has only been in operation for just 6 months, the response of the contractors and the staff to this training has been enthusiastic. At this moment, more than 1.000 workers have successfully completed the training course. (author)

  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. Concentrated fed-batch cell culture increases manufacturing capacity without additional volumetric capacity.

    Science.gov (United States)

    Yang, William C; Minkler, Daniel F; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

    2016-01-10

    Biomanufacturing factories of the future are transitioning from large, single-product facilities toward smaller, multi-product, flexible facilities. Flexible capacity allows companies to adapt to ever-changing pipeline and market demands. Concentrated fed-batch (CFB) cell culture enables flexible manufacturing capacity with limited volumetric capacity; it intensifies cell culture titers such that the output of a smaller facility can rival that of a larger facility. We tested this hypothesis at bench scale by developing a feeding strategy for CFB and applying it to two cell lines. CFB improved cell line A output by 105% and cell line B output by 70% compared to traditional fed-batch (TFB) processes. CFB did not greatly change cell line A product quality, but it improved cell line B charge heterogeneity, suggesting that CFB has both process and product quality benefits. We projected CFB output gains in the context of a 2000-L small-scale facility, but the output was lower than that of a 15,000-L large-scale TFB facility. CFB's high cell mass also complicated operations, eroded volumetric productivity, and showed our current processes require significant improvements in specific productivity in order to realize their full potential and savings in manufacturing. Thus, improving specific productivity can resolve CFB's cost, scale-up, and operability challenges. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Development of technology for plantlet propagation by tissue culture. (3). Strawberry callus growth and changes in pH in liquid medium; Soshiki baiyo ni yoru shubyo tairyo zoshoku gijutsu no kaihatsu. (3). Ekitai baiyo ni okeru ichigo callus no zoshoku to baichi no pH henka

    Energy Technology Data Exchange (ETDEWEB)

    Yoshihara, T; Hanyo, H [Central Research Inst. of Electric Power Industry, Tokyo (Japan)

    1991-02-01

    Development of strawberry cultivation plants is described as a part of night-time power utilization activities for power load levelling. Strawberry calluses (undifferent tissue mass) cultured in a liquid medium reach the fastest growth period in one week and the steady state in four weeks. The callus growth shows the maximum value at this time, which was 20 times as much of the seedling. The medium pH changed in a range from 4 to 7. If the initial pH is 4.0 or higher, no difference is created in the callus propagation in the steady state period, but at 3.0, no propagation whatsoever. The pH after the fastest growth period converged to a range from 6.0 to 7.0, with the exception of initial pH at 3.0. The medium pH decreased as a result of pre-culture heating sterilization, formation of iron phosphate due to light irradiation, and organic acid release during the initial growth phase. The pH increased because of difference in the speed of absorbing ammonium and nitric acid during the later growth phase. The growth efficiency of 20 times is about the same as other plants. Since the pH change is maintained within the range from 4 to 7, which causes no difference in in growth, there is no need of adjusting the pH within this range. 18 refs., 15 figs., 3 tabs.

  14. USING ONLINE MARKETING TO INCREASE PARTICIPATION IN A WEB-BASED CONTINUING MEDICAL EDUCATION CULTURAL COMPETENCE CURRICULUM

    Science.gov (United States)

    Estrada, Carlos A.; Krishnamoorthy, Periyakaruppan; Smith, Ann; Staton, Lisa; Korf, Michele J.; Allison, Jeroan J.; Houston, Thomas K.

    2012-01-01

    Introduction CME providers may be interested in identifying effective marketing strategies to direct users to specific content. The use of online advertisements to recruit participants for clinical trials, public health programs, and Continuing Medical Education (CME) has been shown to be effective in some but not all studies. The purpose of this study was to compare the impact of two marketing strategies in the context of an online CME cultural competence curriculum (www.c-comp.org). Methods In an interrupted time-series quasi-experimental design, two marketing strategies were tested: a) wide dissemination to relevant organizations over a period of approximately four months, and b) Internet paid search using Google Ads (five consecutive eight-week periods--control 1, cultural/ CME advertisement, control 2, hypertension/ content advertisement, control 3). Outcome measures were CME credit requests, Web traffic (visits per day, page views, pages viewed per visit), and cost. Results Overall, the site was visited 19,156 times and 78,160 pages were viewed. During the wide dissemination phase, the proportion of visits requesting CME credit decreased between the first (5.3%) and second halves (3.3%) of this phase (p= .04). During the Internet paid search phase, the proportion of visits requesting CME credit was highest during the cultural/ CME advertisement period (control 1, 1.4%; cultural/CME ad, 4.3%; control 2, 1.5%; hypertension/content ad, 0.6%; control 3, 0.8%; p<.001). All measures of Web traffic changed during the Internet paid search phase (p<.01); however, changes were independent of the advertisement periods. The incremental cost for the cultural advertisement per CME credit requested was $0.64US. Discussion Internet advertisement focusing on cultural competence and CME was associated with about a three-fold increase in requests for CME credit at an incremental cost of under $1; however, Web traffic changes were independent of the advertisement strategy. PMID

  15. Resveratrol increases nucleus pulposus matrix synthesis through activating the PI3K/Akt signaling pathway under mechanical compression in a disc organ culture.

    Science.gov (United States)

    Han, Xiaorui; Leng, Xiaoming; Zhao, Man; Wu, Mei; Chen, Amei; Hong, Guoju; Sun, Ping

    2017-12-22

    Disc nucleus pulposus (NP) matrix homeostasis is important for normal disc function. Mechanical overloading seriously decreases matrix synthesis and increases matrix degradation. The present study aims to investigate the effects of resveratrol on disc NP matrix homeostasis under a relatively high-magnitude mechanical compression and the potential mechanism underlying this process. Porcine discs were perfusion-cultured and subjected to a relatively high-magnitude mechanical compression (1.3 MPa at a frequency of 1.0 Hz for 2 h once per day) for 7 days in a mechanically active bioreactor. The non-compressed discs were used as controls. Resveratrol was added along with culture medium to observe the effects of resveratrol on NP matrix synthesis under mechanical load respectively. NP matrix synthesis was evaluated by histology, biochemical content (glycosaminoglycan (GAG) and hydroxyproline (HYP)), and expression of matrix macromolecules (aggrecan and collagen II). Results showed that this high-magnitude mechanical compression significantly decreased NP matrix content, indicated by the decreased staining intensity of Alcian Blue and biochemical content (GAG and HYP), and the down-regulated expression of NP matrix macromolecules (aggrecan and collagen II). Further analysis indicated that resveratrol partly stimulated NP matrix synthesis and increased activity of the PI3K/Akt pathway in a dose-dependent manner under mechanical compression. Together, resveratrol is beneficial for disc NP matrix synthesis under mechanical overloading, and the activation of the PI3K/Akt pathway may participate in this regulatory process. Resveratrol may be promising to regenerate mechanical overloading-induced disc degeneration. © 2017 The Author(s).

  16. The increase in radioresistance of Chinese hamster cells cultured as spheroids is correlated to changes in nuclear morphology

    International Nuclear Information System (INIS)

    Gordon, D.J.; Milner, A.E.; Beaney, R.P.; Grdina, D.J.; Vaughan, A.T.

    1990-01-01

    Chinese hamster V79 cells grown as spheroids in roller culture are more radioresistant than those grown as monolayers. The supercoiled structure of chromatin, as salt-extracted nucleoids, has been examined using flow cytometry. Irradiated viable cells from spheroid culture contain restraints to supercoil relaxation that are absent in monolayer cells. Further analysis of the chromatin organization from each growth form shows that the radioresistant spheroid cells contain a DNA-protein matrix that is more resistant to detergent-induced degradation. The increase in structural integrity may be due to the retention of a 55-60 kDa protein that is apparent in the nucleoids of spheroid, but not monolayer cells. The increase in structural integrity of the spheroid cells may explain their greater radioresistance by providing a more stable platform for high-fidelity DNA damage repair

  17. Increased adipogenesis in cultured embryonic chondrocytes and in adult bone marrow of dominant negative Erg transgenic mice.

    Directory of Open Access Journals (Sweden)

    Sébastien Flajollet

    Full Text Available In monolayer culture, primary articular chondrocytes have an intrinsic tendency to lose their phenotype during expansion. The molecular events underlying this chondrocyte dedifferentiation are still largely unknown. Several transcription factors are important for chondrocyte differentiation. The Ets transcription factor family may be involved in skeletal development. One family member, the Erg gene, is mainly expressed during cartilage formation. To further investigate the potential role of Erg in the maintenance of the chondrocyte phenotype, we isolated and cultured chondrocytes from the rib cartilage of embryos of transgenic mice that express a dominant negative form of Erg (DN-Erg during cartilage formation. DN-Erg expression in chondrocytes cultured for up to 20 days did not affect the early dedifferentiation usually observed in cultured chondrocytes. However, lipid droplets accumulated in DN-Erg chondrocytes, suggesting adipocyte emergence. Transcriptomic analysis using a DNA microarray, validated by quantitative RT-PCR, revealed strong differential gene expression, with a decrease in chondrogenesis-related markers and an increase in adipogenesis-related gene expression in cultured DN-Erg chondrocytes. These results indicate that Erg is involved in either maintaining the chondrogenic phenotype in vitro or in cell fate orientation. Along with the in vitro studies, we compared adipocyte presence in wild-type and transgenic mice skeletons. Histological investigations revealed an increase in the number of adipocytes in the bone marrow of adult DN-Erg mice even though no adipocytes were detected in embryonic cartilage or bone. These findings suggest that the Ets transcription factor family may contribute to the homeostatic balance in skeleton cell plasticity.

  18. 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...

  19. Role of Changes in Cell Fatty Acids Composition in the Increasing of Frost Resistance of Winter Wheat Suspension Culture

    Directory of Open Access Journals (Sweden)

    I.V. Lyubushkina

    2013-11-01

    Full Text Available Influences of low temperatures (4 and 8 ° С on the frost tolerance and fatty acid compositions of cells in a winter wheat suspension culture have been studied. It has been found that treatment of the culture with 4 °C (7 days did not protect cells from subsequent freezing temperature action (-8 °С, 6 h and was not accompanied significant changes in the fatty acid composition. On the contrary, the treatment of the culture with the temperature 8 °C (7 days prevented the death caused by freezing temperature and the content of saturated fatty acids decreased: pentadecanoic acid (by 35,0%, palmitic acid (by 19,9% and stearic acid (by 65,4%, and the content of α-linolenic acid increased by 94%. That was the cause of the double bond index (DBI increase by 16%. The role of fatty acids composition changes in the process of increasing frost tolerance in plants are discussed.

  20. 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...

  1. Increasing clinicians' EBT exploration and preparation behavior in youth mental health services by changing organizational culture with ARC.

    Science.gov (United States)

    Glisson, Charles; Williams, Nathaniel J; Hemmelgarn, Anthony; Proctor, Enola; Green, Philip

    2016-01-01

    Clinician EBT exploration and preparation behavior is essential to the ongoing implementation of new EBTs. This study examined the effect of the ARC organizational intervention on clinician EBT exploration and preparation behavior and assessed the mediating role of organizational culture as a linking mechanism. Fourteen community mental health agencies that serve youth in a major Midwestern metropolis along with 475 clinicians who worked in those agencies, were randomly assigned to either the three-year ARC intervention or control condition. Organizational culture was assessed with the Organizational Social Context (OSC) measure at baseline and follow-up. EBT exploration and preparation behavior was measured as clinician participation in nine separate community EBT workshops held over a three-year period. There was 69 percent greater odds (OR = 1.69, p organizational culture mediated the positive effect of the ARC intervention on clinicians' workshop attendance (a × b = .21; 95% CI:LL = .05, UL = .40). Organizational interventions that create proficient mental health agency cultures can increase clinician EBT exploration and preparation behavior that is essential to the ongoing implementation of new EBTs in community youth mental health settings. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Changing the Educational Culture of the Home to Increase Student Success at School

    Directory of Open Access Journals (Sweden)

    Kenneth Leithwood

    2015-09-01

    Full Text Available Parent involvement in their children’s learning is widely acknowledged as having a positive effect on student academic success. Of particular relevance is the finding that the influence of parent engagement can mitigate differences in socioeconomic status (SES and family background. Family background is a multi-dimensional concept that includes the family’s “educational culture” (including for example, parenting style, parental expectations for children’s work at school, direct instructional support for school learning, active parent interest in the school’s curriculum, and the monitoring of children’s engagement with their school work. It is these features of a child’s home environment that directly influence much of the social and intellectual capital students need to be successful at school. This paper summarizes a quasi-experimental field study which explored the relative effects of alternative types of school interventions on parent engagement. All of these interventions aimed to further engage parents in the education of their children as a means of both improving student achievement and closing gaps in achievement for students living primarily in challenging social and economic circumstances. Initiatives by school staffs aimed at helping those families struggling to build productive educational cultures in their homes would appear to be a very promising strategy for closing achievement gaps between advantaged and disadvantaged students. The study provides eight lessons other districts might take heed of as they embark on their own parent engagement interventions.

  3. Changing the system by changing the workforce: employing consumers to increase access, cultural diversity, and engagement.

    Science.gov (United States)

    Wenz-Gross, Melodie; Irsfeld, Toni DuBrino; Twomey, Tammy; Perez, Ana; Thompson, Judith; Wally, Martha; Colleton, Barbara; Kroell, Christine; McKeown, Steven K; Metz, Peter

    2012-06-01

    Services to families have traditionally been delivered in a medical model. This presents challenges including workforce shortages, lack of cultural diversity, lack of training in strength-based work, and difficulty in successfully engaging and retaining families in the therapy process. The system of care (SOC) effort has worked to establish formal roles for caregivers in SOC to improve services. This paper provides an example of one community's efforts to change the SOC by expanding the roles available to caregivers in creating systems change. It describes the model developed by Communities of Care (CoC), a SOC in Central Massachusetts, and its evolution over a 10 year period. First person accounts by system partners, caregivers hired into professional roles as well as a family receiving services, demonstrate how hiring caregivers at all levels can change systems and change lives, not only for those being served but for the caregiver/professionals doing the work. It also demonstrates, however, that change at the system level is incremental, takes time, and can be fleeting unless an ongoing effort is made to support and sustain those changes.

  4. 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

  5. Fibroblast-Derived Extracellular Matrices: An Alternative Cell Culture System That Increases Metastatic Cellular Properties.

    Directory of Open Access Journals (Sweden)

    Michael T Scherzer

    Full Text Available Poor survival rates from lung cancer can largely be attributed to metastatic cells that invade and spread throughout the body. The tumor microenvironment (TME is composed of multiple cell types, as well as non-cellular components. The TME plays a critical role in the development of metastatic cancers by providing migratory cues and changing the properties of the tumor cells. The Extracellular Matrix (ECM, a main component of the TME, has been shown to change composition during tumor progression, contributing to cancer cell invasion and survival away from the primary cancer site. Although the ECM is well-known to influence the fate of tumor progression, little is known about the molecular mechanisms that are affected by the cancer cell-ECM interactions. It is imperative that these mechanisms are elucidated in order to properly understand and prevent lung cancer dissemination. However, common in vitro studies do not incorporate these interactions into everyday cell culture assays. We have adopted a model that examines decellularized human fibroblast-derived ECM as a 3-dimensional substrate for growth of lung adenocarcinoma cell lines. Here, we have characterized the effect of fibroblast-derived matrices on the properties of various lung-derived epithelial cell lines, including cancerous and non-transformed cells. This work highlights the significance of the cell-ECM interaction and its requirement for incorporation into in vitro experiments. Implementation of a fibroblast-derived ECM as an in vitro technique will provide researchers with an important factor to manipulate to better recreate and study the TME.

  6. Variant at serotonin transporter gene predicts increased imitation in toddlers: relevance to the human capacity for cumulative culture.

    Science.gov (United States)

    Schroeder, Kari Britt; Asherson, Philip; Blake, Peter R; Fenstermacher, Susan K; Saudino, Kimberly J

    2016-04-01

    Cumulative culture ostensibly arises from a set of sociocognitive processes which includes high-fidelity production imitation, prosociality and group identification. The latter processes are facilitated by unconscious imitation or social mimicry. The proximate mechanisms of individual variation in imitation may thus shed light on the evolutionary history of the human capacity for cumulative culture. In humans, a genetic component to variation in the propensity for imitation is likely. A functional length polymorphism in the serotonin transporter gene, the short allele at 5HTTLPR, is associated with heightened responsiveness to the social environment as well as anatomical and activational differences in the brain's imitation circuity. Here, we evaluate whether this polymorphism contributes to variation in production imitation and social mimicry. Toddlers with the short allele at 5HTTLPR exhibit increased social mimicry and increased fidelity of demonstrated novel object manipulations. Thus, the short allele is associated with two forms of imitation that may underlie the human capacity for cumulative culture. The short allele spread relatively recently, possibly due to selection, and its frequency varies dramatically on a global scale. Diverse observations can be unified via conceptualization of 5HTTLPR as influencing the propensity to experience others' emotions, actions and sensations, potentially through the mirror mechanism. © 2016 The Author(s).

  7. 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.

  8. Increased FasL expression correlates with apoptotic changes in granulocytes cultured with oxidized clozapine

    International Nuclear Information System (INIS)

    Husain, Zaheed; Almeciga, Ingrid; Delgado, Julio C.; Clavijo, Olga P.; Castro, Januario E.; Belalcazar, Viviana; Pinto, Clara; Zuniga, Joaquin; Romero, Viviana; Yunis, Edmond J.

    2006-01-01

    Clozapine has been associated with a 1% incidence of agranulocytosis. The formation of an oxidized intermediate clozapine metabolite has been implicated in direct polymorphonuclear (PMN) toxicity. We utilized two separate systems to analyze the role of oxidized clozapine in inducing apoptosis in treated cells. Human PMN cells incubated with clozapine (0-10 μM) in the presence of 0.1 mM H 2 O 2 demonstrated a progressive decrease of surface CD16 expression along with increased apoptosis. RT-PCR analysis showed decreased CD16 but increased FasL gene expression in clozapine-treated PMN cells. No change in constitutive Fas expression was observed in treated cells. In HL-60 cells induced to differentiate with retinoic acid (RA), a similar increase in FasL expression, but no associated changes in CD16 gene expression, was observed following clozapine treatments. Our results demonstrate increased FasL gene expression in oxidized clozapine-induced apoptotic neutrophils suggesting that apoptosis in granulocytes treated with clozapine involves Fas/FasL interaction that initiates a cascade of events leading to clozapine-induced agranulocytosis

  9. 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

  10. Ablation of the mitochondrial complex IV assembly protein Surf1 leads to increased expression of the UPRMT and increased resistance to oxidative stress in primary cultures of fibroblasts

    Directory of Open Access Journals (Sweden)

    Gavin Pharaoh

    2016-08-01

    Full Text Available Mice deficient in the electron transport chain (ETC complex IV assembly protein SURF1 have reduced assembly and activity of cytochrome c oxidase that is associated with an upregulation of components of the mitochondrial unfolded protein response (UPRMT and increased mitochondrial number. We hypothesized that the upregulation of proteins associated with the UPRMT in response to reduced cytochrome c oxidase activity in Surf1−/− mice might contribute to increased stress resistance. To test this hypothesis we asked whether primary cultures of fibroblasts from Surf1−/− mice exhibit enhanced resistance to stressors compared to wild-type fibroblasts. Here we show that primary dermal fibroblasts isolated from Surf1−/− mice have increased expression of UPRMT components ClpP and Hsp60, and increased expression of Lon protease. Fibroblasts from Surf1−/− mice are significantly more resistant to cell death caused by oxidative stress induced by paraquat or tert-Butyl hydroperoxide compared to cells from wild-type mice. In contrast, Surf1−/− fibroblasts show no difference in sensitivity to hydrogen peroxide stress. The enhanced cell survival in response to paraquat or tert-Butyl hydroperoxide in Surf1−/− fibroblasts compared to wild-type fibroblasts is associated with induced expression of Lon, ClpP, and Hsp60, increased maximal respiration, and increased reserve capacity as measured using the Seahorse Extracellular Flux Analyzer. Overall these data support a protective role for the activation of the UPRMT in cell survival.

  11. Increased production of megakaryocytes near purity from cord blood CD34+ cells using a short two-phase culture system.

    Science.gov (United States)

    Boyer, Lucie; Robert, Amélie; Proulx, Chantal; Pineault, Nicolas

    2008-03-20

    Expansion of hematopoietic progenitor cells (HPC) ex vivo remains an important focus in fundamental and clinical research. The aim of this study was to determine whether the implementation of such expansion phase in a two-phase culture strategy prior to the induction of megakaryocyte (Mk) differentiation would increase the yield of Mks produced in cultures. Toward this end, we first characterized the functional properties of five cytokine cocktails to be tested in the expansion phase on the growth and differentiation kinetics of CD34+-enriched cells, and on their capacity to expand clonogenic progenitors in cultures. Three of these cocktails were chosen based on their reported ability to induce HPC expansion ex vivo, while the other two represented new cytokine combinations. These analyses revealed that none of the cocktails tested could prevent the differentiation of CD34+ cells and the rapid expansion of lineage-positive cells. Hence, we sought to determine the optimum length of time for the expansion phase that would lead to the best final Mk yields. Despite greater expansion of CD34+ cells and overall cell growth with a longer expansion phase, the optimal length for the expansion phase that provided greater Mk yield at near maximal purity was found to be 5 days. Under such settings, two functionally divergent cocktails were found to significantly increase the final yield of Mks. Surprisingly, these cocktails were either deprived of thrombopoietin or of stem cell factor, two cytokines known to favor megakaryopoiesis and HPC expansion, respectively. Based on these results, a short resource-efficient two-phase culture protocol for the production of Mks near purity (>95%) from human CD34+ CB cells has been established.

  12. 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%.

  13. 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