Sample records for batch culture growth

  1. [Growth and development kinetics of Bacillus thuringiensis in batch culture]. (United States)

    Sakharova, Z V; Ignatenko, Iu N; Schulz, F; Khovrychev, M P; Rabotnova, I L


    The kinetics of Bacillus thuringiensis growth and its assimilation of nutrient substances were studied under the conditions of batch cultivation in a complex medium containing yeast extract and in a chemically defined medium with amino acids. The growth of B. thuringiensis can be divided into five phases: exponential growth; decelerated growth; stationary phase when protein crystals are formed; stationary phase when spores are formed; lysis of sporangia releasing spores. The first phase may in turn be subdivided into three stages according to changes in the specific growth rate and substrate assimilation: a high specific growth rate and no glucose assimilation; an abrupt drop in mu and the beginning of intensive glucose assimilation from the medium; a new rise in the specific growth rate. As follows from the results of studying the kinetics of B. thuringiensis growth in a chemically defined medium, the above changes in the exponential growth phase are due to the fact that the culture assimilates yeast extract components in the complex medium or amino acids in the chemically defined medium during this phase, and then starts to assimilate glucose and ammonium in the following phases of growth.

  2. Modeling of Xanthophyllomyces dendrorhous growth on glucose and overflow metabolism in batch and fed-batch cultures for astaxanthin production. (United States)

    Liu, Yuan-Shuai; Wu, Jian-Yong


    An astaxanthin-producing yeast Xanthophyllomyces dendrorhous ENM5 was cultivated in a liquid medium containing 50 g/L glucose as the major carbon source in stirred fermentors (1.5-L working volume) in fully aerobic conditions. Ethanol was produced during the exponential growth phase as a result of overflow metabolism or fermentative catabolism of glucose by yeast cells. After accumulating to a peak of 3.5 g/L, the ethanol was consumed by yeast cells as a carbon source when glucose in the culture was nearly exhausted. High initial glucose concentrations and ethanol accumulation in the culture had inhibitory effects on cell growth. Astaxanthin production was partially associated with cell growth. Based on these culture characteristics, we constructed a modified Monod kinetic model incorporating substrate (glucose) and product (ethanol) inhibition to describe the relationship of cell growth rate with glucose and ethanol concentrations. This kinetic model, coupled with the Luedeking-Piret equation for the astaxanthin production, gave satisfactory prediction of the biomass production, glucose consumption, ethanol formation and consumption, and astaxanthin production in batch cultures over 25-75 g/L glucose concentration ranges. The model was also applied to fed-batch cultures to predict the optimum feeding scheme (feeding glucose and corn steep liquor) for astaxanthin production, leading to a high volumetric yield (28.6 mg/L) and a high productivity (5.36 mg/L/day).

  3. A novel approach for estimating growth phases and parameters of bacterial population in batch culture

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Huaiqiang; LIU; Yuqing; LIU; Bo; GAO; Peiji


    Using mathematical analysis, a new method has been developed for studying the growth kinetics of bacterial populations in batch culture. First, sampling data were smoothed with the spline interpolation method. Second, the instantaneous rates were derived by numerical differential techniques and finally, the derived data were fitted with the Gaussian function to obtain growth parameters. We named this the Spline-Numerical-Gaussian or SNG method. This method yielded more accurate estimates of the growth rates of bacterial populations and new parameters. It was possible to divide the growth curve into four different but continuous phases based on changes in the instantaneous rates. The four phases are the accelerating growth phase, the constant growth phase, the decelerating growth phase and the declining phase. Total DNA content was measured by flow cytometry and varied depending on the growth phase. The SNG system provides a very powerful tool for describing the kinetics of bacterial population growth. The SNG method avoids the unrealistic assumptions generally used in the traditional growth equations.

  4. Kinetic and stoichiometric characterization of organoautotrophic growth of Ralstonia eutropha on formic acid in fed-batch and continuous cultures. (United States)

    Grunwald, Stephan; Mottet, Alexis; Grousseau, Estelle; Plassmeier, Jens K; Popović, Milan K; Uribelarrea, Jean-Louis; Gorret, Nathalie; Guillouet, Stéphane E; Sinskey, Anthony


    Formic acid, acting as both carbon and energy source, is a safe alternative to a carbon dioxide, hydrogen and dioxygen mix for studying the conversion of carbon through the Calvin-Benson-Bassham (CBB) cycle into value-added chemical compounds by non-photosynthetic microorganisms. In this work, organoautotrophic growth of Ralstonia eutropha on formic acid was studied using an approach combining stoichiometric modeling and controlled cultures in bioreactors. A strain deleted of its polyhydroxyalkanoate production pathway was used in order to carry out a physiological characterization. The maximal growth yield was determined at 0.16 Cmole Cmole(-1) in a formate-limited continuous culture. The measured yield corresponded to 76% to 85% of the theoretical yield (later confirmed in pH-controlled fed-batch cultures). The stoichiometric study highlighted the imbalance between carbon and energy provided by formic acid and explained the low growth yields measured. Fed-batch cultures were also used to determine the maximum specific growth rate (μmax  = 0.18 h(-1) ) and to study the impact of increasing formic acid concentrations on growth yields. High formic acid sensitivity was found in R eutropha since a linear decrease in the biomass yield with increasing residual formic acid concentrations was observed between 0 and 1.5 g l(-1) .

  5. Growth and nutrient removal in free and immobilized green algae in batch and semi-continuous cultures treating real wastewater. (United States)

    Ruiz-Marin, Alejandro; Mendoza-Espinosa, Leopoldo G; Stephenson, Tom


    Two species of microalgae growing as immobilized and free-cells were compared to test its ability to remove N and P in batch cultures of urban wastewater. The best microalgae-cell growth configuration was selected to be tested in bioreactor operated in semi-continuous mode. Scenedesmus obliquus showed a higher N and P uptake rate in urban wastewater than Chlorella vulgaris. When tested in semi-continuous mode and with the re-calcification of beads, S. obliquus was more effective in removing N and P for longer periods (181 h) than batch cultures; fecal coliforms removal was good (95%) although the final concentration was still unsuitable for discharge to natural water bodies. Protein and lipids content analysis suggest that, from a practical point of view, immobilized systems could facilitate the separation of the biomass from the treated wastewater although in terms of nutritional value of the biomass, immobilized systems do not represent an advantage over free-cell systems.

  6. Modeling of growth and sporulation of Bacillus thuringiensis in an intermittent fed batch culture with total cell retention. (United States)

    Atehortúa, Paula; Alvarez, Hernán; Orduz, Sergio


    An extended dynamical model for growth and sporulation of Bacillus thuringiensis subsp. kurstaki in an intermittent fed-batch culture with total cell retention is proposed. This model differs from reported models, by including dynamics for natural death of cells and substrate consumption for cell maintenance. The proposed model uses sigmoid functions to describe these kinetic parameters. Equations for time evolution of substrate, vegetative, sporulated and total cell concentration were taken from previous works. Model parameters were determined from batch experimental data obtained in pilot plant. Parameter identification was developed in two stages: (1) coarse identification using a multivariable optimization with constraints algorithm, (2) fine identification by heuristic fit of model parameters looking for a minimal model error. The proposed model estimates adequate time evolution of the process variables with a mean error of 2.6% on substrate concentration and 6.7% on biomass concentration.

  7. The effect of rhizosphere on growth of Sphingomonas chlorophenolica ATCC 39723 during pentachlorophenol (PCP biodegradation in batch culture and soil

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    Ken Killhan


    Full Text Available Studies on the influence of the rhizosphere on the growth of Sphingomonas chlorophenolica during Pentacholophenol (PCP degradation in batch culture and in soil were carried out. In batch culture, a basal minimal medium with or without rhizosphere exudates extracted from winter wheat was used. In soil systems, degradation experiments were performed in the presence and absence of plants. Measurements of PCP concentrations were made using high performance liquid chromatography analysis (HPLC. Bacterial analyses of S. chlorophenolica were carried out by plating on MSM medium. The results showed that the rhizosphere exudates stimulated the growth of the cells of S. chlorophenolica at concentrations of 50 and 80mg kg dry wt soil –1 as well as stimulating the ability of S. chlorophenolica to degrade PCP at a concentration of 80mg Kg dry wt soil -1. In addition, pentachlorophenol had an adverse effect on the growth of S. chlorophenolica. The introduction of S.chlorophenolica into the loamy soil with plants showed a faster degradation when compared to the inoculated soil without plants. There was a significant increase of S. chlorophenolica in the roots in comparison to those in the soil. This study showed that the presence of the inoculum S. chlorophenolica enhanced the PCP degradation in a loamy soil and it indicates the potential for a treatment process under a appropriate environmental conditions such as there present in soil systems.

  8. Batch growth kinetics of an indigenous mixed microbial culture utilizing m-cresol as the sole carbon source. (United States)

    Saravanan, Pichiah; Pakshirajan, K; Saha, Prabirkumar


    An indigenous mixed microbial culture, isolated from a sewage treatment plant located in Guwahati was used to study biodegradation of m-cresol in batch shake flasks. m-Cresol concentration in the growth media was varied from 100mg/L to 900mg/L. The degradation kinetics was found to follow a three-half-order model at all initial m-cresol concentrations with regression values greater than 0.97. A maximum observed specific degradation rate of 0.585h(-1) was observed at 200mg/L m-cresol concentration in the medium. In the range of m-cresol concentrations used in the study, specific growth rate of the culture and specific degradation rates were observed to follow substrate inhibition kinetics. These two rates were fitted to kinetic models of Edward, Haldane, Luong, Han-Levenspiel, and Yano-Koga that are used to explain substrate inhibition on growth of microbial culture. Out of these models Luong and Han-Levenspiel models fitted the experimental data best with lowest root mean square error values. Biokinetic constants estimated from these two models showed good potential of the indigenous mixed culture in degrading m-cresol in wastewaters.

  9. Real-time monitoring and control of the specific growth rate in yeast fed-batch cultures based on process analytical technology tools such as biocalorimetry or spectroscopy


    Schuler, Moira Monika


    Key features of bioprocesses, such as product quantity and quality, but also cell physiology can be related to the growth characteristics of the organism under study. The specific growth rate, a key variable, cannot be measured directly, but might be estimated and inferred from other measurable variables such as biomass, substrate or product concentrations. The present thesis reviews techniques for real-time estimation and control of the specific growth rate in microbial fed-batch cultures by...

  10. Fermentation and growth response of a primary poultry isolate of Salmonella typhimurium grown under strict anaerobic conditions in continuous culture and amino acid-limited batch culture. (United States)

    Maciorowski, K G; Nisbet, D J; Ha, S D; Corrier, D E; DeLoach, J R; Ricke, S C


    Salmonella typhimurium is a significant hazard to consumer health that is carried asymptomatically in poultry gastrointestinal tracts. Nurmi cultures may prevent Salmonella colonization in young chicks, but the mechanism of competitive exclusion is unclear. Modeling Salmonella's metabolism in pure culture may allow for greater definition in choosing strains for Nurmi cultures. The growth rates and affinity constants of S. typhimurium growing in amino acid-limited conditions were determined in batch culture and compared to primary poultry isolates of cecal strains. Serine and NH4Cl were the best N sources for growth of all organisms tested in this study. The fermentation response of S. typhimurium was also monitored in continuous culture at a slow dilution rate of 0.021 h-1. S. typhimurium was found to adapt to VL media, with trends in protein disappearance, Yglucose, and Yprotein. This may show that amino acid or protein concentrations may be an integral component of the initial establishment of S. typhimurium in the cecum.

  11. Probabilistic neural networks using Bayesian decision strategies and a modified Gompertz model for growth phase classification in the batch culture of Bacillus subtilis. (United States)

    Simon; Nazmul Karim M


    Probabilistic neural networks (PNNs) were used in conjunction with the Gompertz model for bacterial growth to classify the lag, logarithmic, and stationary phases in a batch process. Using the fermentation time and the optical density of diluted cell suspensions, sampled from a culture of Bacillus subtilis, PNNs enabled a reliable determination of the growth phases. Based on a Bayesian decision strategy, the Gompertz based PNN used newly proposed definition of the lag and logarithmic phases to estimate the latent, logarithmic and stationary phases. This network topology has the potential for use with on-line turbidimeter for the automation and control of cultivation processes.

  12. Endoplasmic Reticulum-Associated rht-PA Processing in CHO Cells: Influence of Mild Hypothermia and Specific Growth Rates in Batch and Chemostat Cultures.

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    Mauricio Vergara

    Full Text Available Chinese hamster ovary (CHO cells are the main host for producing recombinant proteins with human therapeutic applications mainly because of their capability to perform proper folding and glycosylation processes. In addition, mild hypothermia is one of the main strategies for maximising the productivity of these systems. However, little information is available on the effect of culture temperature on the folding and degradation processes of recombinant proteins that takes place in the endoplasmic reticulum.In order to evaluate the effect of the mild hypothermia on processing/endoplasmatic reticulum-associated degradation (ERAD processes, batch cultures of CHO cells producing recombinant human tissue plasminogen activator (rht-PA were carried out at two temperatures (37°C and 33°C and treated with specific inhibitors of glycosylation and ERAD I (Ubiquitin/Proteasome system or ERAD II (Autophagosoma/Lisosomal system pathways. The effect of mild hypothermia was analysed separately from its indirect effect on specific cell growth rate. To do this, chemostat cultures were carried out at the same incubation conditions as the batch cultures, controlling cell growth at high (0.017 h-1 and low (0.012 h-1 dilution rates. For a better understanding of the investigated phenomenon, cell behaviour was also analysed using principal component analysis (PCA.Results suggest that rht-PA is susceptible to degradation by both ERAD pathways studied, revealing that processing and/or ERAD processes are sensitive to temperature cultivation in batch culture. Moreover, by isolating the effect of culture temperature from the effect of cell growth rate verifyed by using chemostat cultures, we have found that processing and/or ERAD processes are more sensitive to reduction in specific growth rate than low temperature, and that temperature reduction may have a positive effect on protein processing. Interestingly, PCA indicated that the integrated performance displayed by CHO

  13. In vitro growth of Curcuma longa L. in response to five mineral elements and plant density in fed-batch culture systems. (United States)

    El-Hawaz, Rabia F; Bridges, William C; Adelberg, Jeffrey W


    Plant density was varied with P, Ca, Mg, and KNO3 in a multifactor experiment to improve Curcuma longa L. micropropagation, biomass and microrhizome development in fed-batch liquid culture. The experiment had two paired D-optimal designs, testing sucrose fed-batch and nutrient sucrose fed-batch techniques. When sucrose became depleted, volume was restored to 5% m/v sucrose in 200 ml of modified liquid MS medium by adding sucrose solutions. Similarly, nutrient sucrose fed-batch was restored to set points with double concentration of treatments' macronutrient and MS micronutrient solutions, along with sucrose solutions. Changes in the amounts of water and sucrose supplementations were driven by the interaction of P and KNO3 concentrations. Increasing P from 1.25 to 6.25 mM increased both multiplication and biomass. The multiplication ratio was greatest in the nutrient sucrose fed-batch technique with the highest level of P, 6 buds/vessel, and the lowest level of Ca and KNO3. The highest density (18 buds/vessel) produced the highest fresh biomass at the highest concentrations of KNO3 and P with nutrient sucrose fed-batch, and moderate Ca and Mg concentrations. However, maximal rhizome dry biomass required highest P, sucrose fed-batch, and a moderate plant density. Different media formulations and fed-batch techniques were identified to maximize the propagation and storage organ responses. A single experimental design was used to optimize these dual purposes.

  14. Enhanced growth and recombinant protein production of Escherichia coli by a perfluorinated oxygen carrier in miniaturized fed-batch cultures

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    Neubauer Peter


    Full Text Available Abstract Background Liquid perfluorochemicals (PFCs are interesting oxygen carriers in medicine and biotechnology with a high solubility for oxygen. They have been repeatedly used for improving oxygen transfer into prokaryotic and eukaryotic cell cultures, however their application is still limited. Here we show the great benefit of air/oxygen saturated perfluorodecalin (PFD for high cell density cultivation of Escherichia coli in microwell plates and their positive effect on the soluble production of a correctly folded heterologously expressed alcohol dehydrogenase. Results In EnBase® cultivations the best effect was seen with PFD saturated with oxygen enriched air (appr. 10 μM oxygen per ml when PFD was added at the time of induction. In contrast the effect of PFD was negligible when it was added already at the time of inoculation. Optimisation of addition time and content of loaded oxygen into the PFD resulted in an increased the cell density by 40% compared to control cultures, and correspondingly also the product yield increased, demonstrated at the example of a recombinant alcohol dehydrogenase. Conclusions PFCs are a valuable additive in miniaturized cell culture formats. For production of recombinant proteins in low cell density shaken cultures the addition of oxygen-enriched PFD makes the process more robust, i.e. a high product yield is not any more limited to a very narrow cell density window during which the induction has to be done. The positive effect of PFD was even more obvious when it was added during high cell density cultures. The effect of the PFD phase depends on the amount of oxygen which is loaded into the PFD and which thus is a matter of optimisation.

  15. Application of data consistency tests and new parameter estimation methods to microbial growth on corn dust in batch culture

    Energy Technology Data Exchange (ETDEWEB)

    Solomon, B.O.; Erickson, L.E.; Hess, J.L.


    The microorganism Candida utilis was grown on both filtered and unfiltered substrate obtained from enzymatic hydrolysis of starch in corn dust. For growth on filtered substrate, the average integrated biomass energetic yield value based on biomass-substrate data was eta = 0.55 and for growth on unfiltered substrate an average yield value of eta = 0.59 was obtained. Material and energy balances showed that the presence of unfiltered corn residue in the media had no significant effect on the yields. Statistical methods were developed and used to obtain best estimates of the growth parameters. Values of the biomass energetic yield corrected for maintenance (eta maximum = 0.619) and the maintenance coefficient (Me = 0.043) were obtained for growth on filtered substrate. Values of the maximum = 0.741 and Me = 0.142 were obtained for the grown on unfiltered substrate. The consistency of data and parameter estimates was relatively good for filtered substrate; however, the parameter estimates for unfiltered substrate were not consistent. Growth experiments without filtration of the products of starch hydrolysis resulted in protein-enriched products with about 39.73% protein. (Refs. 26).

  16. Growth and nitrogen removal capacity of Desmodesmus communis and of a natural microalgae consortium in a batch culture system in view of urban wastewater treatment: part I. (United States)

    Samorì, Giulia; Samorì, Chiara; Guerrini, Franca; Pistocchi, Rossella


    The microalgal biomass applications strongly depend on cell composition and the production of low cost products such as biofuels appears to be economically convenient only in conjunction with wastewater treatment. As a preliminary study, in view of the development of a wastewater treatment pilot plant for nutrient removal and algal biomass production, a biological wastewater system was carried out on a laboratory scale growing a newly isolated freshwater algal strain, Desmodesmus communis, and a natural consortium of microalgae in effluents generated by a local wastewater reclamation facility. Batch cultures were operated by using D. communis under different growth conditions to better understand the effects of CO₂, nutrient concentration and light intensity on the biomass productivity and biochemical composition. The results were compared with those obtained using a natural algal consortium. D. communis showed a great vitality in the wastewater effluents with a biomass productivity of 0.138-0.227 g L⁻¹ d⁻¹ in the primary effluent enriched with CO₂, higher biomass productivity compared with the one achieved by the algal consortium (0.078 g L⁻¹ d⁻¹). D. communis cultures reached also a better nutrient removal efficiency compared with the algal consortium culture, with almost 100% for ammonia and phosphorous at any N/P ratio characterizing the wastewater nutrient composition. Biomass composition was richer in polysaccharides and total fatty acids as the ammonia concentration in the water decreased. In view of a future application of this algal biomass, due to the low total fatty acids content of 1.4-9.3 wt% and the high C/N ratio of 7.6-39.3, anaerobic digestion appeared to be the most appropriate biofuel conversion process.

  17. Evaluation of cyanobacteria: Spirulina maxima for growth, nutrient removal, and quality on waste-effluent media in batch cultures

    Energy Technology Data Exchange (ETDEWEB)

    Tadros, M.G.; Phillips, J. (Alabama A M Univ., Normal (United States))


    Spirulina maxima, a semi-microscopic filamentous blue-green alga, was inoculated in synthetic and waste media of different sources. The alga was evaluated for growth yield, uptake of nutrients and chemical composition. The removal rate of N and P was rapid during the first week of growth. At the end of the second week, more than 90% of the total -P and -N was removed. The mass of alga was high. The quality of the alga obtained in different media did not show much variations, except when the medium was limited in nutrients. Results indicated that Spirulina may be integrated into the effluent treatment system. Recycling waste materials not only minimizes the problem of water pollution but also revitalizes the inherently rich nutrients of waste. The biomass obtained from cultivation of Spirulina in these wastewater media may be used as a pigment-protein supplement in animal feed and as raw material for certain chemicals.

  18. Comprehensive modeling and investigation of the effect of iron on the growth rate and lipid accumulation of Chlorella vulgaris cultured in batch photobioreactors. (United States)

    Concas, Alessandro; Steriti, Alberto; Pisu, Massimo; Cao, Giacomo


    Recent works have shown that specific strains of microalgae are capable to simultaneously increase their growth rate and lipid content when cultured under suitable concentrations of iron. While these results are promising in view of the exploitation of microalgae for producing biofuels, to the best of our knowledge, no mathematical model capable to describe the effect of iron on lipid accumulation in microalgae, has been so far proposed. A comprehensive mathematical model describing the effect of iron on chlorophyll synthesis, nitrogen assimilation, growth rate and lipid accumulation in a freshwater strain of Chlorella vulgaris is then proposed in this work. Model results are successfully compared with experimental data which confirm the positive effect of growing iron concentrations on lipid productivity of C. vulgaris. Thus, the proposed model might represent a useful tool to optimize iron-based strategies to improve the lipid productivity of microalgal cultures.

  19. Batch variation between branchial cell cultures: An analysis of variance

    DEFF Research Database (Denmark)

    Hansen, Heinz Johs. Max; Grosell, M.; Kristensen, L.


    We present in detail how a statistical analysis of variance (ANOVA) is used to sort out the effect of an unexpected batch-to-batch variation between cell cultures. Two separate cultures of rainbow trout branchial cells were grown on permeable filtersupports ("inserts"). They were supposed...... and introducing the observed difference between batches as one of the factors in an expanded three-dimensional ANOVA, we were able to overcome an otherwisecrucial lack of sufficiently reproducible duplicate values. We could thereby show that the effect of changing the apical medium was much more marked when...... the radioactive lipid precursors were added on the apical, rather than on the basolateral, side. Theinsert cell cultures were obviously polarized. We argue that it is not reasonable to reject troublesome experimental results, when we do not know a priori that something went wrong. The ANOVA is a very useful...

  20. Stochastic growth logistic model with aftereffect for batch fermentation process

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    Rosli, Norhayati; Ayoubi, Tawfiqullah [Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, Lebuhraya Tun Razak, 26300 Gambang, Pahang (Malaysia); Bahar, Arifah; Rahman, Haliza Abdul [Department of Mathematical Sciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Salleh, Madihah Md [Department of Biotechnology Industry, Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia)


    In this paper, the stochastic growth logistic model with aftereffect for the cell growth of C. acetobutylicum P262 and Luedeking-Piret equations for solvent production in batch fermentation system is introduced. The parameters values of the mathematical models are estimated via Levenberg-Marquardt optimization method of non-linear least squares. We apply Milstein scheme for solving the stochastic models numerically. The effciency of mathematical models is measured by comparing the simulated result and the experimental data of the microbial growth and solvent production in batch system. Low values of Root Mean-Square Error (RMSE) of stochastic models with aftereffect indicate good fits.

  1. Stochastic growth logistic model with aftereffect for batch fermentation process (United States)

    Rosli, Norhayati; Ayoubi, Tawfiqullah; Bahar, Arifah; Rahman, Haliza Abdul; Salleh, Madihah Md


    In this paper, the stochastic growth logistic model with aftereffect for the cell growth of C. acetobutylicum P262 and Luedeking-Piret equations for solvent production in batch fermentation system is introduced. The parameters values of the mathematical models are estimated via Levenberg-Marquardt optimization method of non-linear least squares. We apply Milstein scheme for solving the stochastic models numerically. The effciency of mathematical models is measured by comparing the simulated result and the experimental data of the microbial growth and solvent production in batch system. Low values of Root Mean-Square Error (RMSE) of stochastic models with aftereffect indicate good fits.

  2. Heterotrophic high cell-density fed-batch cultures of the phycocyanin-producing red alga Galdieria sulphuraria. (United States)

    Schmidt, Rikke Ankerstjerne; Wiebe, Marilyn G; Eriksen, Niels Thomas


    Growth and phycocyanin production in batch and fed-batch cultures of the microalga Galdieria sulphuraria 074G, which was grown heterotrophically in darkness on glucose, fructose, sucrose, and sugar beet molasses, was investigated. In batch cultures, specific growth rates and yields of biomass dry weight on the pure sugars were 1.08-1.15 day-1 and 0.48-0.50 g g-1, respectively. They were slightly higher when molasses was the carbon source. Cellular phycocyanin contents during the exponential growth phase were 3-4 mg g-1 in dry weight. G. sulphuraria was able to tolerate concentrations of glucose and fructose of up to 166 g L-1 (0.9 M) and an ammonium sulfate concentration of 22 g L-1 (0.17 M) without negative effects on the specific growth rate. When the total concentration of dissolved substances in the growth medium exceeded 1-2 M, growth was completely inhibited. In carbon-limited fed-batch cultures, biomass dry weight concentrations of 80-120 g L-1 were obtained while phycocyanin accumulated to concentrations between 250 and 400 mg L-1. These results demonstrate that G. sulphuraria is well suited for growth in heterotrophic cultures at very high cell densities, and that such cultures produce significant amounts of phycocyanin. Furthermore, the productivity of phycocyanin in the heterotrophic fed-batch cultures of G. sulphuraria was higher than is attained in outdoor cultures of Spirulina platensis, where phycocyanin is presently obtained.

  3. Biodegradation of phenanthrene in an anaerobic batch reactor: growth kinetics

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    H.S. Nasrollahzadeh


    Full Text Available The purpose of the present research was to demonstrate the ability of mixed consortia of microorganisms to degrade high concentrations of phenanthrene (PHE as the sole carbon source. Batch experiments were carried out by the induction of mineral salt medium containing PHE to the seed culture and monitoring PHE biodegradation. The microbial propagation was conducted using PHE concentrations in the range of 20 to 100 mg/l. The microbial growth on PHE was defined based on Monod and modified Logistic rate models. The kinetic studies revealed that maximum specific growth rates (μm for PHE concentrations of 20, 50 and 100 mg/l were 0.12, 0.23 and 0.035 h-1, respectively. The doubling times for microbial population in PHE concentrations of 20, 50 and 100 mg/l were 13, 15 and 17.5 h, respectively. Also, maximum cell dry weight (xm of 54.23 mg/l was achieved, while the inhibition coefficient was 0.023 h-1. It was observed that the experimental data were well represented by the proposed models. It was also found that the biodegradation of PHE was successfully performed by the isolated strains.

  4. Oxygen availability strongly affects chronological lifespan and thermotolerance in batch cultures of Saccharomyces cerevisiae

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    Markus M.M. Bisschops


    Full Text Available Stationary-phase (SP batch cultures of Saccharomyces cerevisiae, in which growth has been arrested by carbon-source depletion, are widely applied to study chronological lifespan, quiescence and SP-associated robustness. Based on this type of experiments, typically performed under aerobic conditions, several roles of oxygen in aging have been proposed. However, SP in anaerobic yeast cultures has not been investigated in detail. Here, we use the unique capability of S. cerevisiae to grow in the complete absence of oxygen to directly compare SP in aerobic and anaerobic bioreactor cultures. This comparison revealed strong positive effects of oxygen availability on adenylate energy charge, longevity and thermotolerance during SP. A low thermotolerance of anaerobic batch cultures was already evident during the exponential growth phase and, in contrast to the situation in aerobic cultures, was not substantially increased during transition into SP. A combination of physiological and transcriptome analysis showed that the slow post-diauxic growth phase on ethanol, which precedes SP in aerobic, but not in anaerobic cultures, endowed cells with the time and resources needed for inducing longevity and thermotolerance. When combined with literature data on acquisition of longevity and thermotolerance in retentostat cultures, the present study indicates that the fast transition from glucose excess to SP in anaerobic cultures precludes acquisition of longevity and thermotolerance. Moreover, this study demonstrates the importance of a preceding, calorie-restricted conditioning phase in the acquisition of longevity and stress tolerance in SP yeast cultures, irrespective of oxygen availability.

  5. Improving the batch-to-batch reproducibility of microbial cultures during recombinant protein production by regulation of the total carbon dioxide production. (United States)

    Jenzsch, Marco; Gnoth, Stefan; Kleinschmidt, Martin; Simutis, Rimvydas; Lübbert, Andreas


    Batch-to-batch reproducibility of fermentation processes performed during the manufacturing processes of biologics can be increased by operating the cultures at feed rate profiles that are robust against typically arising disturbances. Remaining randomly appearing deviations from the desired path should be suppressed automatically by manipulating the feed rate. With respect to the cells' physiology it is best guiding the cultivations along an optimal profile of the specific biomass growth rate mu(t). However, there are two problems that speak for further investigations: Upon severe disturbances that may happen during the fermentation, the biomass concentration X may significantly deviate from its desired value, then a fixed mu-profile leads to a diminished batch-to-batch reproducibility. Second, the specific growth rate cannot easily be estimated online to a favourably high accuracy, hence it is difficult to determine the deviations in mu from the desired profile. The alternative discussed here solves both problems by keeping the process at the corresponding total cumulative carbon dioxide production-profile: it is robust against distortions in X and the controlled variable can accurately be measured online during cultivations of all relevant sizes. As compared to the fermentation practice currently used in industry, the experimental results, presented at the example of a recombinant protein production with Escherichia coli cells, show that CPR-based corrections lead to a considerably improved batch-to-batch reproducibility.

  6. Growth kinetics of Saccharomyces cerevisiae in batch and fed-batch cultivation using sugarcane molasses and glucose syrup from cassava starch. (United States)

    Win, S S; Impoolsup, A; Noomhorm, A


    Growth kinetics of Saccharomyces cerevisiae in glucose syrup from cassava starch and sugarcane molasses were studied using batch and fed-batch cultivation. The optimum temperature and pH required for growth were 30 degrees C and pH 5.5, respectively. In batch culture the productivity and overall cell yield were 0.31 g L-1 h-1 and 0.23 g cells g-1 sugar, respectively, on glucose syrup and 0.22 g L-1 h-1 and 0.18 g cells g-1 sugar, respectively, on molasses. In fed-batch cultivation, a productivity of 3.12 g L-1 h-1 and an overall cell yield of 0.52 g cells g-1 sugar were achieved in glucose syrup cultivation and a productivity of 2.33 g L-1 h-1 and an overall cell yield of 0.46 g cells g-1 sugar were achieved in molasses cultivation by controlling the reducing sugar concentration at its optimum level obtained from the fermentation model. By using an on-line ethanol sensor combined with a porous Teflon tubing method in automating the feeding of substrate in the fed-batch culture, a productivity of 2.15 g L-1 h-1 with a yield of 0.47 g cells g-1 sugar was achieved using glucose syrup as substrate when ethanol concentration was kept at a constant level by automatic control.

  7. On-line optimal control for fed-batch culture of baker's yeast production

    Energy Technology Data Exchange (ETDEWEB)

    Wu, W.T.; Chen, K.C.; Chiou, H.W.


    A method of on-line optimal control for fed-batch culture of bakers yeast production is proposed. The feed rate is taken as the control variable. The specific growth rate of the yeast is the output variable and is determined from the balance equation of oxygen. A moving model is obtained by using the data from the feed rate and the specific growth rate. Based on the moving model, an optimal feed rate for fed-batch culture is then achieved. 11 references.

  8. Photosynthetic light reactions increase total lipid accumulation in carbon-supplemented batch cultures of Chlorella vulgaris. (United States)

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


    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).

  9. Behavior of transition state regulator AbrB in batch cultures of Bacillus thuringiensis. (United States)

    Lozano Goné, Astrid Magdalena; Dinorín Téllez Girón, Jabel; Jiménez Montejo, Fabiola Eloisa; Hidalgo-Lara, María Eugenia; López Y López, Víctor Eric


    The transition state regulator AbrB is involved in the regulation of various cellular functions such as exponential growth, transition state and sporulation onset, due to its ability to activate, suppress or prevent the inappropriate expression of various genes in Bacillus subtilis. In order to understand combined behavior in batch cultures of AbrB in Bacillus thuringiensis, we cloned and expressed the abrB gene of B. thuringiensis in Escherichia coli. The deduced sequence of abrB gene coded for a protein consisting of 94 amino acids with ~10.5 kDa protein that shares 100 and 85 % identity with those from Bacillus cereus and Bacillus subtilis. The recombinant AbrB protein was used as antigen for the production of rabbit polyclonal antibodies anti-AbrB. Two media cultures with carbon: nitrogen ratios of 7.0, but varying access to nutrients were tested in batch cultures. In the case of both media, AbrB accumulation occurred from the beginning of the process and was maximal during early exponential growth. Thereafter, the level of AbrB decreased when there were no nutrient limitations and coincided with a decreased value in specific growth rate, although growth continued exponentially. Nonetheless, sporulation onset was determined 3 h and 4 h later, in media with highly metabolizable nutrients clean medium and Farrera medium, respectively. Hence, the maximal level of AbrB accumulation in batch cultures of B. thuringiensis is not influenced by limiting nutrients; however, nutrient availability affects the required time lapse for transition state regulator accumulation.

  10. Biodesulfurization of Malaysian coals using mixed microbial cultures in batch and continuous processes

    Energy Technology Data Exchange (ETDEWEB)

    Najafpour, G.D.; Azizan, A.; Harun, A. [Universiti Sains Malaysia, Darul Ridzuan (Malaysia). School of Chemical Engineering


    The determination of chemolithotrophic microorganisms capable to grown on coal with high sulfur content, made it possible to develop mixed culture processes for coal desulfurization. The ability of the microbial cultures to metabolize different sulfur compounds originated from coal in the range of 3-5% sulfur, were demonstrated in batch experimental stage. Biodesulfurization of coal as suspended solid coal particles were carried out in a broth media for coal content of 1-5% solid. Four species of microbial culture were grown on coal and acclimated for optimal growth. The coal samples obtained from Malaysian coal mine with 2-5% of sulfur content were used in batch and continuous cultures experiments. The microbial cultures were used to reduce pyrite sulfur, inorganic sulfur content of coal. The culture isolated from pharmaceutical wastewater shown that more than 80% of sulfur content of coal was reduced. The growth of microorganisms on coal shown that maximum solid content of 5% was tolerable maximum grow on 3% coal shown high cell density. Nutrient media with acidic PH value of 3-4 was required for growth of Thiobacillus thiooxidans and Thiobacillus fierrooxidans. The growth was limited at PH = 2 but at PH 3-4 growth was stimulated. Phosphate buffer was used with coal solution to monitor the PH. The PH was initially adjusted but was not controlled during the period of incubation. An optimum PH for Thiobacillus species with maximum desulfuriation was 2.65. High coal content about 10% was shown that bacterial was unable to grow. The cell dry weight, cell optical density and sulfur content of coal samples using TGA and elemental analyzer were conducted for all experiments. 10 refs., 11 figs.

  11. Algae for controlled ecological life support system diet characterization of cyanobacteria 'spirulina' in batch cultures (United States)

    Tadros, M. G.


    Spirulina sp. is a bioregenerative photosynthetic and edible alga for space craft crews in a Closed Ecological Life Support System (CLESS). It was characterized for growth rate and biomass yield in batch cultures, under various environmental conditions. The cell characteristics were identified for one strain of Spirulina: S. maxima. Fast growth rate and high yield were obtained. The partitioning of the assimulatory products (proteins, carbohydrates, lipids) were manipulated by varying the environmental conditions. Experiments with Spirulina demonstrated that under stress conditions carbohydrate increased at the expense of protein. In other experiments, where the growth media were sufficient in nutrients and incubated under optimum growth conditions, the total proteins were increased up to almost 70 percent of the organic weight. In other words, the nutritional quality of the alga could be manipulated by growth conditions. These results support the feasibility of considering Spirulina as a subsystem in CELSS because of the ease with which its nutrient content can be manipulated.


    Directory of Open Access Journals (Sweden)

    Leonard D. Holmes


    Full Text Available The present study deals with the batch and fed-batch mass production of Steinernema carpocapsae. S. carpocapsae is an entomoparasitic nematode that is used as a biological control agent of soil-borne crop insect pests. The ability and efficiency of fed-batch culture process was successful through the utilization of the nematode’s bacterial symbiont Xenorhabdus nematophila. Results from the fed-batch process were compared to those obtain from the standard batch process. The fed-batch process successively improved the mass production process of S. carpocapsae employing liquid medium technology. Within the first week of the fed-batch process (day six, the nematode density obtained was 202,000 nematodes mL−1; whereas on day six, batch culture mode resulted in a nematode density of 23,000 nematodes mL−1. The fed-batch process was superior to that of batch production with a yield approximately 8.8-fold higher. In fed-batch process, the nematode yield was improved 88.6 % higher within a short amount of time compared to the batch process. Fed-batch seems to make the process more efficient and possibly economically viable.

  13. Effect of Selenate on Viability and Selenomethionine Accumulation of Chlorella sorokiniana Grown in Batch Culture

    Directory of Open Access Journals (Sweden)

    Živan Gojkovic


    Full Text Available The aim of this work was to study the effect of Se(+VI on viability, cell morphology, and selenomethionine accumulation of the green alga Chlorella sorokiniana grown in batch cultures. Culture exposed to sublethal Se concentrations of 40 mg·L−1 (212 μM decreased growth rates for about 25% compared to control. A selenate EC50 value of 45 mg·L−1 (238.2 μM was determined. Results showed that chlorophyll and carotenoids contents were not affected by Se exposure, while oxygen evolution decreased by half. Ultrastructural studies revealed granular stroma, fingerprint-like appearance of thylakoids which did not compromise cell activity. Unlike control cultures, SDS PAGE electrophoresis of crude extracts from selenate-exposed cell cultures revealed appearance of a protein band identified as 53 kDa Rubisco large subunit of Chlorella sorokiniana, suggesting that selenate affects expression of the corresponding chloroplast gene as this subunit is encoded in the chloroplast DNA. Results revealed that the microalga was able to accumulate up to 140 mg·kg−1 of SeMet in 120 h of cultivation. This paper shows that Chlorella sorokiniana biomass can be enriched in the high value aminoacid SeMet in batch cultures, while keeping photochemical viability and carbon dioxide fixation activity intact, if exposed to suitable sublethal concentrations of Se.

  14. Segmented linear modeling of CHO fed‐batch culture and its application to large scale production (United States)

    Ben Yahia, Bassem; Gourevitch, Boris; Malphettes, Laetitia


    ABSTRACT We describe a systematic approach to model CHO metabolism during biopharmaceutical production across a wide range of cell culture conditions. To this end, we applied the metabolic steady state concept. We analyzed and modeled the production rates of metabolites as a function of the specific growth rate. First, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. For this, the smoothed derivative of the metabolic rates with respect to the growth rate were used followed by hierarchical clustering of the obtained partition. We then applied a piecewise regression to the metabolic rates with the previously determined number of phases. This allowed identifying the growth rates at which the cells underwent a metabolic shift. The resulting model with piecewise linear relationships between metabolic rates and the growth rate did well describe cellular metabolism in the fed‐batch cultures. Using the model structure and parameter values from a small‐scale cell culture (2 L) training dataset, it was possible to predict metabolic rates of new fed‐batch cultures just using the experimental specific growth rates. Such prediction was successful both at the laboratory scale with 2 L bioreactors but also at the production scale of 2000 L. This type of modeling provides a flexible framework to set a solid foundation for metabolic flux analysis and mechanistic type of modeling. Biotechnol. Bioeng. 2017;114: 785–797. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. PMID:27869296

  15. Lipid production in batch and fed-batch cultures of Rhodosporidium toruloides from 5 and 6 carbon carbohydrates

    Directory of Open Access Journals (Sweden)

    Wiebe Marilyn G


    Full Text Available Abstract Background Microbial lipids are a potential source of bio- or renewable diesel and the red yeast Rhodosporidium toruloides is interesting not only because it can accumulate over 50% of its dry biomass as lipid, but also because it utilises both five and six carbon carbohydrates, which are present in plant biomass hydrolysates. Methods R. toruloides was grown in batch and fed-batch cultures in 0.5 L bioreactors at pH 4 in chemically defined, nitrogen restricted (C/N 40 to 100 media containing glucose, xylose, arabinose, or all three carbohydrates as carbon source. Lipid was extracted from the biomass using chloroform-methanol, measured gravimetrically and analysed by GC. Results Lipid production was most efficient with glucose (up to 25 g lipid L−1, 48 to 75% lipid in the biomass, at up to 0.21 g lipid L−1 h−1 as the sole carbon source, but high lipid concentrations were also produced from xylose (36 to 45% lipid in biomass. Lipid production was low (15–19% lipid in biomass with arabinose as sole carbon source and was lower than expected (30% lipid in biomass when glucose, xylose and arabinose were provided simultaneously. The presence of arabinose and/or xylose in the medium increased the proportion of palmitic and linoleic acid and reduced the proportion of oleic acid in the fatty acids, compared to glucose-grown cells. High cell densities were obtained in both batch (37 g L−1, with 49% lipid in the biomass and fed-batch (35 to 47 g L−1, with 50 to 75% lipid in the biomass cultures. The highest proportion of lipid in the biomass was observed in cultures given nitrogen during the batch phase but none with the feed. However, carbohydrate consumption was incomplete when the feed did not contain nitrogen and the highest total lipid and best substrate consumption were observed in cultures which received a constant low nitrogen supply. Conclusions Lipid production in R. toruloides was lower from arabinose and mixed

  16. Macroscopic modelling of bioethanol production from potato peel wastes in batch cultures supplemented with inorganic nitrogen. (United States)

    Richelle, A; Ben Tahar, I; Hassouna, M; Bogaerts, Ph


    Inorganic nitrogen supplementation is commonly used to boost fermentation metabolism in yeast cultures. However, an excessive addition can induce an opposite effect. Hence, it is important to ensure that the ammonia supplemented to the culture leads to an improvement of the ethanol production while avoiding undesirable inhibition effects. To this end, a macroscopic model describing the influence of ammonia addition on Saccharomyces cerevisiae metabolism during bioethanol production from potato peel wastes has been developed. The model parameters are obtained by a simplified identification methodology in five steps. It is validated with experimental data and successfully predicts the dynamics of growth, substrate consumption (ammonia and fermentable sugar sources) and bioethanol production, even in cross validation. The model is used to determine the optimal quantity of supplemented ammonia required for maximizing bioethanol production from potato peel wastes in batch cultures.

  17. A Single Dynamic Metabolic Model Can Describe mAb Producing CHO Cell Batch and Fed-Batch Cultures on Different Culture Media. (United States)

    Robitaille, Julien; Chen, Jingkui; Jolicoeur, Mario


    CHO cell culture high productivity relies on optimized culture medium management under fed-batch or perfused chemostat strategies enabling high cell densities. In this work, a dynamic metabolic model for CHO cells was further developed, calibrated and challenged using datasets obtained under four different culture conditions, including two batch and two fed-batch cultures comparing two different culture media. The recombinant CHO-DXB11 cell line producing the EG2-hFc monoclonal antibody was studied. Quantification of extracellular substrates and metabolites concentration, viable cell density, monoclonal antibody concentration and intracellular concentration of metabolite intermediates of glycolysis, pentose-phosphate and TCA cycle, as well as of energetic nucleotides, were obtained for model calibration. Results suggest that a single model structure with a single set of kinetic parameter values is efficient at simulating viable cell behavior in all cases under study, estimating the time course of measured and non-measured intracellular and extracellular metabolites. Model simulations also allowed performing dynamic metabolic flux analysis, showing that the culture media and the fed-batch strategies tested had little impact on flux distribution. This work thus paves the way to an in silico platform allowing to assess the performance of different culture media and fed-batch strategies.

  18. Modelling of Escherichia coli Cultivations: Acetate Inhibition in a Fed-batch Culture

    Directory of Open Access Journals (Sweden)

    Stoyan Tzonkov


    Full Text Available A set of three competing, unstructured models has been proposed to model biomass growth, glucose utilization, acetate formation, dissolved oxygen consumption and carbon dioxide accumulation of a fed-batch cultivation process of Escherichia coli. The inhibiting effect of acetate on growth of E. coli cultures is included in the considered models. The model identification is carried out using experimental data from the cultivation process. Genetic algorithms are used for parameter estimation. The model discrimination is based on the four criteria, namely sum of square errors, Fisher criterion, Akaike information criterion and minimum description length criterion. The most suitable model is identified that reflects the state variables curves adequately by considering acetate inhibited growth according to the Jerusalimsky approach.

  19. Production of cellulase using a mutant strain of trichoderma reesei growing on lactose in batch culture

    Energy Technology Data Exchange (ETDEWEB)

    Chaudhuri, B.K. (Indian Inst. of Tech., New Delhi (India). Biochemical Engineering Research Centre); Sahai, V. (Indian Inst. of Tech., New Delhi (India). Biochemical Engineering Research Centre)


    The production of cellulases in batch culture was studied using a mutant strain of Trichoderma reesei C-5 growing on lactose. Growth kinetic parameters on 2% lactose were studied and the comparative results for growth and enzyme productivities at two different lactose levels are discussed. The cellulase synthesis rate depended on the lactose concentration in the medium. Although growth was favoured at a higher lactose level, the volumetric enzyme productivity did not increase in proportion and the specific enzyme productivity decreased to a certain extent, indicating that partial catabolic inhibition at higher lactose concentrations may be possible. However, it was noted that the mutant strain was highly derepressed and capable of synthesising active cellulases on lactose. (orig.)

  20. Nisin production in realkalized fed-batch cultures in whey with feeding with lactose- or glucose-containing substrates. (United States)

    Costas Malvido, Mónica; Alonso González, Elisa; Pérez Guerra, Nelson


    Nisin production by Lactococcus lactis CECT 539 was followed in batch cultures in whey supplemented with different concentrations of glucose and in two realkalized fed-batch fermentations in unsupplemented whey, which were fed, respectively, with concentrated solutions of lactose and glucose. In the batch fermentations, supplementation of whey with glucose inhibited both the growth and bacteriocin production. However, fed-batch cultures were characterized with high productions of biomass (1.34 and 1.51 g l(-1)) and nisin (50.6 and 60.3 BU ml(-1)) in comparison to the batch fermentations in unsupplemented whey (0.48 g l(-1) and 22.5 BU ml(-1)) and MRS broth (1.59 g l(-1) and 50.0 BU ml(-1)). In the two realkalized fed-batch fermentations, the increase in bacteriocin production parallels both the biomass production and pH drop generated in each realkalization and feeding cycle, suggesting that nisin was synthesized as a pH-dependent primary metabolite. A shift from homolactic to heterolactic fermentation was observed at the 108 h of incubation, and other metabolites (acetic acid and butane-2,3-diol) in addition to lactic acid accumulated in the medium. On the other hand, the feeding with glucose improved the efficiencies in glucose, nitrogen, and phosphorus consumption as compared to the batch cultures. The realkalized fed-batch fermentations showed to be an effective strategy to enhance nisin production in whey by using an appropriate feeding strategy to avoid the substrate inhibition.

  1. Modelling of hydrogen production in batch cultures of the photosynthetic bacterium Rhodobacter capsulatus

    Energy Technology Data Exchange (ETDEWEB)

    Obeid, Jamila; Magnin, Jean-Pierre [Grenoble Institute of Technology, LEPMI, UMR 5631 (CNRS-INPG-UJF), BP 75, 38402 St Martin d' Heres (France); Flaus, Jean-Marie; Adrot, Olivier [Grenoble Institute of Technology, Laboratoire des sciences pour la conception, l' optimisation et la production, 46, avenue Felix Viallet, 38031 Grenoble (France); Willison, John C. [Laboratoire de Chimie et Biologie des Metaux (UMR 5249 CEA-CNRS-UJF), iRTSV/LCBM, CEA-Grenoble, 38054 Grenoble (France); Zlatev, Roumen [Autonomous University of Baja California, Institute of Engineering, Mexicali, Baja California (Mexico)


    The photosynthetic bacterium, Rhodobacter capsulatus, produces hydrogen under nitrogen-limited, anaerobic, photosynthetic culture conditions, using various carbon substrates. In the present study, the relationship between light intensity and hydrogen production has been modelled in order to predict both the rate of hydrogen production and the amount of hydrogen produced at a given time during batch cultures of R. capsulatus. The experimental data were obtained by investigating the effect of different light intensities (6000-50,000 lux) on hydrogen-producing cultures of R. capsulatus grown in a batch photobioreactor, using lactate as carbon and hydrogen source. The rate of hydrogen production increased with increasing light intensity in a manner that was described by a static Baly model, modified to include the square of the light intensity. In agreement with previous studies, the kinetics of substrate utilization and growth of R. capsulatus was represented by the classical Monod or Michaelis-Menten model. When combined with a dynamic Leudekong-Piret model, the amount of hydrogen produced as a function of time was effectively predicted. These results will be useful for the automatization and control of bioprocesses for the photoproduction of hydrogen. (author)

  2. Growth kinetics of an indigenous mixed microbial consortium during phenol degradation in a batch reactor. (United States)

    Saravanan, Pichiah; Pakshirajan, K; Saha, Prabirkumar


    Biodegradation of phenol by a mixed microbial culture, isolated from a sewage treatment plant, was investigated in batch shake flasks. A minimum concentration of 100 and a maximum of 800 mg 1(-1) of phenol in the media were adapted in the degradation study. The phenol degradation rate varied largely and was less than 10 mg l(-1)h(-1) at both extremes of the initial concentrations in the media. The degradation rate was maximum 15.7 mg l(-1)h(-1) at 400 mg l(-1) phenol. The culture followed substrate inhibition kinetics and the specific growth rate were fitted to Haldane and Han-Levenspiel models. Between the two models the Han-Levenspiel was found to be a better fit with a root mean square error of 0.0211. The biokinetics constants estimated using these models showed good potential of the mixed microbial culture in phenol degradation.

  3. Fed-batch CHO cell culture for lab-scale antibody production

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Ley, Daniel; Andersen, Mikael Rørdam


    Fed-batch culture is the most commonly used upstream process in industry today for recombinant monoclonal antibody production using Chinese hamster ovary cells. Developing and optimizing this process in the lab is crucial for establishing process knowledge, which enable rapid and predictable tech......-transfer to manufacturing scale. In this chapter, we will describe stepwise how to carry out fed-batch CHO cell culture for lab-scale antibody production....

  4. Batch and High Cell Density Fed-Batch Culture Productions of an Organophosphorus Hydrolase (United States)


    0.02 g H3BO3, 0.01 g NaMoO4@ 2H2O , and 0.01 g CuSO4 . Fed-Batch Fermentations were carried out in the same Bio-Flow 3000 unit fitted with 10 L...per L): 3.0 g nitrilotriacetic acid, 6.0 MgSO4@7H2O, 1.0 g NaCl, 1.0 g MnSO4@H2O, 0.5 g FeSO4@7H20, 0.1 CaCl2@ 2H2O , 0.1 CoCl2@6H2O, 0.1 g ZnSO4@7H2O

  5. Production of Human Epidermal Growth Factor in Fed-batch Culture of Acetate-tolerant Escherichia coli%补料分批培养耐乙酸大肠杆菌生产人表皮生长因子

    Institute of Scientific and Technical Information of China (English)

    韩云; 李志敏; 杜鹏; 甘人宝; 叶勤


    An acetate-tolerant mutant of Escherichia coli DH5α, DA19, was used for secretory production of human epidermal growth factor (hEGF) whose expression was under the control of phoA promoter. The recombinant cells were cultured in a chemically defined medium, and glucose was added at different specific provision rates during the growth and expression phases. It was found that pH had a significant effect on the extracellular hEGF production. The extracellular hEGF concentration was 75.5mg·L-1, 5.2-fold of the level reached at pH 7.0, even though more acetate was produced. Nitrogen source was limited in the later growth phase. Supplementation of ammonium promoted the consumption of phosphate and reduced the time to exhaust phosphate, but the extracellular hEGF production was similar to that without supplementation of ammonium.

  6. Mixotrophic growth of Phaeodactylum tricornutum on fructose and glycerol in fed-batch and semi-continuous modes. (United States)

    Cerón-García, M C; Fernández-Sevilla, J M; Sánchez-Mirón, A; García-Camacho, F; Contreras-Gómez, A; Molina-Grima, E


    Mixotrophic cultures of Phaeodactylum tricornutum were carried out in bubble columns using fructose and glycerol in indoor fed-batch and semi-continuous modes. In the fed-batch cultures, different nutrient-addition strategies, combined with stepwise increments in the light intensity, were assayed. It was found that glycerol promoted significantly higher biomass productivity than fructose. A glycerol-induced photoinhibition that arrested the growth of P. tricornutun was also observed. As this was considered a limitation as regards transferring the fed-batch mode to outdoor conditions, this information was used to culture P. tricornutum in semi-continuous mode. Similar glycerol-induced photoinhibition was not observed in these cultures, even at highest dilution rates. Although the highest biomass (1.5 g L(-1) d(-1)) and EPA (40 mg L(-1) d(-1)) productivities found in the semi-continuous cultures were lower than those obtained photoautotrophically in outdoor photobioreactors, the findings showed that semi-continuous mode was an excellent candidate for transferring mixotrophic culture to an outdoor setting.

  7. Amino acid and glucose metabolism in fed-batch CHO cell culture affects antibody production and glycosylation

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian


    and feeds were studied using two IgG-producing cell lines. Our results indicate that the balance of glucose and amino acid concentration in the culture is important for cell growth, IgG titer and N-glycosylation. Accordingly, the ideal fate of glucose and amino acids in the culture could be mainly towards...... optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylationrelated product quality. In this work, different fed-batch processes with two chemically defined proprietary media...

  8. Decolouration of industrial metal-complex dyes in successive batches by active cultures of Trametes pubescens

    Directory of Open Access Journals (Sweden)

    Susana Rodríguez-Couto


    Full Text Available The decolouration of the metal-complex dyes Bemaplex Navy M-T (150 mg/L and Bezaktiv Blue BA (150 mg/L in nine successive batches by immobilised cultures of the white-rot fungus Trametes pubescens was studied. Two different types of immobilisation supports were used: the commercial carriers Kaldnes™ K1 (synthetic supports and sunflower-seed shells (SS (natural supports. Bemaplex showed more resistance to degradation by T. pubescens cultures than Bezaktiv, especially in the K1 cultures. Thus, SS cultures led to decolouration percentages higher than 59% for Bemaplex in all the batches save for the last two and higher than 50% for Bezaktiv in all the batches except for the 2nd and 9th ones. K1 cultures showed decolouration percentages for Bemaplex higher than 42% in batches 1, 3, 4, 5 and 7 and for Bezaktiv higher than 70% in all the batches save for the last one. Dye decolouration was mainly due to enzyme action (biodegradation.


    Directory of Open Access Journals (Sweden)

    A. C. L. Horta


    Full Text Available Abstract The performance of an in-situ capacitance sensor for on-line monitoring of biomass concentration was evaluated for some of the most important microorganisms in the biotechnology industry: Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Bacillus megaterium. A total of 33 batch and fed-batch cultures were carried out in a bench-scale bioreactor and biomass formation trends were followed by dielectric measurements during the growth phase as well as the induction phase, for 5 recombinant E. coli strains. Permittivity measurements and viable cellular concentrations presented a linear correlation for all the studied conditions. In addition, the permittivity signal was further used for inference of the cellular growth rate. The estimated specific growth rates mirrored the main trends of the metabolic states of the different cells and they can be further used for setting-up control strategies in fed-batch cultures.

  10. Bacteria obtained from a sequencing batch reactor that are capable of growth on dehydroabietic acid. (United States)

    Mohn, W W


    Eleven isolates capable of growth on the resin acid dehydroabietic acid (DhA) were obtained from a sequencing batch reactor designed to treat a high-strength process stream from a paper mill. The isolates belonged to two groups, represented by strains DhA-33 and DhA-35, which were characterized. In the bioreactor, bacteria like DhA-35 were more abundant than those like DhA-33. The population in the bioreactor of organisms capable of growth on DhA was estimated to be 1.1 x 10(6) propagules per ml, based on a most-probable-number determination. Analysis of small-subunit rRNA partial sequences indicated that DhA-33 was most closely related to Sphingomonas yanoikuyae (Sab = 0.875) and that DhA-35 was most closely related to Zoogloea ramigera (Sab = 0.849). Both isolates additionally grew on other abietanes, i.e., abietic and palustric acids, but not on the pimaranes, pimaric and isopimaric acids. For DhA-33 and DhA-35 with DhA as the sole organic substrate, doubling times were 2.7 and 2.2 h, respectively, and growth yields were 0.30 and 0.25 g of protein per g of DhA, respectively. Glucose as a cosubstrate stimulated growth of DhA-33 on DhA and stimulated DhA degradation by the culture. Pyruvate as a cosubstrate did not stimulate growth of DhA-35 on DhA and reduced the specific rate of DhA degradation of the culture. DhA induced DhA and abietic acid degradation activities in both strains, and these activities were heat labile. Cell suspensions of both strains consumed DhA at a rate of 6 mumol mg of protein-1 h-1.(ABSTRACT TRUNCATED AT 250 WORDS)

  11. Ethanol production by anaerobic thermophilic bacteria: kinetics in fed-batch cultures of Clostridium thermohydrosulfuricum

    Energy Technology Data Exchange (ETDEWEB)

    Toukourou, F.; Donaduzzi, L.; Miclo, A.; Germain, P. (Lab. of Industrial Microbiology, ENSAIA-INPL, Vandoeuvre les Nancy (FR))


    Fed-batch fermentations of Clostridium thermohydrosulfuricum are carried out using a medium rich in nitrogen source and with glucose as growth limiting factor. The ethanol/lactate yield increases as the specific growth rate and specific rate of consumption of glucose diminish. Under the experimental conditions chosen here this yield attained 3.66 moles. mole/sup -1/ with a maximal ethanol concentration of 12 g.1/sup -1/. In batch fermentation, the maximum concentration of ethanol did not exceed 8 g.1/sup -1/ independent of the concentration in glucose or nitrogen source applied. (author).

  12. Effects of plants and essential oils on ruminal in vitro batch culture methane production and fermentation (United States)

    In this study, plants (14) and essential oils (EO; 88) from plants that are naturalized to, or can be successfully grown in North America were evaluated in a batch culture in vitro screening experiments with ruminal fluid as potential anti-methanogenic additives for ruminant diets. Essential oils we...

  13. Modelling and pathway identification involving the transport mechanism of a complex metabolic system in batch culture (United States)

    Yuan, Jinlong; Zhang, Xu; Zhu, Xi; Feng, Enmin; Yin, Hongchao; Xiu, Zhilong


    The bio-dissimilation of glycerol to 1,3-propanediol (1,3-PD) by Klebsiella pneumoniae (K. pneumoniae) can be characterized by a complex metabolic system of interactions among biochemical fluxes, metabolic compounds, key enzymes and genetic regulation. In this paper, in consideration of the fact that the transport ways of 1,3-PD and glycerol with different weights across cell membrane are still unclear in batch culture, we consider 121 possible metabolic pathways and establish a novel mathematical model which is represented by a complex metabolic system. Taking into account the difficulty in accurately measuring the concentration of intracellular substances and the absence of equilibrium point for the metabolic system of batch culture, the novel approach used here is to define quantitatively biological robustness of the intracellular substance concentrations for the overall process of batch culture. To determine the most possible metabolic pathway, we take the defined biological robustness as cost function and establish an identification model, in which 1452 system parameters and 484 pathway parameters are involved. Simultaneously, the identification model is subject to the metabolic system, continuous state constraints and parameter constraints. As such, solving the identification model by a serial program is a very complicated task. We propose a parallel migration particle swarm optimization algorithm (MPSO) capable of solving the identification model in conjunction with the constraint transcription and smoothing approximation techniques. Numerical results show that the most possible metabolic pathway and the corresponding metabolic system can reasonably describe the process of batch culture.

  14. Influence of carbon source on nitrate removal by nitrate-tolerant Klebsiella oxytoca CECT 4460 in batch and chemostat cultures

    Energy Technology Data Exchange (ETDEWEB)

    Pinar, G.; Ramos, J.L. [Consejo Superior de Investigaciones Cientificas, Granada (Spain); Kovarova, K.; Egli, T. [Swiss Federal Inst. for Environmental Science and Technology, Duebendorf (Switzerland). Dept. of Microbiology


    The nitrate-tolerant organism Klebsiella oxytoca CECT-4460 tolerates nitrate at concentrations up to 1 M and is used to treat wastewater with high nitrate loads in industrial wastewater treatment plants. The authors studied the influence of the C source (glycerol or sucrose or both) on the growth rate and the efficiency of nitrate removal under laboratory conditions. With sucrose as the sole C source the maximum specific growth rate was 0.3 h{sup {minus}1}, whereas with glycerol it was 0.45 h{sup {minus}1}. In batch cultures K. oxytoca cells grown on sucrose or glycerol were able to immediately use sucrose as a sole C source, suggesting that sucrose uptake and metabolism were constitutive. In contrast, glycerol uptake occurred preferentially in glycerol-grown cells. Independent of the preculture conditions, when sucrose and glycerol were added simultaneously to batch cultures, the sucrose was used first, and once the supply of sucrose was exhausted, the glycerol was consumed. Utilization of nitrate as an N source occurred without nitrite of ammonium accumulation when glycerol was used, but nitrite accumulated when sucrose was used. In chemostat cultures K. oxytoca CECT 4460 efficiently removed nitrate without accumulation of nitrite or ammonium when sucrose, glycerol, or mixtures of these two C sources were used. The growth yields and the efficiencies of C and N utilization were determined at different growth rates in chemostat cultures. Regardless of the C source, yield carbon (Y{sub C}) ranged between 1.3 and 1.0 g (dry weight) per g of sucrose C or glycerol C consumed. Regardless of the specific growth rate and the C source, yield nitrogen (Y{sub N}) ranged from 17.2 to 12.5 g (dry weight) per g of nitrate N consumed.

  15. Changes of in vivo fluxes through central metabolic pathways during the production of nystatin by Streptomyces noursei in batch culture

    DEFF Research Database (Denmark)

    Jonsbu, E.; Christensen, Bjarke; Nielsen, Jens


    of the amino acids and calculated fluxes of the central metabolism showed that changes in the primary and secondary metabolisms occurred simultaneously. Changes in the profiles for the integrated fluxes showed a decreased flux through the pentose phosphate pathway and an increased flux in the tricarboxylic...... acid cycle relative to the glucose uptake rate when the culture entered a phase with reduced specific growth rate and enhanced nystatin yield. The flux through the pentose phosphate pathway seemed to be adjusted according to the NADPH requirement during the different phases of the batch fermentation....

  16. Acetate and ethanol production from H2 and CO2 by Moorella sp. using a repeated batch culture. (United States)

    Sakai, Shinsuke; Nakashimada, Yutaka; Inokuma, Kentaro; Kita, Masayuki; Okada, Hideki; Nishio, Naomichi


    The growth inhibition of Moorella sp. HUC22-1 by undissociated acetic acid was analyzed using a non-competitive inhibition model coupled with a pH inhibition model. In the cells grown on H2 and CO2, the inhibition constant, K(p) of the undissociated acetic acid was 6.2 mM (164 mM as the total acetate at pH 6.2, pKa = 4.795, 55 degrees C), which was 1.5-fold higher than that obtained in cells grown on fructose. When a pH-controlled batch culture was performed using a fermentor at pH 6.2 with H2 and CO2, a maximum of 0.92 g/l of dry cell weight and 339 mM of acetate were produced after 220 h, which were 4.4- and 6.8-fold higher than those produced in the pH-uncontrolled batch culture, respectively. In order to reduce acetate inhibition in the culture medium, a repeated batch culture with cell recycling was performed at a constant pH with H2 and CO2. At a pH of 6.2, the total acetate production reached 840 mmol/l-reactor with 4.7 mmol/l-reactor of total ethanol production after 420 h. When the culture pH was maintained at 5.8, which was the optimum for ethanol production, the total ethanol production reached 15.4 mmol/l-reactor after 430 h, although the total acetate production was decreased to 675 mmol/l-reactor.

  17. Batch and continuous culture kinetics for production of carotenoids by beta-ionone-resistant mutant of Xanthophyllomyces dendrorhous. (United States)

    Park, Ki-Moon; Song, Min-Woo; Kang, Seog-Jin; Lee, Jae-Heung


    A beta-ionone-resistant mutant strain isolated from the red yeast Xanthophyllomyces dendrorhous KCTC 7704 was used for batch and continuous fermentation kinetic studies with glucose media in a 2.5-1 jar fermentor at 22 degrees C and pH 4.5. The kinetic pattern of growth and carotenoid concentration in the batch fermentations exhibited a so-called mixed-growth-associated product formation, possibly due to the fact that the content of intracellular carotenoids depends on the degree of physical maturation toward adulthood. To determine the maximum specific growth rate constant (microm) and Monod constant (k(s)) for the mutant, glucose-limited continuous culture studies were performed at different dilution rates within a range of 0.02-0.10 h(-1). A reciprocal plot of the steady-state data (viz., reciprocal of glucose concentration versus residence time) obtained from continuous culture experiments was used to estimate a microm of 0.15 h(-1) and k(s) of 1.19 g/l. The carotenoid content related to the residence time appeared to assume a typical form of saturation kinetics. The maximum carotenoid content (Xm) for the mutant was estimated to be 1.04 microg/mg dry cell weight, and the Lee constant (k(m)), which was tentatively defined in this work, was found to be 3.0 h.

  18. Growth and β-galactosidase activity in cultures of Kluyveromyces marxianus under increased air pressure



    Aims: To investigate the effect of total air pressure raise on cell growth and intracellular β-galactosidase activity in batch cultures of Kluyveromyces marxianus CBS 7894. Methods and Results: A pressurized bioreactor was used for K. marxianus batch cultivation under increased air pressure from 1.2 to 6 bar. Under these conditions no inhibition of cell growth was observed. Moreover, the improvement of the oxygen transfer rate (OTR) from the gas to the culture medium by pressuriza...

  19. A comparative study on growth performance and biochemical composition of mixed culture of Isochrysis galbana and Chaetoceros calcitrans with monocultures

    Digital Repository Service at National Institute of Oceanography (India)

    Phatarpekar, P.V.; Sreepada, R.A.; Pednekar, C.; Achuthankutty, C.T.

    The growth performance, biochemical composition and nutritive value of the mixed culture of Isochrysis galbana and Chaetoceros calcitrans, grown in batch cultures under laboratory conditions was compared with those in monoculture conditions...

  20. Screening of static culture and comparison of batch and continuous culture for the textile dye biological decolorization by Phanerochaete chrysosporium

    Directory of Open Access Journals (Sweden)

    J. Urra


    Full Text Available The production of manganese dependent peroxidase (MnP by Phanerochaete chrysosporium and the level of decolorization of 13 dyes were evaluated using static and agitated batch cultures and continuous cultures. A screening carried out under static conditions showed that the oxidative system has a certain affinity for azoic structures. For concentrations of 100 mg l-1 of Acid Black 1, Reactive Black 5, Reactive Orange 16 and Acid Red 27, decolorization percentages higher than 90% were obtained. In batch cultures with Acid Black 1 and Reactive Black 5 a significant increment in primary post-metabolism biomass was observed. For these last two dyes, it was possible to explore the response of the continuous system during 32 to 47 days, with concentrations between 25 to 400 mg l-1, obtaining decolorization percentages greater than 70% for 400 mg l-1.

  1. Batch and continuous culture-based selection strategies for acetic acid tolerance in xylose-fermenting Saccharomyces cerevisiae. (United States)

    Wright, Jeremiah; Bellissimi, Eleonora; de Hulster, Erik; Wagner, Andreas; Pronk, Jack T; van Maris, Antonius J A


    Acetic acid tolerance of Saccharomyces cerevisiae is crucial for the production of bioethanol and other bulk chemicals from lignocellulosic plant-biomass hydrolysates, especially at a low pH. This study explores two evolutionary engineering strategies for the improvement of acetic acid tolerance of the xylose-fermenting S. cerevisiae RWB218, whose anaerobic growth on xylose at pH 4 is inhibited at acetic acid concentrations >1 g L(-1) : (1) sequential anaerobic, batch cultivation (pH 4) at increasing acetic acid concentrations and (2) prolonged anaerobic continuous cultivation without pH control, in which acidification by ammonium assimilation generates selective pressure for acetic acid tolerance. After c. 400 generations, the sequential-batch and continuous selection cultures grew on xylose at pH≤4 with 6 and 5 g L(-1) acetic acid, respectively. In the continuous cultures, the specific xylose-consumption rate had increased by 75% to 1.7 g xylose g(-1) biomass h(-1) . After storage of samples from both selection experiments at -80 °C and cultivation without acetic acid, they failed to grow on xylose at pH 4 in the presence of 5 g L(-1) acetic acid. Characterization in chemostat cultures with linear acetic acid gradients demonstrated an acetate-inducible acetic acid tolerance in samples from the continuous selection protocol.

  2. Monitoring and robust adaptive control of fed-batch cultures of microorganisms exhibiting overflow metabolism [abstract

    Directory of Open Access Journals (Sweden)

    Vande Wouwer, A.


    Full Text Available Overflow metabolism characterizes cells strains that are likely to produce inhibiting by-products resulting from an excess of substrate feeding and a saturated respiratory capacity. The critical substrate level separating the two different metabolic pathways is generally not well defined. Monitoring of this kind of cultures, going from model identification to state estimation, is first discussed. Then, a review of control techniques which all aim at maximizing the cell productivity of fed-batch fermentations is presented. Two main adaptive control strategies, one using an estimation of the critical substrate level as set-point and another regulating the by-product concentration, are proposed. Finally, experimental investigations of an adaptive RST control scheme using the observer polynomial for the regulation of the ethanol concentration in Saccharomyces cerevisiae fed-batch cultures ranging from laboratory to industrial scales, are also presented.

  3. Fed-batch versus batch cultures of Yarrowia lipolytica for γ-decalactone production from methyl ricinoleate. (United States)

    Gomes, Nelma; Teixeira, José A; Belo, Isabel


    Constant medium feeding rate and intermittent fed-batch fermentation strategies were investigated aiming to increase the yields of γ-decalactone production by Yarrowia lipolytica, using methyl ricinoleate as substrate and ricinoleic acid source. The accumulation of another compound, 3-hydroxy-γ-decalactone, was also analyzed since it derives from the direct precursor of γ-decalactone thereby providing information about the enzymatic activities of the pathway. Both strategies were compared with the traditional batch mode in terms of overall productivity and yield in respect to the substrate. Although the productivity of γ-decalactone was considerably higher in the batch mode (168 mg l(-1) h(-1)), substrate conversion to lactone (73 mg γ-decalactone g(-1)) was greater in the intermittent fed-batch giving 6.8 g γ-decalactone l(-1). This last strategy therefore has potential for γ-decalactone production at an industrial level.

  4. Glucose metabolism in batch and continuous cultures of Gluconacetobacter diazotrophicus PAL 3. (United States)

    Luna, María F; Bernardelli, Cecilia E; Galar, María L; Boiardi, José L


    Periplasmic glucose oxidation (by way of a pyrrolo-quinoline-quinone [PQQ]-linked glucose dehydrogenase [GDH]) was observed in continuous cultures of Gluconacetobacter diazotrophicus regardless of the carbon source (glucose or gluconate) and the nitrogen source (N(2) or NH(3)). Its synthesis was stimulated by conditions of high energetic demand (i.e., N(2)-fixation) and/or C-limitation. Under C-excess conditions, PQQ-GDH synthesis increased with the glucose concentration in the culture medium. In batch cultures, PQQ-GDH was actively expressed in very early stages with higher activities under conditions of N(2)-fixation. Hexokinase activity was almost absent under any culture condition. Cytoplasmic nicotinamide adenine dinucleotide (NAD)-linked glucose dehydrogenase (GDH) was expressed in continuous cultures under all tested conditions, and its synthesis increased with the glucose concentration. In contrast, low activities of this enzyme were detected in batch cultures. Periplasmic oxidation, by way of PQQ-GDH, seems to be the principal pathway for metabolism of glucose in G. Diazotrophicus, and NAD-GDH is an alternative route under certain environmental conditions.

  5. Multi-parameter flow cytometry and cell sorting reveal extensive physiological heterogeneity in Bacillus cereus batch cultures. (United States)

    Want, Andrew; Hancocks, Helen; Thomas, Colin R; Stocks, Stuart M; Nebe-von-Caron, Gerhard; Hewitt, Christopher J


    Based on two staining protocols, DiOC(6)(3)/propidium iodide (PI) and RedoxSensor Green (an indicator of bacterial reductase activity)/PI, multi-parameter flow cytometry and cell sorting has identified at least four distinguishable physiological states during batch cultures of Bacillus cereus. Furthermore, dependent on the position in the growth curve, single cells gave rise to varying numbers of colonies when sorted individually onto nutrient agar plates. These growing colonies derived from a single cell had widely different lag phases, inferred from differences in colony size. This further highlights the complex population dynamics of bacterial monocultures and further demonstrates that individual bacterial cells in a culture respond in markedly dissimilar ways to the environment, resulting in a physiologically heterogenous and dynamic population.

  6. Batch Growth of Chlorella Vulgaris CCALA 896 versus Semi-Continuous Regimen for Enhancing Oil-Rich Biomass Productivity

    Directory of Open Access Journals (Sweden)

    Sigita Vaičiulytė


    Full Text Available The aim of this study was to induce lipid accumulation in Chlorella cells by creating stressful growth conditions. Chlorella vulgaris CCALA 896 was grown under various batch growth modes in basal and modified BG-11 and Kolkwitz culture broths, using a continuous light regimen of 150 µE/m2/s, at 30 °C. In order to perform the experiments, two indoor photobioreactor shapes were used: a cylindrical glass photobioreactor (CGPBR with a working volume of 350 mL, and a flat glass photobioreactor (FGPBR with a working volume of 550 mL. Stress-eliciting conditions, such as nitrogen and phosphorous starvation, were imposed in order to induce lipid accumulation. The results demonstrated that more than 56% of the lipids can be accumulated in Chlorella biomass grown under two-phase batch growth conditions. The highest biomass productivity of 0.30 g/L/d was obtained at the highest nominal dilution rate (0.167 day−1 during a semi-continuous regimen, using a modified Kolkwitz medium. During the pH-stress cycles, the amount of lipids did not increase significantly and a flocculation of Chlorella cells was noted.

  7. An evaluation of the accumulation of intracellular inorganic nitrogen pools by marine microalgae in batch cultures

    Directory of Open Access Journals (Sweden)

    Paris L. Lavín


    Full Text Available Methods of extraction, changes in concentrations with growth, and effects of culture conditions on intracellular inorganic nitrogen pools (IIN - ammonia, nitrite, and nitrate were studied in nine species of marine microalgae in batch cultures. The microalgae were analysed to compare three methods of extraction of IIN, one of them developed in this study. The extraction of IIN occurs efficient by with all three methods for four out of the nine species tested. However, for five species significant differences were found among the methods, the best results being obtained with the new method. Microalgae accumulate inorganic forms of nitrogen in different proportions. The species show higher concentrations of either ammonia or nitrate, and always lower concentrations of nitrite. Microalgae of smaller cellular volumes tend to attain higher values of IIN per cubic micrometer (the converse in large-volume species, with some exceptions (Amphidinium carterae and Nannochloropsis oculata. The use of aeration in the cultures determines a decrease in the concentrations of IIN, favours nitrogen assimilation, and generates an increase in growth rates and C:N ratio. High concentrations of IIN are characteristic of the exponential growth phase, but in some cases their occurrence may result from carbon deficiency.Métodos de extração, mudanças na concentração durante o crescimento e efeitos de condições de cultivo sobre conteúdos de nitrogênio inorgânico intracelular (NII - amônia, nitrito e nitrato foram estudados em nove espécies de microalgas marinhas em cultivos estanques. As microalgas foram analisadas para comparar três métodos de extração de NII, um dos quais desenvolvido neste estudo. A extração de NII ocorre de forma eficiente com os três métodos para quatro espécies. Contudo, para cinco espécies diferenças significativas foram encontradas e os melhores resultados foram obtidos com o método novo. As microalgas acumulam formas inorg

  8. A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture. (United States)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian; Lund, Anne Mathilde; Sen, Jette Wagtberg; Rasmussen, Søren Kofoed; Kontoravdi, Cleo; Baycin-Hizal, Deniz; Betenbaugh, Michael J; Weilguny, Dietmar; Andersen, Mikael Rørdam


    In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity and quality. Titer and N-glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N-glycosylation of mAbs was dependent on the degree of starvation during early stationary phase of the fed-batch culture. Higher degree of glucose starvation reduced intracellular concentrations of UDP-GlcNAc and UDP-GalNAc, but increased the levels of UDP-Glc and UDP-Gal. Increased GlcNAc and Gal occupancy correlated well with increased degree of glucose starvation, which can be attributed to the interplay between the dilution effect associated with change in specific productivity of mAbs and the changed nucleotide sugar metabolism. Herein, we also show and discuss that increased cell culture duration negatively affect the maturation of glycans. In addition, comparative proteomics analysis of cells was conducted to observe differences in protein abundance between early growth and early stationary phases. Generally higher expression of proteins involved in regulating cellular metabolism, extracellular matrix, apoptosis, protein secretion and glycosylation was found in early stationary phase. These analyses offered a systematic view of the intrinsic properties of these cells and allowed us to explore the root causes correlating culture duration with variations in the productivity and glycosylation quality of monoclonal antibodies produced with CHO cells.

  9. Cultural diversity and economic growth

    DEFF Research Database (Denmark)

    Ager, Philipp; Brückner, Markus


    We exploit the large inflow of immigrants to the US during the 1870–1920 period to examine the effects that within-county changes in the cultural composition of the US population had on output growth. We construct measures of fractionalization and polarization to distinguish between the different...

  10. Intracellular storage of acetate/starch mixture by fast growing microbial culture in sequencing batch reactor under continuous feeding. (United States)

    Ciggin, Asli Seyhan; Majone, Mauro; Orhon, Derin


    The paper evaluated intracellular storage formation in fast growing microbial culture fed with acetate/starch mixture under continuous feeding. Three parallel laboratory-scale sequencing batch reactors (SBRs) were operated at a sludge age of 2 days: one of the SBRs was fed with acetate/starch mixture and the other two with acetate and starch, respectively, for comparing the results with single substrate systems. Despite continuous feeding, both acetate and starch components in the substrate mixture were partially converted to storage biopolymers. Poly-hydroxybutyrate (PHB) and glycogen pools were formed during SBR operation at steady state. Only a limited fraction of 12% of the acetate fed during each cycle generated PHB storage while the rest was directly utilized for microbial growth. Around half of the starch fraction of the substrate mixture was converted to glycogen. Increasing the sludge age to 8 days did not affect storage stoichiometry both for acetate and starch in the mixture.

  11. Cold-walled UHV/CVD batch reactor for the growth of Si1_x/Gex layers

    DEFF Research Database (Denmark)

    Thomsen, Erik Vilain; Christensen, Carsten; Andersen, C.R.;


    A novel cold-walled, lamp-heated, ultrahigh vacuum chemical vapor deposition (UHV/CVD) batch system for the growth of SiGe layers is presented. This system combines the batch capability of the standard UHV/CVD furnace with the temperature processing available in rapid thermal processing (Rm) equi...

  12. Acid protease and formation of multiple forms of glucoamylase in batch and continuous cultures of Aspergillus niger

    DEFF Research Database (Denmark)

    Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo;


    In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced with mole......In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced...... with molecular weights of approx. 91 (GAI), 73 (GAII), and 59 kDa (GAIII). Data from batch fermentations with constant pH 3.0 and 5.0 showed a uniform distribution of extracellular GA forms throughout the fermentations and independent of culture growth phases. Furthermore, steady-state data from chemostat...... cultivations at constant pH 3.0 and 5.0 showed a similar distribution of extracellular GA forms and established that the nitrogen concentration of the medium (C/N ratio) did not affect the distribution of multiple forms of GA. The extracellular acid protease activity was only moderate when the fungus...

  13. Photoperiodic growth enhancement in a tropical batch spawning atherinopsid, pike silverside Chirostoma estor. (United States)

    Martínez-Chávez, C C; Tello-Ballinas, A; Fonseca-Madrigal, J; Ross, L G; Martínez-Palacios, C A


    The effect of photoperiod on growth and survival in early life was determined in the tropical batch spawning atherinopsid, pike silverside Chirostoma estor. The results demonstrate high sensitivity of newly hatched C. estor to photoperiod treatments up to 90 days post hatch shown by improved growth in mass (43%) under continuous illumination. This is accompanied by increased fat deposition, which suggests a critical interaction between different photoperiod-mediated mechanisms. A thorough understanding of these mechanisms can help to optimize the development of aquaculture of C. estor and similar species.

  14. Concentrated fed-batch cell culture increases manufacturing capacity without additional volumetric capacity. (United States)

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


    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.

  15. Inferring mixed-culture growth from total biomass data in a wavelet approach

    CERN Document Server

    Ibarra-Junquera, V; Murguia-Ibarra, J S; Rosu, H C


    It is shown that the presence of mixed-culture growth in batch fermentation processes can be very accurately inferred from total biomass data by means of the wavelet analysis for singularity detection. This is accomplished by considering simple phenomenological models for the mixed growth. The main quantity provided by the wavelet analysis is the Holder exponent of the singularity that we determine for our illustrative examples. The numerical results point to the possibility that Holder exponents can be employed to characterize the nature of the mixed-culture growth in batch fermentation processes with potential industrial applications

  16. Hydrogen generation through indirect biophotolysis in batch cultures of the nonheterocystous nitrogen-fixing cyanobacterium Plectonema boryanum. (United States)

    Huesemann, Michael H; Hausmann, Tom S; Carter, Blaine M; Gerschler, Jared J; Benemann, John R


    The nitrogen-fixing nonheterocystous cyanobacterium Plectonema boryanum was used as a model organism to study hydrogen generation by indirect biophotolysis in nitrogen-limited batch cultures that were continuously illuminated and sparged with argon/CO(2) to maintain anaerobiosis. The highest hydrogen-production rate (i.e., 0.18 mL/mg day or 7.3 micromol/mg day) was observed in cultures with an initial medium nitrate concentration of 1 mM at a light intensity of 100 micromol/m(2) s. The addition of photosystem II (PSII) inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) did not reduce hydrogen-production rates relative to unchallenged controls for 50 to 150 h, and intracellular glycogen concentrations decreased significantly during the hydrogen generation period. The insensitivity of the hydrogen-production process to DCMU is indicative of the fact that hydrogen was not derived from water splitting at PSII (i.e., direct biophotolysis) but rather from electrons provided by intracellular glycogen reserves (i.e., indirect biophotolysis). It was shown that hydrogen generation could be sustained for long time periods by subjecting the cultures to alternating cycles of aerobic, nitrogen-limited growth and anaerobic hydrogen production.

  17. O2 evolution and cyclic electron flow around photosystem I in long-term ground batch culture of Euglena gracilis (United States)

    An, Yanjun; Wang, Suqin; Hao, Zongjie; Zhou, Yiyong; Liu, Yongding


    Based on the purpose of better exploring the function of green producers in the closed aquatic biological life support system, the condition of dynamic O2 evolution and performance of cyclic electron flow around photosystem I (CEF-PSI) in long-term ground batch culture of Euglena gracilis were studied, the relationship between linear electron flow (LEF) and CEF-PSI was revealed, the function of CEF-PSI was investigated. Excellent consistency in O2 evolution pattern was observed in cultures grown in both closed and open containers, O2 evolution was strictly suppressed in phase 1, but the rate of it increased significantly in phase 2. CEF-PSI was proposed to be active during the whole course of cultivation, even in the declining phase 3, it still operated at the extent of 47-55%. It is suggested that the relationship between LEF and CEF-PSI is not only competition but also reciprocity. CEF-PSI was proposed to contribute to the considerable growth in phase 1; it was also suggested to play an important protective role against photosystem II (PSII) photoinhibition at the greatly enhanced level (approximately 80-95%) on the 2nd day. Our results in this research suggest that E. gracilis had very particular photosynthetic characteristics, the strict O2 evolution suppression in the initial culture phase might be a special light acclimation behavior, and CEF-PSI could be an important mechanism involved in this kind of adaptation to the changeable light environment.

  18. Kinetics of High Cell Density Fed-batch Culture of Recombinant Escherichia coli Producing Human-like Collagen%重组大肠杆菌分批-补料高密度发酵生产类人胶原蛋白的动力学

    Institute of Scientific and Technical Information of China (English)

    花秀夫; 范代娣; 骆艳娥; 张兮; 施惠娟; 米钰; 马晓轩; 尚龙安; 赵桂仿


    The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and growth rates were set at (0.15, 0.2, 0.25h-1) by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase spectively. The model predictions are in good agreement with the experimental data.

  19. Does the 14C method estimate net photosynthesis? Implications from batch and continuous culture studies of marine phytoplankton (United States)

    Pei, Shaofeng; Laws, Edward A.


    We carried out batch culture studies with seven species of marine phytoplankton and chemostat studies with two of the seven species to determine whether and to what extent 14C uptake approximated net photosynthesis. In two of seven cases, Isochrysis galbana and Dunaliella tertiolecta, cells uniformly labeled with 14C lost no activity when they were transferred to a 14C-free medium and allowed to grow in the light. In similar experiments with four other species, uniformly labeled cells lost activity when incubated in the light, but the loss rates were only a few percent per day. Thus these six species appear to respire primarily recently fixed carbon. In the case of the remaining species, Chlorella kessleri, loss rates of 14C in the light from uniformly labeled cells were about 29% per day, the apparent ratio of respiration to net photosynthesis being 0.4. Follow-up chemostat studies with I. galbana and C. kessleri grown under both light- and nitrate-limited conditions produced results consistent with the implications of the batch culture work: uptake of 14C by I. galbana after incubations of 24 h yielded estimates of photosynthetic carbon fixation equal to the product of the chemostat dilution rate and the concentration of organic carbon in the growth chamber. Similar experiments with C. kessleri produced 14C-based estimates of photosynthetic carbon fixation that exceeded the net rates of organic carbon production in the growth chamber by roughly 55%. Time-course studies with both species indicated that at high growth rates recently fixed carbon began to enter the respiratory substrate pool after a time lag of several hours, a result consistent with previous work with D. tertiolecta. The lag time appeared to be much shorter at low growth rates. The results with C. kessleri are similar to results previously reported for Chlorella pyrenoidosa and Amphidium carteri. Collectively these results suggest that 14C uptake by species with relatively high ratios of

  20. Comparison of biomass estimation techniques for a Bacillus thuringiensis fed-batch culture

    Energy Technology Data Exchange (ETDEWEB)

    Cunha, C.C.F. [University of Newcastle upon Tyne (United Kingdom). Dept. of Chemical and Process Engineering]. E-mail:; Souza Junior, M.B. [Universidade Federal, Rio de Janeiro, RJ (Brazil). Escola de Quimica]. E-mail:


    In this work, the ability of artificial neural nets was investigated for the on-line biomass prediction of the simulated growth of a strain of Bacillus thuringiensis in fed-batch mode. For this purpose, multilayered backpropagation nets with sigmoid nodes were trained. The patterns were composed of input data on current values of biomass concentration, limiting substrate concentration and dilution rate, and output data on prediction of biomass concentration for the following step. The dilution rate was disturbed by a PRBS input, and simulations were conducted using a phenomenological experimentally validated model. The nets were able to predict the biomass concentration for different feeding techniques, and they were also compared with the variable estimation technique using the extended Kalman filter. (author)

  1. Study of Growth Mechanism of Lysozyme Crystal by Batch Crystallization Method

    Institute of Scientific and Technical Information of China (English)

    Hai Liang CUI; Yong YU; Wan Chun CHEN; Qi KANG


    The lysozyme crystals were made by batch crystallization method and the distribution of aggregate in solution were measured by dynamic light scattering. The results showed that the dimension of aggregate increased with the increase of the concentration of lysozyme and NaC1,lysozyme molecules aggregated gradually in solution and finally arrived at balance each other.The higher the concentrations of lysozyme and NaC1 were, the faster the growth rate of (110) face was. The growth rates of lysozyme crystal were obtained by a Zeiss microscope, and the effective surface energy (α) of growing steps were calculated about 4.01×l0-8 according to the model of multiple two-dimensional nucleation mechanism.

  2. Optimization of culture conditions for biological hydrogen production by Citrobacter freundii CWBI952 in batch, sequenced-batch and semicontinuous operating mode

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton, Christopher; Hiligsmann, Serge; Beckers, Laurent; Masset, Julien; Thonart, Philippe [Walloon Centre of Industrial Biology, Bd du Rectorat, 29, B.40 - P.70, B-4000 Liege (Belgium); Wilmotte, Annick [Center for Protein Engineering, Institute of Chemistry, B.6-P.14, B-4000 Liege (Belgium)


    Investigations were carried out to determine the effect of the pH, the nitrogen source, iron and the dilution rate (h{sup -1}) on fermentative hydrogen production from glucose by the newly isolated strain Citrobacter freundii CWBI952. The hydrogen production rate (HPR), hydrogen yield, biomass and soluble metabolites were monitored at 30 C in 100 mL serum bottles and in a 2.3 L bioreactor operated in batch, sequenced-batch and semicontinuous mode. The results indicate that hydrogen production activity, formate biosynthesis and glucose intake rates are very sensitive to the culture pH, and that additional formate bioconversion and production of hydrogen with lower biomass yields can be obtained at pH 5.9. In a further series of cultures casein peptone was replaced by (NH{sub 4}){sub 2}SO{sub 4}, a low cost alternative nitrogen source. The ammonia-based substitute was found to be suitable for H{sub 2} production when a concentration of 0.045 g/L FeSO{sub 4} was provided. Optimal overall performances (ca. an HPR of 33.2 mL H{sub 2}/L h and a yield of 0.83mol{sub H{sub 2}}/mol{sub glucose}) were obtained in the semicontinuous culture applying the previously optimized parameters for pH, nitrogen, and iron with a dilution rate of 0.012 h{sup -1} and degassing of biogas by N{sub 2} at a 28 mL/min flow rate. (author)

  3. Growth parameters of microalgae tolerant to high levels of carbon dioxide in batch and continuous-flow photobioreactors. (United States)

    Westerhoff, Paul; Hu, Qiang; Esparza-Soto, Mario; Vermaas, Wim


    Microalgae can be cultured in photobioreactors to sequester carbon dioxide and produce potentially valuable biomaterials. The goal of the present study was to identify and utilize microalgal strains that are capable of tolerating up to 20% CO2 (gas phase) concentrations under variable light or flue-gas blend conditions and reactor configurations to produce biomass. Scenedesmus sp. and Chlorella sp., both cultured from a Sonoran desert mineral spring, grew well and tolerated exposure to a gas mixture containing up to 20% CO2 applied continuously in batch reactors to the culture. Experiments were conducted with simulated coal-powered acidic flue gases containing SOx/NOx at concentrations of 200 to 350 ppmV. Microalgae did not grow well without pH control, and high levels (>250 mM) of nitrite or sulphite in the liquid media inhibited algal growth. Pseudo steady-state experiments were also conducted using helical tubular and flat-plate photobioreactors with continuous flow (water and gas) and with artificial or natural sunlight. With a 2 d hydraulic residence time (HRT), the helical tubular photobioreactor produced 0.50 +/- 0.11 g C d(-1) (0.056 +/- 0.012 g C L(-1) d(-1)) dry-weight cell mass during continuous fluorescent-lamp irradiance and 0.048 +/- 0.018 g C L(-1) d(-1) during 12 h light/darkness cycling. The flat-plate photobioreactor (2 d HRT) produced 0.42 +/- 0.28 g C L(-1) d(-1) with artificial lighting and with natural sunlight; a 4 d HRT produced 0.14 +/- 0.02 g C L(-1) d(-1). Reactor modelling indicated that a threshold of reactor size (i.e., HRT) an d reactor depth (path-length of light) exists based upon the optical density of the cells in the water column and their growth rates.

  4. Fermentative hydrogen yields from different sugars by batch cultures of metabolically engineered Escherichia coli DJT135

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, Dipankar; Hallenbeck, Patrick C. [Departement de Microbiologie et Immunologie, Universite de Montreal, CP 6128 succursale Centre-ville, Montreal, Quebec H3C 3J7 (Canada)


    Future sustainable production of biofuels will depend upon the ability to use complex substrates present in biomass if the use of simple sugars derived from food crops is to be avoided. Therefore, organisms capable of using a variety of fermentable carbon sources must be found or developed for processes that could produce hydrogen via fermentation. Here we have examined the ability of a metabolically engineered strain of Escherichia coli, DJT135, to produce hydrogen from glucose as well as various other carbon sources, including pentoses. The effects of pH, temperature and carbon source were investigated in batch experiments. Maximal hydrogen production from glucose was obtained at an initial pH of 6.5 and temperature of 35 C. Kinetic growth studies showed that the {mu}max was 0.0495 h{sup -1} with a Ks of 0.0274 g L{sup -1} when glucose was the sole carbon source in M9 (1X) minimal medium. Among the many sugar and sugar derivatives tested, hydrogen yields were highest with fructose, sorbitol and D-glucose; 1.27, 1.46 and 1.51 mol H{sub 2} mol{sup -1} substrate respectively. (author)

  5. Fed-batch culture of Escherichia coli for L-valine production based on in silico flux response analysis. (United States)

    Park, Jin Hwan; Kim, Tae Yong; Lee, Kwang Ho; Lee, Sang Yup


    We have previously reported the development of a 100% genetically defined engineered Escherichia coli strain capable of producing L-valine from glucose with a high yield of 0.38 g L-valine per gram glucose (0.58 mol L-valine per mol glucose) by batch culture. Here we report a systems biological strategy of employing flux response analysis in bioprocess development using L-valine production by fed-batch culture as an example. Through the systems-level analysis, the source of ATP was found to be important for efficient L-valine production. There existed a trade-off between L-valine production and biomass formation, which was optimized for the most efficient L-valine production. Furthermore, acetic acid feeding strategy was optimized based on flux response analysis. The final fed-batch cultivation strategy allowed production of 32.3 g/L L-valine, the highest concentration reported for E. coli. This approach of employing systems-level analysis of metabolic fluxes in developing fed-batch cultivation strategy would also be applicable in developing strategies for the efficient production of other bioproducts.


    NARCIS (Netherlands)

    Klasing, Mariko J.; Milionis, Petros


    To what extent does the cultural composition of a society impose a constraint on its long-run growth potential? We study this question in the context of an innovation-based model of growth where cultural attitudes are endogenously transmitted from one generation to the next. Focusing on attitudes re

  7. Magnesium Uptake by the Green Microalga Chlorella vulgaris in Batch Cultures. (United States)

    Ben Amor-Ben Ayed, Hela; Taidi, Behnam; Ayadi, Habib; Pareau, Dominique; Stambouli, Moncef


    The accumulation (internal and superficial distribution) of magnesium ions (Mg(2+)) by the green freshwater microalga Chlorella vulgaris (C. vulgaris) was investigated under autotrophic culture in a stirred photobioreactor. The concentrations of the three forms of Mg(2+) (dissolved, extracellular, and intracellular) were determined with atomic absorption spectroscopy during the course of C. vulgaris growth. The proportions of adsorbed (extracellular) and absorbed (intracellular) Mg(2+) were quantified. The concentration of the most important pigment in algal cells, chlorophyll a, increased over time in proportion to the increase in the biomass concentration, indicating a constant chlorophyll/biomass ratio during the linear growth phase. The mean-average rate of Mg(2+) uptake by C. vulgaris grown in a culture medium starting with 16 mg/l of Mg(2+) concentration was measured. A clear relationship between the biomass concentration and the proportion of the Mg(2+) removal from the medium was observed. Of the total Mg(2+) present in the culture medium, 18% was adsorbed on the cell wall and 51% was absorbed by the biomass by the end of the experiment (765 h). Overall, 69% of the initial Mg(2+) were found to be removed from the medium. This study supported the kinetic model based on a reversible first-order reaction for Mg(2+) bioaccumulation in C. vulgaris, which was consistent with the experimental data.

  8. Effects of CO2 enrichment and nutrients supply intermittency on batch cultures of Isochrysis galbana. (United States)

    Picardo, Marta C; de Medeiros, José Luiz; Araújo, Ofélia de Queiroz F; Chaloub, Ricardo Moreira


    Aiming at enhanced performance to increase economic feasibility of microalgae based processes, Isochrysis galbana was grown in three modes of cultivation: batch, intermittent fed batch and semi-continuous. The batch mode was conducted under two regimes of aeration: conventional aeration and CO2 enriched aeration (5% v/v in air). Increased biomass productivity without significant impact on lipid accumulation was observed for CO2 enriched aeration relatively to cultivation aerated with air only. The intermittent fed batch cultivation policy was proven to be useful for lipid accumulation, increasing the lipid content by 19.8%. However, the semi-continuous mode resulted in higher productivity due to increased biomass concentration; the biomass productivity reached 0.51 g/(Ld). Fluorescence measurements were performed; the calculated low electron transport rate showed the need to increase the irradiance. The results showed that I. galbana can be grown in semi-continuous condition at high levels of biomass productivity.

  9. A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian;


    In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity and qual......In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity...... and quality. Titer and N-glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N-glycosylation of mAbs was dependent on the degree of starvation during early stationary...... phase of the fed-batch culture. Higher degree of glucose starvation reduced intracellular concentrations of UDP-GlcNAc and UDP-GalNAc, but increased the levels of UDP-Glc and UDP-Gal. Increased GlcNAc and Gal occupancy correlated well with increased degree of glucose starvation, which can be attributed...

  10. Batch By Batch Longitudinal Emittance Blowup MD

    CERN Document Server

    Mastoridis, T; Butterworth, A; Jaussi, M; Molendijk, J


    The transverse bunch emittance increases significantly at 450 GeV from the time of injection till the ramp due to IBS. By selectively blowing up the longitudinal emittance of the incoming batch at each injection, it should be possible to reduce the transverse emittance growth rates due to IBS. An MD was conducted on April 22nd 2012 to test the feasibility and performance of the batch-by-batch longitudinal emittance blowup. There were three main goals during the MD. First, to test the developed hardware, firmware, and software for the batch-by-batch blowup. Then, to measure the transverse emittance growth rates of blown-up and "witness" batches to quantify any improvement, and finally to test the ALLInjectSequencer class, which deals with the complicated gymnastics of introducing or masking the new batch to various RF loops.

  11. Differential effect of culture temperature and specific growth rate on CHO cell behavior in chemostat culture. (United States)

    Vergara, Mauricio; Becerra, Silvana; Berrios, Julio; Osses, Nelson; Reyes, Juan; Rodríguez-Moyá, María; Gonzalez, Ramon; Altamirano, Claudia


    Mild hypothermia condition in mammalian cell culture technology has been one of the main focuses of research for the development of breeding strategies to maximize productivity of these production systems. Despite the large number of studies that show positive effects of mild hypothermia on specific productivity of r-proteins, no experimental approach has addressed the indirect effect of lower temperatures on specific cell growth rate, nor how this condition possibly affects less specific productivity of r-proteins. To separately analyze the effects of mild hypothermia and specific growth rate on CHO cell metabolism and recombinant human tissue plasminogen activator productivity as a model system, high dilution rate (0.017 h(-1)) and low dilution rate (0.012 h(-1)) at two cultivation temperatures (37 and 33 °C) were evaluated using chemostat culture. The results showed a positive effect on the specific productivity of r-protein with decreasing specific growth rate at 33 °C. Differential effect was achieved by mild hypothermia on the specific productivity of r-protein, contrary to the evidence reported in batch culture. Interestingly, reduction of metabolism could not be associated with a decrease in culture temperature, but rather with a decrease in specific growth rate.

  12. Brunovsky Normal Form of Monod Kinetics Models and Growth Rate Control of a Fed-batch Cultivation Process

    Directory of Open Access Journals (Sweden)

    Pavlov Y.


    Full Text Available A mathematical methodology that gives assistance to design of fed-batch stabilization and control is presented. The methodology is based both on Utility theory and optimal Control theory. The Utility theory deals with the expressed subjective preferences and allows for the expert preferences to be taken in consideration in complex biotechnological systems as criteria for control and optimization. The Control theory is used for parameters stabilization of a fed-batch cultivation process. The control is written based on information of the growth rate. The simulations show good efficiency of the control laws.

  13. Bacteriocin production by strain Lactobacillus delbrueckii ssp. bulgaricus BB18 during continuous prefermentation of yogurt starter culture and subsequent batch coagulation of milk. (United States)

    Simova, E D; Beshkova, D M; Angelov, M P; Dimitrov, Zh P


    By screening for bacteriocin-producing lactic acid bacteria of 1,428 strains isolated from authentic Bulgarian dairy products, Lb. bulgaricus BB18 strain obtained from kefir grain was selected. Out of 11 yogurt starters containing Lb. bulgaricus BB18 and S. thermophilus strains resistant to bacteriocin secreted by Lb. bulgaricus BB18 a yogurt culture (S. thermophilus 11A+Lb. bulgaricus BB18) with high growth and bacteriocinogenic activity in milk was selected. Continuous (pH-stat 5.7) prefermentation processes were carried out in milk at 37 degrees C in a 2l MBR bioreactor (MBR AG, Zurich, Switzerland) with an IMCS controller for agitation speed, temperature, dissolved oxygen, CO2 and pH. Prefermented milk with pH 5.7 coagulated in a thermostat at 37 degrees C until pH 4.8-4.9. S. thermophilus 11A and Lb. bulgaricus BB18 grew independently in a continuous mode at similar and sufficiently high-dilution rates (D=1.83 h(-1)-S. thermophilus 11A; D=1.80 h(-1)-Lb. bulgaricus BB18). The yogurt cultures developed in a stream at a high-dilution rate (D=2.03-2.28 h(-1)). The progress of both processes (growth and bacteriocin production) depended on the initial ratio between the two microorganisms. The continuous prefermentation process promoted conditions for efficient fermentation and bacteriocinogenesis of the starter culture during the batch process: strong reduction of the times for bacteriocin production and coagulation of milk (to 4.5-5.0 h); high cell productivity (lactobacilli-4x10(12) CFU ml(-1), streptococci-6x10(12) CFU ml(-1)); high productivity of bacteriocins (4,500 BU ml(-1))-1.7 times higher than the bacteriocinogenic activity of the batch starter culture.

  14. Monitoring and Characterization of Crystal Nucleation and Growth during Batch Crystallization

    NARCIS (Netherlands)

    Kadam, S.S.


    Batch crystallization is commonly used in pharmaceutical, agrochemical, specialty and fine chemicals industry. The advantages of batch crystallization lie in its ease of operation and the relatively simple equipment that can be used. On the other hand a major disadvantage associated with it is the i

  15. Novel micro-bioreactor high throughput technology for cell culture process development: Reproducibility and scalability assessment of fed-batch CHO cultures. (United States)

    Amanullah, Ashraf; Otero, Jose Manuel; Mikola, Mark; Hsu, Amy; Zhang, Jinyou; Aunins, John; Schreyer, H Brett; Hope, James A; Russo, A Peter


    With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell platform operated under fed-batch conditions could support viable cell concentrations up to least 12 x 10(6) cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) x 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within +/-20% of the historical averages.

  16. Acid protease and formation of multiple forms of glycoamylase in batch and continuous cultures of Aspergillus niger

    DEFF Research Database (Denmark)

    Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo


    In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced...... degradation of the GA forms at low pH. It was concluded that the observed modifications of the extracellular profile of GA isoforms in A. niger Bo-1 are due to changes in pH and medium composition....

  17. Growth of the coccolithophore Emiliania huxleyi in light- and nutrient-limited batch reactors: relevance for the BIOSOPE deep ecological niche of coccolithophores (United States)

    Perrin, Laura; Probert, Ian; Langer, Gerald; Aloisi, Giovanni


    Coccolithophores are unicellular calcifying marine algae that play an important role in the oceanic carbon cycle via their cellular processes of photosynthesis (a CO2 sink) and calcification (a CO2 source). In contrast to the well-studied, surface-water coccolithophore blooms visible from satellites, the lower photic zone is a poorly known but potentially important ecological niche for coccolithophores in terms of primary production and carbon export to the deep ocean. In this study, the physiological responses of an Emiliania huxleyi strain to conditions simulating the deep niche in the oligotrophic gyres along the BIOSOPE transect in the South Pacific Gyre were investigated. We carried out batch culture experiments with an E. huxleyi strain isolated from the BIOSOPE transect, reproducing the in situ conditions of light and nutrient (nitrate and phosphate) limitation. By simulating coccolithophore growth using an internal stores (Droop) model, we were able to constrain fundamental physiological parameters for this E. huxleyi strain. We show that simple batch experiments, in conjunction with physiological modelling, can provide reliable estimates of fundamental physiological parameters for E. huxleyi that are usually obtained experimentally in more time-consuming and costly chemostat experiments. The combination of culture experiments, physiological modelling and in situ data from the BIOSOPE cruise show that E. huxleyi growth in the deep BIOSOPE niche is limited by availability of light and nitrate. This study contributes more widely to the understanding of E. huxleyi physiology and behaviour in a low-light and oligotrophic environment of the ocean.

  18. Isomaltulose production using free cells: optimisation of a culture medium containing agricultural wastes and conversion in repeated-batch processes. (United States)

    Kawaguti, Haroldo Y; Buzzato, Michele F; Sato, Hélia H


    The enzyme glucosyltransferase is an industrially important enzyme since it produces non-cariogenic isomaltulose (6-O-alpha-D-glucopyronosyl-1-6-D-fructofuranose) from sucrose by intramolecular transglucosylation. The experimental designs and response surface methodology (RSM) were applied for the optimisation of the nutrient concentrations in the culture medium for the production of glucosyltransferase by Erwinia sp. D12 in shaken flasks at 200 rpm and 30 degrees C. A statistical analysis of the results showed that, in the range studied, the factors had a significant effect (P free Erwinia sp. D12 cells in a batch process using an orbital shaker. The influence of the parameters sucrose concentration, temperature, pH, and cell concentration on the conversion of sucrose into isomaltulose was studied. The free cells showed a high conversion rate of sucrose into isomaltulose using batch fermentation, obtaining an isomaltulose yield of 72.11% from sucrose solution 35% at 35 degrees C.

  19. Autotrophic growth and lipid production of Chlorella sorokiniana in lab batch and BIOCOIL photobioreactors: Experiments and modeling. (United States)

    Concas, Alessandro; Malavasi, Veronica; Costelli, Cristina; Fadda, Paolo; Pisu, Massimo; Cao, Giacomo


    A novel mathematical model for the quantitative assessment of the effect of dissolved nitrogen on the autotrophic batch-growth and lipid accumulation of Chlorella sorokiniana, is proposed in this work. Model results have been validated through comparison with suitable experimental data performed in lab photobioreactors. Further experiments have been then performed using the BIOCOIL photobioreactor operated in fed-batch mode. The experimental results, which show that a maximum growth rate of 0.52day(-1) and a lipid content equal to 25%wt can be achieved with the BIOICOIL, have been successfully predicted through the proposed model. Therefore, the model might represent a first step toward the development of a tool for the scale-up and optimization of the operating conditions of BIOCOIL photobioreactors. Finally, the fatty acid methyl esters obtained by trans-esterification of lipids extracted from C. sorokiniana, have been analyzed in view of the assessment of their usability for producing biodiesel.

  20. Optimisation of batch culture conditions for cyclodextrin glucanotransferase production from Bacillus circulans DF 9R

    Directory of Open Access Journals (Sweden)

    Krymkiewicz Norberto


    Full Text Available Abstract Background The extracellular enzyme cyclodextrin glucanotransferase (CGTase synthesizes cyclic malto-oligosaccharides called cyclodextrins (CDs from starch and related α-1,4-glucans. CGTases are produced by a variety of bacteria, mainly Bacillus species, by submerged culture in complex medium. CGTases differ in the amount and types of CDs produced. In addition, CGTase production is highly dependent on the strain, medium composition and culture conditions. Therefore we undertook this study with a newly isolated strain of Bacillus circulans. Results CGTase activity produced from Bacillus circulans DF 9R was optimised in shake flasks using a combination of conventional sequential techniques and statistical experimental design. Effects of nutrients, including several carbon, nitrogen and mineral sources, were assayed. The selected minimal medium consisted of 1.5 % cassava starch, 0.4 % ammonium sulphate, 0.1 M phosphate buffer, 0.002 % MgSO4 and 0.002 % FeSO4. The optimal concentrations of carbon and nitrogen sources were determined using a central composite design. Maximum CGTase activity obtained in supernatants was 5.8 U/mL in 48 h of incubation. Optimal conditions for enzyme production also included an initial pH of 8.3 and 37°C as the incubation temperature. Cell growth and CGTase production profile were not linked to each other, suggesting that enzyme production/secretion is not growth–associated but mainly a late-log phase event. Conclusion We have screened conditions for optimal CGTase production. The selected minimal medium contained starch, ammonium, Mg2+ and Fe2+ as essential nutrients. As an additional advantage, this medium does not require complex nitrogen sources with varying and unknown composition.

  1. Microbial succession in response to pollutants in batch-enrichment culture


    Shuo Jiao; Weimin Chen; Entao Wang; Junman Wang; Zhenshan Liu; Yining Li; Gehong Wei


    As a global problem, environmental pollution is an important factor to shape the microbial communities. The elucidation of the succession of microbial communities in response to pollutants is essential for developing bioremediation procedures. In the present study, ten batches of soil-enrichment subcultures were subjected to four treatments: phenanthrene, n-octadecane, phenanthrene + n-octadecane, or phenanthrene + n-octadecane + CdCl2. Forty pollutant-degrading consortia, corresponding to ea...

  2. Novel approach of high cell density recombinant bioprocess development: Optimisation and scale-up from microlitre to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures

    Directory of Open Access Journals (Sweden)

    Rimšeliene Renata


    Full Text Available Abstract Background Bioprocess development of recombinant proteins is time consuming and laborious as many factors influence the accumulation of the product in the soluble and active form. Currently, in most cases the developmental line is characterised by a screening stage which is performed under batch conditions followed by the development of the fed-batch process. Performing the screening already under fed-batch conditions would limit the amount of work and guarantee that the selected favoured conditions also work in the production scale. Results Here, for the first time, high throughput multifactorial screening of a cloning library is combined with the fed-batch technique in 96-well plates, and a strategy is directly derived for scaling to bioreactor scale. At the example of a difficult to express protein, an RNase inhibitor, it is demonstrated that screening of various vector constructs and growth conditions can be performed in a coherent line by (i applying a vector library with promoters and ribosome binding sites of different strength and various fusion partners together with (ii an early stage use of the fed-batch technology. It is shown that the EnBase® technology provides an easy solution for controlled cultivation conditions in the microwell scale. Additionally the high cell densities obtained provide material for various analyses from the small culture volumes. Crucial factors for a high yield of the target protein in the actual case were (i the fusion partner, (ii the use of of a mineral salt medium together with the fed-batch technique, and (iii the preinduction growth rate. Finally, it is shown that the favorable conditions selected in the microwell plate and shake flask scales also work in the bioreactor. Conclusions Cultivation media and culture conditions have a major impact on the success of a screening procedure. Therefore the application of controlled cultivation conditions is pivotal. The consequent use of fed-batch

  3. A novel fed-batch based cultivation method provides high cell-density and improves yield of soluble recombinant proteins in shaken cultures

    Directory of Open Access Journals (Sweden)

    Glumoff Tuomo


    Full Text Available Abstract Background Cultivations for recombinant protein production in shake flasks should provide high cell densities, high protein productivity per cell and good protein quality. The methods described in laboratory handbooks often fail to reach these goals due to oxygen depletion, lack of pH control and the necessity to use low induction cell densities. In this article we describe the impact of a novel enzymatically controlled fed-batch cultivation technology on recombinant protein production in Escherichia coli in simple shaken cultures. Results The enzymatic glucose release system together with a well-balanced combination of mineral salts and complex medium additives provided high cell densities, high protein yields and a considerably improved proportion of soluble proteins in harvested cells. The cultivation method consists of three steps: 1 controlled growth by glucose-limited fed-batch to OD600 ~10, 2 addition of growth boosters together with an inducer providing efficient protein synthesis within a 3 to 6 hours period, and 3 a slow growth period (16 to 21 hours during which the recombinant protein is slowly synthesized and folded. Cell densities corresponding to 10 to 15 g l-1 cell dry weight could be achieved with the developed technique. In comparison to standard cultures in LB, Terrific Broth and mineral salt medium, we typically achieved over 10-fold higher volumetric yields of soluble recombinant proteins. Conclusions We have demonstrated that by applying the novel EnBase® Flo cultivation system in shaken cultures high cell densities can be obtained without impairing the productivity per cell. Especially the yield of soluble (correctly folded proteins was significantly improved in comparison to commonly used LB, Terrific Broth or mineral salt media. This improvement is thought to result from a well controlled physiological state during the whole process. The higher volumetric yields enable the use of lower culture volumes and can

  4. Sporulation of Clostridium cellulolyticum while grown in cellulose-batch and cellulose-fed continuous cultures on a mineral-salt based medium. (United States)

    Desvaux, M; Petitdemange, H


    Clostridium cellulolyticum sporulation was investigated during growth on cellulose fibers in a mineral-salt based medium which corresponds to conditions linked to its natural ecological niche. At steady state of the continuous cultures under limitation and with an excess of cellulose and/or ammonium, bacterial cells mainly sporulated at low dilution rates (D), at least 10% sporulation being observed at the lowest D tested. Increasing the cellulose concentration in the feed-medium reservoir increased the percentage of spores in the bioreactor. It appeared that the remaining undigested cellulose could serve as an exogenous carbon source supply at a continuous but limited rate throughout the sporulation process. In addition to the proportion of carbon and nitrogen, the influence of the environmental pH on spore formation was studied. In cellulose-fed continuous cultures at a constant D and a pH decreasing from 7.2 to 6.4, the percentage of spores increased to 14% at the lowest pH tested. When C. cellulolyticum was grown in batch culture, the level of sporulation was dramatically higher in unregulated-pH fermentation compared to pH-controlled growth conditions at pH 7.2 since in the former it reached 45% within 5 days of cultivation. It then appeared that a low specific growth rate and a low environmental pH in the presence of an insoluble carbon substrate were the major factors inducing sporulation in C. cellulolyticum. Furthermore, since the spores adhere to the carbon substrate (the cellulose) the bacteria gain advantages when the environment allows germination thanks to the recovery of suitable growth conditions. By allowing the maintenance and the integrity of the bacteria in the microbiota, spore formation could then explain the successful survival of C. cellulolyticum in cellulosic anaerobic habitats where low environmental pH conditions are often found.

  5. Expression and purification of recombinant human granulocyte colony-stimulating factor in fed-batch culture of Escherichia coli. (United States)

    Kim, Chang-Kyu; Choi, Jun-Ha; Lee, Seung-Bae; Lee, Sang-Mahn; Oh, Jae-Wook


    Granulocyte colony-stimulating factor (G-CSF) is a cytokine that has multiple roles in hematopoietic cells such as the regulation of proliferation and differentiation. Here, we describe fed-batch culture, refolding, and purification of rhG-CSF. The suitability of urea or sarcosine for solubilizing inclusion bodies (IBs) was tested. It was observed that urea is more efficient for solubilizing and refolding IBs than sarcosine is. The purity of rhG-CSF and the removal percentage of the rhG-CSF isoforms during purification were increased by pH 5.5 precipitation. The purity and the yield of purified rhG-CSF were 99% and 0.5 g of protein per liter culture broth, respectively. Our protocols of recombinant protein purification using ion exchange chromatography and semipreparative high performance liquid chromatography of pH-precipitated refolded solution may be informative to the industrial scale production of biopharmaceuticals.

  6. Bench to batch: advances in plant cell culture for producing useful products. (United States)

    Weathers, Pamela J; Towler, Melissa J; Xu, Jianfeng


    Despite significant efforts over nearly 30 years, only a few products produced by in vitro plant cultures have been commercialized. Some new advances in culture methods and metabolic biochemistry have improved the useful potential of plant cell cultures. This review will provide references to recent relevant reviews along with a critical analysis of the latest improvements in plant cell culture, co-cultures, and disposable reactors for production of small secondary product molecules, transgenic proteins, and other products. Some case studies for specific products or production systems are used to illustrate principles.

  7. A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian;


    In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity...... and quality. Titer and N-glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N-glycosylation of mAbs was dependent on the degree of starvation during early stationary...... phase of the fed-batch culture. Higher degree of glucose starvation reduced intracellular concentrations of UDP-GlcNAc and UDP-GalNAc, but increased the levels of UDP-Glc and UDP-Gal. Increased GlcNAc and Gal occupancy correlated well with increased degree of glucose starvation, which can be attributed...

  8. Microbial succession in response to pollutants in batch-enrichment culture. (United States)

    Jiao, Shuo; Chen, Weimin; Wang, Entao; Wang, Junman; Liu, Zhenshan; Li, Yining; Wei, Gehong


    As a global problem, environmental pollution is an important factor to shape the microbial communities. The elucidation of the succession of microbial communities in response to pollutants is essential for developing bioremediation procedures. In the present study, ten batches of soil-enrichment subcultures were subjected to four treatments: phenanthrene, n-octadecane, phenanthrene + n-octadecane, or phenanthrene + n-octadecane + CdCl2. Forty pollutant-degrading consortia, corresponding to each batch of the four treatments were obtained. High-throughput sequencing of the 16S rRNA gene revealed that the diversity, richness and evenness of the consortia decreased throughout the subculturing procedure. The well-known hydrocarbon degraders Acinetobacter, Gordonia, Sphingobium, Sphingopyxis, and Castellaniella and several other genera, including Niabella and Naxibacter, were detected in the enriched consortia. The predominant microbes varied and the microbial community in the consortia gradually changed during the successive subculturing depending on the treatment, indicating that the pollutants influenced the microbial successions. Comparison of the networks in the treatments indicated that organic pollutants and CdCl2 affected the co-occurrence patterns in enriched consortia. In conclusion, single environmental factors, such as the addition of nutrients or selection pressure, can shape microbial communities and partially explain the extensive differences in microbial community structures among diverse environments.

  9. A fully defined, fed-batch, recombinant NS0 culture process for monoclonal antibody production. (United States)

    Hermes, Paul A; Castro, Chris D


    To manufacture a glycoprotein, mammalian cells expressing the desired protein are often grown in fed-batch mode. Feeding an undefined, nonanimal hydrolysate helps the cells receive sufficient nutrition, but makes systems difficult to optimize. Even different lots of the same hydrolysate may have significant variability; furthermore, individual components may actually be detrimental to the cells. Switching to fully defined feeds could eliminate these issues. For monoclonal antibody (mAb) production by fed-batch NS0 cells, this article describes the replacement of a hydrolysate-based feed with a fully defined, animal-component-free feed system. The defined feed initially had 67 components, but additional experiments allowed a reduction to 25 components. The mAb titer is approximately 20% higher than in the undefined system, and the feed volume is circa 20% lower. The two systems generated antibodies with similar glycosylation profiles. Other benefits of the defined feed system include lower raw material costs, the ability to optimize key nutrient concentrations, greater confidence in raw material quality, and the elimination of potential, hydrolysate-associated endotoxin issues.

  10. Ammonium nitrogen removal in batch cultures treating digested piggery wastewater with microalgae Oedogonium sp. (United States)

    Wang, Haiping; Hu, Zhiquan; Xiao, Bo; Cheng, Qunpeng; Li, Fanghua


    Due to the nutrient characteristics of the high concentration of available ammonium in digested piggery wastewater (DPW), microalgae can be used to treat DPW before its final discharge. Four green microalgae (Hydrodictyaceae reticulatum Lag, Scenedesmus obliquus, Oedogonium sp. and Chlorella pyrenoidosa) and three blue-green algae (Anabaena flos-aquae, Oscillatoria amoena Gom and Spirulina platensis) were used to remove the nutrients (N, P, C), especially ammonium nitrogen (NH4(+)-N), from diluted DPW with 300 mg/L algae density in batch tests. The microalgae with the best NH4(+)-N nutrient removal was then selected for further optimization of the variables to improve NH4(+)-N removal efficiency using a central composite design (CCD) experiment. Taking into account the nutrient removal efficiency, Oedogonium sp. showed the best performance (reduction of 95.9% NH4(+)-N, 92.9% total phosphorus (TP) and 62.5% chemical oxygen demand (COD)) based on the results of the batch tests. The CCD results suggested that the optimal values of variables were initial Oedogonium sp. density of 399.2 mg/L and DPW diluted by 16.3, while the predicted value of NH4(+)-N removal efficiency obtained was 97.0%.

  11. Dynamic model-based analysis of furfural and HMF detoxification by pure and mixed batch cultures of S. cerevisiae and S. stipitis. (United States)

    Hanly, Timothy J; Henson, Michael A


    Inhibitory compounds that result from biomass hydrolysis are an obstacle to the efficient production of second-generation biofuels. Fermentative microorganisms can reduce compounds such as furfural and 5-hydroxymethyl furfural (HMF), but detoxification is accompanied by reduced growth rates and ethanol yields. In this study, we assess the effects of these furan aldehydes on pure and mixed yeast cultures consisting of a respiratory deficient mutant of Saccharomyces cerevisiae and wild-type Scheffersomyces stipitis using dynamic flux balance analysis. Uptake kinetics and stoichiometric equations for the intracellular reduction reactions associated with each inhibitor were added to genome-scale metabolic reconstructions of the two yeasts. Further modification of the S. cerevisiae metabolic network was necessary to satisfactorily predict the amount of acetate synthesized during HMF reduction. Inhibitory terms that captured the adverse effects of the furan aldehydes and their corresponding alcohols on cell growth and ethanol production were added to attain qualitative agreement with batch experiments conducted for model development and validation. When the two yeasts were co-cultured in the presence of the furan aldehydes, inoculums that reduced the synthesis of highly toxic acetate produced by S. cerevisiae yielded the highest ethanol productivities. The model described here can be used to generate optimal fermentation strategies for the simultaneous detoxification and fermentation of lignocellulosic hydrolysates by S. cerevisiae and/or S. stipitis.

  12. Optimisation of batch culture conditions for cell-envelope-associated proteinase production from Lactobacillus delbrueckii subsp. lactis ATCC® 7830™. (United States)

    Agyei, Dominic; Potumarthi, Ravichandra; Danquah, Michael K


    Using a combination of conventional sequential techniques, the batch growth conditions for the production of cell-envelope-associated proteinases have for the first time been studied and optimised in Lactobacillus delbrueckii subsp. lactis 313 (ATCC 7830; LDL 313). Concentrations of inoculum (0.1 production medium (0.2 production included an initial pH of 6.0, 45 °C incubation temperature, 2 % (v/v) inoculum size of OD(560) = 1, 150 rpm agitation speed, and growth medium carbon/nitrogen ratio of 1.0. Maximum proteinase activity obtained for whole cells was 0.99 U/ml after 8 h of incubation. The variables studied are very relevant due to their significance in improving the productivity of proteinase synthesis from LDL 313, under process and, likely, economic optimum conditions.


    COOK, T M


    Cook, Thomas M. (Rutgers, The State University, New Brunswick, N.J.). Growth of Thiobacillus thiooxidans in shaken culture. J. Bacteriol. 88:620-623. 1964.-When flasks of sulfur-containing medium were inoculated with Thiobacillus thiooxidans and placed on a shaking machine immediately after inoculation, growth was markedly inhibited. If various wetting agents were provided, or if the ratio of inoculum to sulfur was increased, growth under shaking conditions was possible. Evidence cited shows that the natural wetting agent (or other externally supplied surfactants) is necessary to effect a bacteria-sulfur contact which is stable to shaking conditions.

  14. Kinetics of sugars consumption and ethanol inhibition in carob pulp fermentation by Saccharomyces cerevisiae in batch and fed-batch cultures. (United States)

    Lima-Costa, Maria Emília; Tavares, Catarina; Raposo, Sara; Rodrigues, Brígida; Peinado, José M


    The waste materials from the carob processing industry are a potential resource for second-generation bioethanol production. These by-products are small carob kibbles with a high content of soluble sugars (45-50%). Batch and fed-batch Saccharomyces cerevisiae fermentations of high density sugar from carob pods were analyzed in terms of the kinetics of sugars consumption and ethanol inhibition. In all the batch runs, 90-95% of the total sugar was consumed and transformed into ethanol with a yield close to the theoretical maximum (0.47-0.50 g/g), and a final ethanol concentration of 100-110 g/l. In fed-batch runs, fresh carob extract was added when glucose had been consumed. This addition and the subsequent decrease of ethanol concentrations by dilution increased the final ethanol production up to 130 g/l. It seems that invertase activity and yeast tolerance to ethanol are the main factors to be controlled in carob fermentations. The efficiency of highly concentrated carob fermentation makes it a very promising process for use in a second-generation ethanol biorefinery.

  15. Simulation and prediction of protein production in fed-batch E. coli cultures: An engineering approach. (United States)

    Calleja, Daniel; Kavanagh, John; de Mas, Carles; López-Santín, Josep


    An overall model describing the dynamic behavior of fed-batch E. coli processes for protein production has been built, calibrated and validated. Using a macroscopic approach, the model consists of three interconnected blocks allowing simulation of biomass, inducer and protein concentration profiles with time. The model incorporates calculation of the extra and intracellular inducer concentration, as well as repressor-inducer dynamics leading to a successful prediction of the product concentration. The parameters of the model were estimated using experimental data of a rhamnulose-1-phosphate aldolase-producer strain, grown under a wide range of experimental conditions. After validation, the model has successfully predicted the behavior of different strains producing two different proteins: fructose-6-phosphate aldolase and ω-transaminase. In summary, the presented approach represents a powerful tool for E. coli production process simulation and control.

  16. Viral and bacterial production in the North Water: in situ measurements, batch-culture experiments and characterization and distribution of a virus host system (United States)

    Middelboe, Mathias; Nielsen, Torkel G.; Bjørnsen, Peter K.

    Growth and viral lysis of bacterioplankton at subzero temperatures were measured in the North Water polynya in July 1998. In situ measurements of bacterial carbon consumption in surface waters ranged from 15 to 63 μg C l -1 d -1 in the eastern and 6 to 7 μg C l -1 d -1 in the northern part of the polynya. Both bacterial abundance and activity appeared to increase in response to the decay of the phytoplankton bloom that developed in the North Water. Organic carbon was the limiting substrate for bacteria in the polynya since addition of glucose, but not inorganic nutrients, to batch cultures increased both the carrying capacity of the substrate and the growth rate of the bacteria. Bacterial growth rates ranged from 0.11 to 0.40 d -1, corresponding to bacterial generation times of 1.7-6.3 d. The in situ viral production rate was estimated both from the frequency of visibly infected cells and from the rate of viral production in batch cultures; it ranged from 0.04 to 0.52 d -1 and from 0.25 to 0.47 d -1, respectively. From 6% to 28% of bacterial production was found to be lost due to viral lysis. The average virus-bacteria ratio was 5.1±3.1, with the abundance of viruses being correlated positively with bacterial production. A Pseudoalteromonas sp. bacterial host and an infective virus were isolated from the polynya; characteristics and distribution of the virus-host system were examined. The Pseudoalteromonas sp. showed psychrotolerant growth and sustained significant production of viruses at 0°C. The virus-host system was found throughout the polynya. Overall the results suggested that a large amount of organic carbon released during the development and breakdown of the spring phytoplankton bloom was consumed by planktonic bacteria and that the microbial food web was an important and dynamic component of the planktonic food web in the North Water.

  17. Fingerprint detection and process prediction by multivariate analysis of fed-batch monoclonal antibody cell culture data. (United States)

    Sokolov, Michael; Soos, Miroslav; Neunstoecklin, Benjamin; Morbidelli, Massimo; Butté, Alessandro; Leardi, Riccardo; Solacroup, Thomas; Stettler, Matthieu; Broly, Hervé


    This work presents a sequential data analysis path, which was successfully applied to identify important patterns (fingerprints) in mammalian cell culture process data regarding process variables, time evolution and process response. The data set incorporates 116 fed-batch cultivation experiments for the production of a Fc-Fusion protein. Having precharacterized the evolutions of the investigated variables and manipulated parameters with univariate analysis, principal component analysis (PCA) and partial least squares regression (PLSR) are used for further investigation. The first major objective is to capture and understand the interaction structure and dynamic behavior of the process variables and the titer (process response) using different models. The second major objective is to evaluate those models regarding their capability to characterize and predict the titer production. Moreover, the effects of data unfolding, imputation of missing data, phase separation, and variable transformation on the performance of the models are evaluated.

  18. Batch fermentative hydrogen production by enriched mixed culture: Combination strategy and their microbial composition. (United States)

    Sivagurunathan, Periyasamy; Sen, Biswarup; Lin, Chiu-Yue


    The effect of individual and combined mixed culture on dark fermentative hydrogen production performance was investigated. Mixed cultures from cow dung (C1), sewage sludge (C2), and pig slurry (C3) were enriched under strict anaerobic conditions at 37°C with glucose as the sole carbon source. Biochemical hydrogen production test in peptone-yeast-glucose (PYG) and basal medium was performed for individual mixed cultures (C1, C2 and C3) and their combinations (C1-C2, C2-C3, C1-C3 and C1-C2-C3) at a glucose concentration of 10 g/L, 37°C and initial pH 7. Maximum hydrogen yields (HY) of 2.0 and 1.86 [Formula: see text] by C2, and 1.98 and 1.95 mol(H2)/mol(glucose) by C2-C3 were obtained in PYG and basal medium, respectively. Butyrate and acetate were the major soluble metabolites produced by all the cultures, and the ratio of butyrate to acetate was ∼2 fold higher in basal medium than PYG medium, indicating strong influence of media formulation on glucose catabolism. The major hydrogen-producing bacterial strains, observed in all mixed cultures, belonged to Clostridium butyricum, C. saccharobutylicum, C. tertium and C. perfringens. The hydrogen production performance of the combined mixed culture (C2-C3) was further evaluated on beverage wastewater (10 g/L) at pH 7 and 37°C. The results showed an HY of 1.92 mol(H2)/mol(glucose-equivalent). Experimental evidence suggests that hydrogen fermentation by mixed culture combination could be a novel strategy to improve the HY from industrial wastewater.

  19. Enhancement of fermentative hydrogen production in an extreme-thermophilic (70°C) mixed-culture environment by repeated batch cultivation. (United States)

    Lu, Wenjing; Fan, Gaoyuan; Zhao, Chenxi; Wang, Hongtao; Chi, Zifang


    Repeated batch cultivation was applied to enrich hydrogen fermentative microflora under extreme-thermophilic (70°C) environment. Initial inoculums received from a hydrogen producing reactor fed with organic fraction of household solid wastes. In total seven transfers was conducted and maximum hydrogen yield reached 296 ml H(2)/g (2.38 mol/mol) glucose and 252 ml H(2)/g (2.03 mol/mol) for 1 and 2 g/l glucose medium, respectively. It was found that hydrogen production was firstly decreased and got increased gradually from third generation. Acetate was found to be the main metabolic by-product in all batch cultivation. Furthermore, the diversity of bacterial community got decreased after repeated batch cultivation. It was proved that repeated batch cultivation was a good method to enhance the hydrogen production by enriching the mixed cultures of dominant species.

  20. Treatment of agro based industrial wastewater in sequencing batch reactor: performance evaluation and growth kinetics of aerobic biomass. (United States)

    Lim, J X; Vadivelu, V M


    A sequencing batch reactor (SBR) with a working volume of 8 L and an exchange ratio of 25% was used to enrich biomass for the treatment of the anaerobically treated low pH palm oil mill effluent (POME). The influent concentration was stepwise increased from 5000 ± 500 mg COD/L to 11,500 ± 500 mg COD/L. The performance of the reactor was monitored at different organic loading rates (OLRs). It was found that approximately 90% of the COD content of the POME wastewater was successfully removed regardless of the OLR applied to the SBR. Cycle studies of the SBR show that the oxygen uptake by the biomass while there is no COD reduction may be due to the oxidation of the storage product by the biomass. Further, the growth kinetic parameters of the biomass were determined in batch experiments using respirometer. The maximum specific growth rate (μmax) was estimated to be 1.143 day(-1) while the half saturation constant (Ks) with respect to COD was determined to be 0.429 g COD/L. The decay coefficient (bD) and biomass yield (Y) were found to be 0.131 day(-1) and 0.272 mg biomass/mg COD consumed, respectively.

  1. Variations in ribulose 1,5-bisphosphate carboxylase protein levels, activities and subcellular distribution during photoautotrophic batch culture of Chlorogloeopsis fritschii. (United States)

    Lanaras, T; Codd, G A


    Ribulose 1,5-bisphosphate (RuBP) carboxylase is present in the cytoplasm and carboxysomes (polyhedral bodies) of the cyanobacterium Chlorogloeopsis fritschii. In vitro enzyme activities have been measured throughout photoautotrophic batch culture, together with RuBP carboxylase protein concentrations, determined by rocket immunoelectrophoresis. Enzyme activities and protein levels in the cytoplasmic and carboxysomal fractions varied in an apparently inverse manner during growth. The RuBP carboxylase activities per unit enzyme protein were maximal in late lag phase/early exponential phase for both cellular enzyme pools. Both rates per unit enzyme protein declined during exponential phase, cytoplasmic enzyme activity remaining consistently higher than that of the carboxysomal enzyme. Activities per unit cytoplasmic and carboxysomal enzyme protein showed very low, similar rates in late stationary phase and death phase. Dialysis experiments indicated that such changes were not due to interference in activity assays by soluble endogenous effectors. Major shifts in the subcellular distribution of RuBP carboxylase protein were found versus culture age, enzyme protein levels being predominantly carboxysomal in lag phase, mainly soluble in exponential phase and then mainly carboxysomal again in stationary/death phase. The data are discussed in terms of carboxysome function and the question of control of RuBP carboxylase synthesis in cyanobacteria.

  2. Polyhydroxybutyrate production by direct use of waste activated sludge in phosphorus-limited fed-batch culture. (United States)

    Cavaillé, Laëtitia; Grousseau, Estelle; Pocquet, Mathieu; Lepeuple, Anne-Sophie; Uribelarrea, Jean-Louis; Hernandez-Raquet, Guillermina; Paul, Etienne


    Polyhydroxybutyrate (PHB) production directly by waste activated sludge (WAS) was investigated in aerobic fed-batch conditions using acetic acid as substrate. PHB production was induced by phosphorus limitation. WAS of different origin were tested with various degrees of phosphorus limitation and PHB contents of up to 70% (gCOD PHB/gCOD particulate) were obtained. This strategy showed the importance of maintaining cell growth for PHB production in order to increase PHB concentration and that the degree of phosphorus limitation has a direct impact on the quantity of PHB produced. Pyrosequencing of 16S rRNA transcripts showed changes in the active bacteria of the WAS microbial community as well as the acclimation of populations depending on sludge origin. The monitoring of the process appeared as the key factor for optimal PHB production by WAS. Different strategies are discussed and compared in terms of carbon yield and PHB content with the feast and famine selection process.

  3. Kinetic study on fermentation from CO[sub 2] and H[sub 2] using the acclimated-methanogen in batch culture

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Z.Y.; Maekawa, Takaaki (Tsukuba Univ. (Japan). Inst. of Agricultural and Forest Engineering)


    Methane was produced from H[sub 2] and CO[sub 2] using the acclimated-mixed methanogens in a 3.71 fermentor in batch culture at pH 7.2 and 37[sup o]C. The fermentation kinetics parameter for the growth of methanogens, overall mass transfer coefficient of the reactor, and the conversion rate of H[sub 2] and CO[sub 2] to CH[sub 4] by the acclimated-mixed culture were determined. The maximum specific growth rate (mu[sub max]) and H[sub 2] specific consumption rate (q[sub max]) were found to be 0.064 (h[sup -1]) and 104.8 (mmol h[sup -1] g[sup -1]), respectively. Monod saturation constants for growth (Kp) and for inhibition (K'p) were found to be 3.54 (kPa) and 0.57 (kPa), respectively. These findings indicate that without very low dissolved H[sub 2] levels, the fermentation are carried out under [mu][sub max], and the specific uptake rate (q) was almost not affected at any dissolved H[sub 2] level in the range studied. The yield of CH[sub 4] (Yp/s) was calculated to be 0.245 (mol CH[sub 4] mol[sup -1] H[sub 2]), which is near the stoichiometric value of 0.25. DH[sub 2] was also measured using the Teflon tubing method and was in good agreement with those estimated by kinetic calculations. (author)


    Energy Technology Data Exchange (ETDEWEB)

    Weldy, C.S.; Huesemann, M.


    Atmospheric carbon dioxide (CO2) concentrations are increasing and may cause unknown deleterious environmental effects if left unchecked. The Intergovernmental Panel on Climate Change (IPCC) has predicted in its latest report a 2°C to 4°C increase in global temperatures even with the strictest CO2 mitigation practices. Global warming can be attributed in large part to the burning of carbon-based fossil fuels, as the concentration of atmospheric CO2 is directly related to the burning of fossil fuels. Biofuels which do not add CO2 to the atmosphere are presently generated primarily from terrestrial plants, i.e., ethanol from corn grain and biodiesel from soybean oil. The production of biofuels from terrestrial plants is severely limited by the availability of fertile land. Lipid production from microalgae and its corresponding biodiesel production have been studied since the late 1970s but large scale production has remained economically infeasible due to the large costs of sterile growing conditions required for many algal species. This study focuses on the potential of the halophilic microalgae species Dunaliella salina as a source of lipids and subsequent biodiesel production. The lipid production rates under high light and low light as well as nitrogen suffi cient and nitrogen defi cient culture conditions were compared for D. salina cultured in replicate photobioreactors. The results show (a) cellular lipid content ranging from 16 to 44% (wt), (b) a maximum culture lipid concentration of 450mg lipid/L, and (c) a maximum integrated lipid production rate of 46mg lipid/L culture*day. The high amount of lipids produced suggests that D. salina, which can be mass-cultured in non-sterile outdoor ponds, has strong potential to be an economically valuable source for renewable oil and biodiesel production.

  5. Enhancing toxic protein expression in Escherichia coli fed-batch culture using kinetic parameters: Human granulocyte-macrophage colony-stimulating factor as a model system. (United States)

    Khasa, Yogender Pal; Khushoo, Amardeep; Mukherjee, Krishna Jyoti


    The kinetics of recombinant human granulocyte-macrophage colony-stimulating factor (hGM-CSF) expression was studied under the strong T7 promoter in continuous culture of Escherichia coli using complex medium to design an optimum feeding strategy for high cell density cultivation. Continuous culture studies were done at different dilution rates and the growth and product formation profiles were monitored post-induction. Recombinant protein expression was in the form of inclusion bodies with a maximum specific product formation rate (q(p)) of 63.5 mg g(-1) DCW h(-1) at a dilution rate (D) of 0.3 h(-1). The maximum volumetric product concentration achieved at this dilution rate was 474 mg l(-1), which translated a ~1.4 and ~1.75 folds increase than the values obtained at dilution rates of 0.2 h(-1) and 0.4 h(-1) respectively. The specific product yield (Y(P/x)) peaked at 138 mg g(-1) DCW, demonstrating a ~1.6 folds increase in the values obtained at other dilution rates. A drop in q(p) was observed within 5-6 h of induction at all the dilution rates, possibly due to protein toxicity and metabolic stress associated with protein expression. The data from the continuous culture studies allowed us to design an optimal feeding strategy and induction time in fed-batch cultures which resulted in a maximum product concentration of 3.95 g l(-1) with a specific hGM-CSF yield (Y(P/x)) of 107 mg g(-1) DCW.

  6. Variação da composição química do meio de cultura e da bioquímica celular de Phaeodactylum tricomutum (Bohlin, em cultivos estanques Long term changes in the culture medium composition and cellular biochemistry of Phacodactylum tricomutum (Bohlin, in batch cultures

    Directory of Open Access Journals (Sweden)

    Elizabeth Aidar


    Full Text Available P. tricornutum was grown in batch cultures under 21ºC temperature and continuous irradiance of 183 µE.m-2 . s-1, provided by fluorescent light tubes. Cultures growth were followed by cell countings, "in vivo" fluorescence and cellular protein, carbohydrate and chlorophyll-α content. The maximum value of protein/carbohydrate ratio was found at the exponential phase and the minimum value at the stationary phase. This ratio characterizes the physiological stage of the culture. Cell uptake rates for nitrate, ammonium and phosphate from the media were higher during the lag-phase. Nitrite excretion by the cells were detected following nitrate uptake.

  7. Impact of seed loading ratio on the growth kinetics of mono-ammonium phosphate under isothermal batch crystallization

    Energy Technology Data Exchange (ETDEWEB)

    Long, Bingwen; Yang, Haotian; Ding, Yigang [Wuhan Institute of Technology, Wuhan (China)


    The effect of seed load ratio on the growth kinetics of Mono-ammonium phosphate (MAP) under isothermal batch crystallization was investigated quantitatively. A direct parameter estimation method was proposed and applied to extract the growth kinetic parameters from a simple crystallization model using our experimental solution concentration decline data. The method assured the globally best parameters to be obtained and was found less sensitive to experimental errors. The linear growth constants k{sub g} and the growth order g were found to be in the range of 1,000-2,600 μm·min{sup -1} and 0.93-1.12, respectively, for MAP crystallized at 40 .deg. C. Both parameters decreased significantly with increase of seed load ratio and k{sub g} even showed a strong linear decline trend. The effective crystallization time also decreased with the seed mass. The proposed methodology could be extended to study the effect of other operation variables such as temperature and initial supersaturation on the crystal growth rate.

  8. Glutathione accumulation in ethanol-stat fed-batch culture of Saccharomyces cerevisiae with a switch to cysteine feeding. (United States)

    Nisamedtinov, Ildar; Kevvai, Kaspar; Orumets, Kerti; Rautio, Jari J; Paalme, Toomas


    Shot-wise supplementation of cysteine to a yeast culture is a common means of promoting glutathione (GSH) production. In the present work, we study the accumulation kinetics of cysteine, gamma-glutamylcysteine, and GSH and the expression of genes involved in GSH and sulfur metabolism in ethanol-stat fed-batch cultures as a result of switching to a medium enriched with cysteine and glycine. Supplementation in this fashion resulted in a rapid but short-term increase in the rate of GSH synthesis, while the expression of GSH1 decreased. Expression of GSH1 and GSH synthesis rate were observed to revert close to the base level after a few hours. These results indicate that, under such conditions, the control of GSH synthesis at higher concentrations occurred at the enzymatic, rather than the transcriptional level. The incorporation of cysteine into GSH was limited to approximately 40% of the theoretical yield, due to its requirement as a source of sulfur for protein synthesis under conditions whereby the sulfate assimilation pathway is down-regulated. This was supported by the expression profiles of genes involved in cysteine and homocysteine interconversion.

  9. Production of functional killer protein in batch cultures upon a shift from aerobic to anaerobic conditions

    Directory of Open Access Journals (Sweden)

    Gildo Almeida da Silva


    Full Text Available The aim of this work was to study the production of functional protein in yeast culture. The cells of Saccharomyces cerevisiae Embrapa 1B (K+R+ killed a strain of Saccharomyces cerevisiae Embrapa 26B (K-R-in grape must and YEPD media. The lethal effect of toxin-containing supernatant and the effect of aeration upon functional killer production and the correlation between the products of anaerobic metabolism and the functional toxin formation were evaluated. The results showed that at low sugar concentration, the toxin of the killer strain of Sacch. cerevisiae was only produced under anaerobic conditions . The system of killer protein production showed to be regulated by Pasteur and Crabtree effects. As soon as the ethanol was formed, the functional killer toxin was produced. The synthesis of the active killer toxin seemed to be somewhat associated with the switch to fermentation process and with concomitant alcohol dehydrogenase (ADH activity.

  10. On-line monitoring of important organoleptic methyl-branched aldehydes during batch fermentation of starter culture Staphylococcus xylosus reveal new insight into their production in a model fermentation

    DEFF Research Database (Denmark)

    de Vos Petersen, Christian; Beck, Hans Christian; Lauritsen, Frants R


    of a longer aerobic growth period. Growing S. xylosus under conditions resembling those in a fermented sausage revealed that NaCl (5%) increased aldehyde production considerably, whereas KNO(3) (0.03%) or NaNO(2) (0.03%) had little effect. A lowering of pH from 7.2 to 6.0 reduced cell density, but had a minor......A small fermentor (55 mL) was directly interfaced to a membrane inlet mass spectrometer for continuous on-line monitoring of oxygen and volatile metabolites during batch fermentations of the starter culture Staphylococcus xylosus. Using this technique, we were able to correlate production...

  11. A new bioenergetic and thermodynamic approach to batch photoautotrophic growth of Arthrospira (Spirulina) platensis in different photobioreactors and under different light conditions. (United States)

    da Silva, Milena Fernandes; Casazza, Alessandro Alberto; Ferrari, Pier Francesco; Perego, Patrizia; Bezerra, Raquel Pedrosa; Converti, Attilio; Porto, Ana Lucia Figueiredo


    Photobioreactor configuration, mode of operation and light intensity are known to strongly impact on cyanobacteria growth. To shed light on these issues, kinetic, bioenergetic and thermodynamic parameters of batch Arthrospira platensis cultures were estimated along the time at photosynthetic photon flux density (PPFD) of 70μmolm(-2)s(-1) in different photobioreactors with different surface/volume ratio (S/V), namely open pond (0.25cm(-1)), shaken flask (0.48cm(-1)), horizontal photobioreactor (HoP) (1.94cm(-1)) and helicoidal photobioreactor (HeP) (3.88cm(-1)). Maximum biomass concentration and productivity remarkably increased with S/V up to 1.94cm(-1). HoP was shown to be the best-performing system throughout the whole runs, while HeP behaved better only at the start. Runs carried out in HoP increasing PPFD from 40 to 100μmolm(-2)s(-1) revealed a progressive enhancement of bioenergetics and thermodynamics likely because of favorable light distribution. HoP appeared to be a promising configuration to perform high-yield indoor cyanobacterial cultures.

  12. Phenol biodegradation using a repeated batch culture of Candida tropicalis in a multistage bubble column. (United States)

    Ruiz-Ordaz, N; Ruiz-Lagunez, J C; Castañon-González, J H; Hernández-Manzano, E; Cristiani-Urbina, E; Galíndez-Mayer, J


    As in many other microorganisms, the growth rate of C. tropicalis is affected by phenol. Besides, when the yeast is aerobically cultivated in a medium containing phenol, using a bubble column, the yeast cell flotation phenomenon occurs, which makes the continuous operation of this type of reactor difficult. Therefore, a system of phenol degradation, which recycles the biomass separated by flotation, was devised in this work. In order to reduce the substrate toxicity observed at high phenol concentrations, the bubble column used in the biodegradation studies was fed in a semibatch mode. So, a semicontinuous system was implemented to treat effluents with relatively high concentrations (> 9,000 ppm) of phenol, by replacing periodically about 22% of the bioreactor operational volume. The phenol removal efficiencies obtained with this system were higher than 98.7%.

  13. Induction of secondary metabolism of Aspergillus terreus ATCC 20542 in the batch bioreactor cultures. (United States)

    Boruta, Tomasz; Bizukojc, Marcin


    Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification of process conditions and growth media. Several strategies were tested, including the addition of rapeseed oil or inulin, changing the concentration of nitrogen source, reduction of chlorine supply, cultivation under saline conditions, and using various aeration schemes. Fifteen secondary metabolites were identified in the course of the study by using ultra-high performance liquid chromatography coupled with mass spectrometry, namely mevinolinic acid, 4a,5-dihydromevinolinic acid, 3α-hydroxy-3,5-dihydromonacolin L acid, terrein, aspulvinone E, dihydroisoflavipucine, (+)-geodin, (+)-bisdechlorogeodin, (+)-erdin, asterric acid, butyrolactone I, desmethylsulochrin, questin, sulochrin, and demethylasterric acid. The study also presents the collection of mass spectra that can serve as a resource for future experiments. The growth in a salt-rich environment turned out to be strongly inhibitory for secondary metabolism and the formation of dense and compact pellets was observed. Generally, the addition of inulin, reducing the oxygen supply, and increasing the content of nitrogen source did not enhance the production of examined molecules. The most successful strategy involved the addition of rapeseed oil to the chlorine-deficient medium. Under these conditions, the highest levels of butyrolactone I, asterric acid, and mevinolinic acid were achieved and the presence of desmethylsulochrin and (+)-bisdechlorogeodin was detected in the broth. The constant and relatively high aeration rate in the idiophase was shown to be beneficial for terrein and (+)-geodin biosynthesis.

  14. Growth and exopolysaccharide yield of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081 in batch and continuous bioreactor experiments at constant pH. (United States)

    Mende, Susann; Krzyzanowski, Leona; Weber, Jost; Jaros, Doris; Rohm, Harald


    Some Lactobacillus delbrueckii ssp. bulgaricus strains are able to synthesize exopolysaccharides (EPS) and are therefore highly important for the dairy industry as starter cultures. The aim of this study was to investigate the nutritional requirements for growth and EPS production of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081. A medium was developed from a semi-defined medium (SDM) in which glucose was replaced by lactose and different combinations of supplements (nucleobases, vitamins, salts, sodium formate and orotic acid) were added. Constant pH batch fermentation with the modified medium resulted in an EPS yield of approximately 210 mg glucose equivalents per liter medium. This was a 10-fold increase over flask cultivation of this strain in SDM. Although not affecting cell growth, the mixture of salts enhanced the EPS synthesis. Whereas EPS production was approximately 12 mg/g dry biomass without salt supplementation, a significantly higher yield (approximately 20 mg/g dry biomass) was observed after adding the salt mixture. In continuous fermentation, a maximal EPS concentration was obtained at a dilution rate of 0.31/h (80 mg EPS/L), which corresponded to a specific EPS production of 49 mg/g dry biomass.

  15. Fractionation and characterization of polysaccharides from cyanobacterium Spirulina (Arthrospira) maxima in nitrogen-limited batch culture

    Institute of Scientific and Technical Information of China (English)


    By a sequentially selective extraction procedure, polysaccharides (PS) from cyanobacterium Spirulina (Arthrospira) maxima that can be bio-functionally healthy components were fractionated in four parts including the culture medium (PSCM), the external layers of the cell (PSEL), the cell wall (PSCW) and storage granules (PSSG). The four fractionated parts of polysaccharides were characterized by gas-liquid chromatography (GLC) equipped with FID and fused-silica capillary column (15 m×0.53 mm i. d.). The contents of PSCM, PSEL, PSCW and PSSG were about 2.0%, 2.6%, 10.0% and 52.0% to cell dry matter, respectively. Glucose was almost the only monosaccharide in PSSG and PSCW and most predominant in PSEL, while in PSCM, xylose, rhamnose and glucose were the main ones. Two uronic acids represented by glucuronic acid and galacturoic acid, six neutral monosaccharides including fucose, rhamnose, xylose, mannose, galactose and glucose, and two possible unidentified sugars were found in PSEL and PSCM. These data are valuable for the selective productions of high-added value sugars from Spirulina.


    Directory of Open Access Journals (Sweden)

    C. Cruz-Martínez


    Full Text Available AbstractNH4NO3 simultaneously provides a readily assimilable nitrogen source (ammonia and a reserve of nitrogen (nitrate, allowing for an increase in Arthrospira platensis biomass production while reducing the cost of the cultivation medium. In this study, a 22plus star central composite experimental design combined with response surface methodology was employed to analyze the influence of light intensity (I and the total amount of added NH4NO3 (Mt on a bench-scale tubular photobioreactor for fed-batch cultures. The maximum cell concentration (Xm, cell productivity (PX and biomass yield on nitrogen (YX/N were evaluated, as were the protein and lipid contents. Under optimized conditions (I = 148 μmol·photons·m-2·s-1 and Mt = 9.7 mM NH4NO3, Xm = 4710 ±34.4 mg·L-1, PX = 478.9 ±3.8 mg·L-1·d-1 and YX/N = 15.87 ±0.13 mg·mg-1 were obtained. The best conditions for protein content in the biomass (63.2% were not the same as those that maximized cell growth (I = 180 μmol·photons·m-2·s-1 and Mt = 22.5 mM NH4NO3. Based on these results, it is possible to conclude that ammonium nitrate is an interesting alternate nitrogen source for the cultivation of A. platensisin a fed-batch process and could be used for other photosynthetic microorganisms.

  17. Effect of algae growth on aerobic granulation and nutrients removal from synthetic wastewater by using sequencing batch reactors. (United States)

    Huang, Wenli; Li, Bing; Zhang, Chao; Zhang, Zhenya; Lei, Zhongfang; Lu, Baowang; Zhou, Beibei


    The effect of algae growth on aerobic granulation and nutrients removal was studied in two identical sequencing batch reactors (SBRs). Sunlight exposure promoted the growth of algae in the SBR (Rs), forming an algal-bacterial symbiosis in aerobic granules. Compared to the control SBR (Rc), Rs had a slower granulation process with granules of loose structure and smaller particle size. Moreover, the specific oxygen uptake rate was significantly decreased for the granules from Rs with secretion of 25.7% and 22.5% less proteins and polysaccharides respectively in the extracellular polymeric substances. Although little impact was observed on chemical oxygen demand (COD) removal, algal-bacterial symbiosis deteriorated N and P removals, about 40.7-45.4% of total N and 44% of total P in Rs in contrast to 52.9-58.3% of TN and 90% of TP in Rc, respectively. In addition, the growth of algae altered the microbial community in Rs, especially unfavorable for Nitrospiraceae and Nitrosomonadaceae.

  18. Modelling of Batch Process Operations

    DEFF Research Database (Denmark)


    Here a batch cooling crystalliser is modelled and simulated as is a batch distillation system. In the batch crystalliser four operational modes of the crystalliser are considered, namely: initial cooling, nucleation, crystal growth and product removal. A model generation procedure is shown...

  19. Genetic Algorithmic Optimization of PHB Production by a Mixed Culture in an Optimally Dispersed Fed-batch Bioreactor

    Directory of Open Access Journals (Sweden)

    Pratap R. Patnaik


    Full Text Available Poly-β-hydroxybutyrate (PHB is an energy-storage polymer whose properties are similar to those of chemical polymers such as polyethylene and polypropylene. Moreover, PHB is biodegradable, absorbed by human tissues and less energy-consuming than synthetic polymers. Although Ralstonia eutropha is widely used to synthesize PHB, it is inefficient in utilizing glucose and similar sugars. Therefore a co-culture of R. eutropha and Lactobacillus delbrueckii is preferred since the latter can convert glucose to lactate, which R. eutropha can metabolize easily. Tohyama et al. [24] maximized PHB production in a well-mixed fed-batch bioreactor with glucose and (NH42SO4 as the primary substrates. Since production-scale bioreactors often deviate from ideal laboratory-scale reactors, a large bioreactor was simulated by means of a dispersion model with the kinetics determined by Tohyama et al. [24] and dispersion set at an optimum Peclet number of 20 [32]. The time-dependent feed rates of the two substrates were determined through a genetic algorithm (GA to maximize PHB production. This bioreactor produced 22.2% more PHB per liter and 12.8% more cell mass than achieved by Tohyama et al. [24]. These results, and similar observations with other fermentations, indicate the feasibility of enhancing the efficiency of large nonideal bioreactors through GA optimizations.

  20. Characteristics of human cell line, F2N78, for the production of recombinant antibody in fed-batch and perfusion cultures. (United States)

    Seo, Joon Serk; Min, Byung Sub; Kwon, Young-Bum; Lee, Soo-Young; Cho, Jong-Moon; Park, Keun-Hee; Yang, Yae Ji; Maeng, Ki Eun; Chang, Shin-Jae; Kim, Dong-Il


    A human hybrid cell line, F2N78, was developed by somatic fusion of HEK293 and Namalwa cells for the production recombinant biopharmaceutical proteins. In this study, we performed perfusion culture to verify its potential in culture process used for human cell expression platform. Cell viability could be maintained over 90% and high viable cell density was obtained at higher than 1.0 × 10(7) cells/mL by bleeding process in perfusion culture. The cells were adapted well in both culture modes, but there were apparent differences in protein quality. Compared to fed-batch culture, degalactosylated forms such as G0F and G0 as well as Man5 showed no significant increases in perfusion culture. In terms of charge variants, acidic peaks increased, whereas main peaks constantly decreased according to the length of culture period in both methods.

  1. Mixed-culture transcriptome analysis reveals the molecular basis of mixed-culture growth in Streptococcus thermophilus and Lactobacillus bulgaricus. (United States)

    Sieuwerts, Sander; Molenaar, Douwe; van Hijum, Sacha A F T; Beerthuyzen, Marke; Stevens, Marc J A; Janssen, Patrick W M; Ingham, Colin J; de Bok, Frank A M; de Vos, Willem M; van Hylckama Vlieg, Johan E T


    Many food fermentations are performed using mixed cultures of lactic acid bacteria. Interactions between strains are of key importance for the performance of these fermentations. Yogurt fermentation by Streptococcus thermophilus and Lactobacillus bulgaricus (basonym, Lactobacillus delbrueckii subsp. bulgaricus) is one of the best-described mixed-culture fermentations. These species are believed to stimulate each other's growth by the exchange of metabolites such as folic acid and carbon dioxide. Recently, postgenomic studies revealed that an upregulation of biosynthesis pathways for nucleotides and sulfur-containing amino acids is part of the global physiological response to mixed-culture growth in S. thermophilus, but an in-depth molecular analysis of mixed-culture growth of both strains remains to be established. We report here the application of mixed-culture transcriptome profiling and a systematic analysis of the effect of interaction-related compounds on growth, which allowed us to unravel the molecular responses associated with batch mixed-culture growth in milk of S. thermophilus CNRZ1066 and L. bulgaricus ATCC BAA-365. The results indicate that interactions between these bacteria are primarily related to purine, amino acid, and long-chain fatty acid metabolism. The results support a model in which formic acid, folic acid, and fatty acids are provided by S. thermophilus. Proteolysis by L. bulgaricus supplies both strains with amino acids but is insufficient to meet the biosynthetic demands for sulfur and branched-chain amino acids, as becomes clear from the upregulation of genes associated with these amino acids in mixed culture. Moreover, genes involved in iron uptake in S. thermophilus are affected by mixed-culture growth, and genes coding for exopolysaccharide production were upregulated in both organisms in mixed culture compared to monocultures. The confirmation of previously identified responses in S. thermophilus using a different strain combination

  2. Metabolic Control in Mammalian Fed-Batch Cell Cultures for Reduced Lactic Acid Accumulation and Improved Process Robustness

    Directory of Open Access Journals (Sweden)

    Viktor Konakovsky


    Full Text Available Biomass and cell-specific metabolic rates usually change dynamically over time, making the “feed according to need” strategy difficult to realize in a commercial fed-batch process. We here demonstrate a novel feeding strategy which is designed to hold a particular metabolic state in a fed-batch process by adaptive feeding in real time. The feed rate is calculated with a transferable biomass model based on capacitance, which changes the nutrient flow stoichiometrically in real time. A limited glucose environment was used to confine the cell in a particular metabolic state. In order to cope with uncertainty, two strategies were tested to change the adaptive feed rate and prevent starvation while in limitation: (i inline pH and online glucose concentration measurement or (ii inline pH alone, which was shown to be sufficient for the problem statement. In this contribution, we achieved metabolic control within a defined target range. The direct benefit was two-fold: the lactic acid profile was improved and pH could be kept stable. Multivariate Data Analysis (MVDA has shown that pH influenced lactic acid production or consumption in historical data sets. We demonstrate that a low pH (around 6.8 is not required for our strategy, as glucose availability is already limiting the flux. On the contrary, we boosted glycolytic flux in glucose limitation by setting the pH to 7.4. This new approach led to a yield of lactic acid/glucose (Y L/G around zero for the whole process time and high titers in our labs. We hypothesize that a higher carbon flux, resulting from a higher pH, may lead to more cells which produce more product. The relevance of this work aims at feeding mammalian cell cultures safely in limitation with a desired metabolic flux range. This resulted in extremely stable, low glucose levels, very robust pH profiles without acid/base interventions and a metabolic state in which lactic acid was consumed instead of being produced from day 1. With

  3. Conveyance Systems and Vegetative Growth in Peach (Prunuspersica (l. Batch high low Tropical Conditions

    Directory of Open Access Journals (Sweden)

    Luis Gómez


    Full Text Available Fruit trees known as deciduous species are native to the northern temperate zones that are well adapted to environmental conditions in regions of high Colombian Andean tropics, especially in the highlands boyacense where approximately 80% of the planted area. Conveyance systems (pruning of fruit plants are management practices which apply technical criteria for productivity in temperate climates producing countries. It is known in Boyacá, except traditional open vessel driving and other forms of effects, which is why the study was to evaluate six different types of driving on the vegetative growth of two peach varieties. We measured plant height, stem diameter, length of primary branches, percentage of lignification, and canopy projection based on each form (treatment. Was observed that the different types of pruning positively influenced with respect to the absolute control in the vegetative growth of the plants and the lignification of the primary branches, with the highest incidence in the range Rubidoux. The results also showed that the more a tree height growth and lignification of primary branches was husito.

  4. Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions. (United States)

    Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie


    Due to their remarkable resistance to extreme conditions, Deinococcaceae strains are of great interest to biotechnological prospects. However, the physiology of the extremophile strain Deinococcus geothermalis has scarcely been studied and is not well understood. The physiological behaviour was then studied in well-controlled conditions in flask and bioreactor cultures. The growth of D. geothermalis type strains was compared. Among the strains tested, the strain from the German Collection of Microorganisms (Deutsche Sammlung von Mikroorganismen DSM) DSM-11302 was found to give the highest biomass concentration and growth rate: in a complex medium with glucose, the growth rate reached 0.75 h(-1) at 45 °C. Yeast extract concentration in the medium had significant constitutive and catalytic effects. Furthermore, the results showed that the physiological descriptors were not affected by the inoculum preparation steps. A batch culture of D. geothermalis DSM-11302 on defined medium was carried out: cells grew exponentially with a maximal growth rate of 0.28 h(-1) and D. geothermalis DSM-11302 biomass reached 1.4 g·L(-1) in 20 h. Then, 1.4 gDryCellWeight of biomass (X) was obtained from 5.6 g glucose (Glc) consumed as carbon source, corresponding to a yield of 0.3 CmolX·CmolGlc(-1); cell specific oxygen uptake and carbon dioxide production rates reached 216 and 226 mmol.CmolX(-1)·h(-1), respectively, and the respiratory quotient (QR) value varied from 1.1 to 1.7. This is the first time that kinetic parameters and yields are reported for D. geothermalis DSM-11302 grown on a mineral medium in well-controlled batch culture.

  5. Single Cell Protein Production by Saccharomyces cerevisiae Using an Optimized Culture Medium Composition in a Batch Submerged Bioprocess. (United States)

    Hezarjaribi, Mehrnoosh; Ardestani, Fatemeh; Ghorbani, Hamid Reza


    Saccharomyces cerevisiae PTCC5269 growth was evaluated to specify an optimum culture medium to reach the highest protein production. Experiment design was conducted using a fraction of the full factorial methodology, and signal to noise ratio was used for results analysis. Maximum cell of 8.84 log (CFU/mL) was resulted using optimized culture composed of 0.3, 0.15, 1, and 50 g L(-1) of ammonium sulfate, iron sulfate, glycine, and glucose, respectively at 300 rpm and 35 °C. Glycine concentration (39.32 % contribution) and glucose concentration (36.15 % contribution) were determined as the most effective factors on the biomass production, while Saccharomyces cerevisiae growth had showed the least dependence on ammonium sulfate (5.2 % contribution) and iron sulfate (19.28 % contribution). The most interaction was diagnosed between ammonium sulfate and iron sulfate concentrations with interaction severity index of 50.71 %, while the less one recorded for glycine and glucose concentration was equal to 8.12 %. An acceptable consistency of 84.26 % was obtained between optimum theoretical cell numbers determined by software of 8.91 log (CFU/mL), and experimentally measured one at optimal condition confirms the suitability of the applied method. High protein content of 44.6 % using optimum culture suggests that Saccharomyces cerevisiae is a good commercial case for single cell protein production.

  6. Robust optimization for nonlinear time-delay dynamical system of dha regulon with cost sensitivity constraint in batch culture (United States)

    Yuan, Jinlong; Zhang, Xu; Liu, Chongyang; Chang, Liang; Xie, Jun; Feng, Enmin; Yin, Hongchao; Xiu, Zhilong


    Time-delay dynamical systems, which depend on both the current state of the system and the state at delayed times, have been an active area of research in many real-world applications. In this paper, we consider a nonlinear time-delay dynamical system of dha-regulonwith unknown time-delays in batch culture of glycerol bioconversion to 1,3-propanediol induced by Klebsiella pneumonia. Some important properties and strong positive invariance are discussed. Because of the difficulty in accurately measuring the concentrations of intracellular substances and the absence of equilibrium points for the time-delay system, a quantitative biological robustness for the concentrations of intracellular substances is defined by penalizing a weighted sum of the expectation and variance of the relative deviation between system outputs before and after the time-delays are perturbed. Our goal is to determine optimal values of the time-delays. To this end, we formulate an optimization problem in which the time delays are decision variables and the cost function is to minimize the biological robustness. This optimization problem is subject to the time-delay system, parameter constraints, continuous state inequality constraints for ensuring that the concentrations of extracellular and intracellular substances lie within specified limits, a quality constraint to reflect operational requirements and a cost sensitivity constraint for ensuring that an acceptable level of the system performance is achieved. It is approximated as a sequence of nonlinear programming sub-problems through the application of constraint transcription and local smoothing approximation techniques. Due to the highly complex nature of this optimization problem, the computational cost is high. Thus, a parallel algorithm is proposed to solve these nonlinear programming sub-problems based on the filled function method. Finally, it is observed that the obtained optimal estimates for the time-delays are highly satisfactory

  7. Improvement of cloned [alpha]-amylase gene expression in fed-batch culture of recombinant Saccharomyces cerevisiae by regulating both glucose and ethanol concentrations using a fuzzy controller

    Energy Technology Data Exchange (ETDEWEB)

    Shiba, Sumihisa; Nishida, Yoshio; Park, Y.S.; Iijima, Shinji; Kobayashi, Takeshi (Nagoya Univ. (Japan). Dept. of Biotechnology)


    The effect of ethanol concentration on cloned gene expression in recombinant Saccharomyces cerevisiae strain 20B-12 containing one of two plasmids, pNA3 and pNA7, was investigated in batch cultures. Plasmids pNA3 and pNA7 contain the [alpha]-amylase gene under the control of the SUC2 or PGK promoter, respectively. When the ethanol concentration was controlled at 2 to 5 g/L, the gene expressions were two times higher than those at 20 g/L ethanol. To increase the gene expression by maintaining both the ethanol and glucose concentrations at low levels, a fuzzy controller was developed. The concentrations of glucose and ethanol were controlled simultaneously at 0.15 and 2 g/L, respectively, in the production phase using the fuzzy controller in fed-batch culture. The synthesis of [alpha]-amylase was induced by the low glucose concentration and maintained at a high level of activity by regulating the ethanol concentration at 2 g/L. The secretory [alpha]-amylase activities of cells harboring plasmids pNA3 and pNA7 in fed-batch culture were 175 and 392 U/mL, and their maximal specific activities 7.7 and 12.4 U/mg dry cells, respectively. These values are two to three times higher in activity and three to four times higher in specific activity than those obtained when glucose only was controlled.

  8. Determination of some significant batch culture conditions affecting acetyl-xylan esterase production by Penicillium notatum NRRL-1249

    Directory of Open Access Journals (Sweden)

    Akhtar MN


    Full Text Available Abstract Background Acetyl-xylan esterase (AXE, EC hydrolyses acetate group from the linear chain of xylopyranose residues bound by β-1,4-linkage. The enzyme finds commercial applications in bio-bleaching of wood pulp, treating animal feed to increase digestibility, processing food to increase clarification and converting lignocellulosics to feedstock and fuel. In the present study, we report on the production of an extracellular AXE from Penicillium notatum NRRL-1249 by solid state fermentation (SSF. Results Wheat bran at a level of 10 g (with 4 cm bed height was optimized as the basal substrate for AXE production. An increase in enzyme activity was observed when 7.5 ml of mineral salt solution (MSS containing 0.1% KH2PO4, 0.05% KCl, 0.05% MgSO4.7H2O, 0.3% NaNO3, 0.001% FeSO4.2H2O and 0.1% (v/w Tween-80 as an initial moisture content was used. Various nitrogen sources including ammonium sulphate, urea, peptone and yeast extract were compared for enzyme production. Maximal enzyme activity of 760 U/g was accomplished which was found to be highly significant (p ≤ 0.05. A noticeable enhancement in enzyme activity was observed when the process parameters including incubation period (48 h, initial pH (5, 0.2% (w/w urea as nitrogen source and 0.5% (v/w Tween-80 as a stimulator were further optimized using a 2-factorial Plackett-Burman design. Conclusion From the results it is clear that an overall improvement of more than 35% in terms of net enzyme activity was achieved compared to previously reported studies. This is perhaps the first report dealing with the use of P. notatum for AXE production under batch culture SSF. The Plackett-Burman model terms were found highly significant (HS, suggesting the potential commercial utility of the culture used (df = 3, LSD = 0.126.

  9. Stochastic modelling of Listeria monocytogenes single cell growth in cottage cheese with mesophilic lactic acid bacteria from aroma producing cultures

    DEFF Research Database (Denmark)

    Østergaard, Nina Bjerre; Christiansen, Lasse Engbo; Dalgaard, Paw


    A stochastic model was developed for simultaneous growth of low numbers of Listeria monocytogenes and populations of lactic acid bacteria from the aroma producing cultures applied in cottage cheese. During more than two years, different batches of cottage cheese with aroma culture were analysed...... for pH, lactic acid concentration and initial concentration of lactic acid bacteria. These data and bootstrap sampling were used to represent product variability in the stochastic model. Lag time data were estimated from observed growth data (lactic acid bacteria) and from literature on L. monocytogenes....... 2014. Modelling the effect of lactic acid bacteria from starter- and aroma culture on growth of Listeria monocytogenes in cottage cheese. International Journal of Food Microbiology. 188, 15-25]. Growth of L. monocytogenes single cells, using lag time distributions corresponding to three different...

  10. Quantitative Polymerase Chain Reaction for Microbial Growth Kinetics of Mixed Culture System. (United States)

    Cotto, Ada; Looper, Jessica K; Mota, Linda C; Son, Ahjeong


    Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μmax and Ks). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

  11. 分批补料及缺氮培养对小球藻油脂产量的影响%Effects of Fed-batch and Nitrogen-deficient Culture on Lipid Yield of Chlorella sp.

    Institute of Scientific and Technical Information of China (English)

    葛珍珍; 王杰; 余晓斌


    目的 为了实现小球藻的高密度及高产油培养。方法 通过分析分批培养过程中藻细胞的生长曲线,葡萄糖消耗曲线,pH及溶氧变化曲线,以小球藻进行分批补料,待藻细胞达到一定的高密度后再进行缺氮培养以富集细胞内的油脂。结果 经过4次分批补料,小球藻的生物量达到了65.25g/L,然后进行缺氮培养12h,然后进行缺氮培养12h,小球藻的油脂含量由42.75%提高到63.82%,油脂含量达43.47g/L.结论 合理的分批补料明显地提高了小球藻的生物量。缺氮培养进一步提高了小球藻的油脂含量。%[Objective] This study was to realize high-density culture of Chlorella sp. as well as the culture with high lipid yield. [Method] Through analyzing the growth curve of Chlorella sp. cells, dextrose consumption curve, change curves of pH and dissolved oxygen, a fed-batch culture was conducted, followed by a nitrogen-deficient culture aiming at accumulating the lipids in Chlorella sp. cells when a high density of Chlorella sp. cells was obtained. [Result] After four batches of feeding were pro- vided, the biomass of Chlorella sp. reached up to 65.25 g/L, and the lipid content increased from 42.75% to 63.82% in Chlorella sp. cells, with the yield of 43.37 g/L in the following 12 hours of nitrogen-deficient culture. [Conclusion] Reasonable fed- batch can significantly improve the biomass of Chlorella sp., and the nitrogen-defi- cient culture further raises the lipid yield of Chlorella sp.

  12. H(2) production through anaerobic mixed culture: effect of batch S(0)/X(0) and shock loading in CSTR. (United States)

    Fan, Kuo-Shuh; Chen, Ya-Yun


    Biological production of H(2) has received considerable attention lately. The present study was undertaken to observe the effects of substrate/seeding ratios (S(0)/X(0)) on batch H(2) generation. The H(2)-producing seeding spores were obtained from the heat treatment (88 degrees C for 12h) of the compost from a grass composting facility. A dehydrated brewery mixture was used as feed substrate. The results indicate that the pattern of the cumulative H(2) production with time is similar to the growth curve with a typical lag, exponential and stationary phase; the results were successfully modeled with a modified Gompertz equation. It appears that maximum H(2) yield potential (27ml g(-1)COD(added)) occurs at an S(0)/X(0) ratio of about 4, whereas the maximum specific H(2) yield (205ml g(-1) VSSd(-1)) occurs at approximately S(0)/X(0)=3. The S(0)/X(0) ratios higher than 4 would inhibit H(2) production. An attempt was made to waste a certain amount of reactor content and replaced it with fresh substrate in order to enhance H(2) production. After this medium replacement, the H(2) production was initially inhibited and the system then exhibited a long lag before it reached an active H(2) production stage. For a continuous-stirred tank-reactor (CSTR) system, the results of replacing 25% of the reactor content indicate that there is still a lag time before a sudden increase in H(2) production after the addition of the new substrate feed. The major low molecular weight acids identified are HAc and HBu with total volatile acids of about 6000-8000mg l(-1). The ratio of HAc/HBu in the present study is relatively constant (about 5) and appears not significantly affected by the medium replacement. The concentration of total alcohols is about 2000mg l(-1). All in all, the CSTR system is able to recover to its previous performance after such a dramatic 25% medium replacement.

  13. Promoting Teacher Growth through Lesson Study: A Culturally Embedded Approach (United States)

    Ebaeguin, Marlon


    Lesson Study has captured the attention of many international educators with its promise of improved student learning and sustained teacher growth. Lesson Study, however, has cultural underpinnings that a simple transference model overlooks. A culturally embedded approach attends to the existing cultural orientations and values of host schools.…

  14. Improved elastase production by Bacillus sp.EL31410--further optimization and kinetics studies of culture medium for batch fermentation

    Institute of Scientific and Technical Information of China (English)

    何国庆; 陈启和; 琚晓捷; 石乃冬


    An efficient culture medium producing a bacterial elastase with high yields was developed further following preliminary studies by means of response surface method. Central composite design (CCD) and response surface methodology were applied to optimize the medium constituents. A central composite design was used to explain the combined effect of three medium constituents, viz, glucose, K2HPO4, MgSO4@7H2O. The strain produced more elastase in the completely optimized medium, as compared with the partially optimized medium. The fitted model of the second model, as per RSM,showed that glucose was 7.4 g/100 ml, casein 1.13 g/100 ml, corn steep flour 0.616 g/100 ml, K2HPO4 0.206 g/100 ml and MgSO4@7H2O 0.034 g/100 ml. The fermentation kinetics of these two culture media in the flask experiments were analyzed. It was found that the highest elastase productivity occurred at 54 hours. Higher glucose concentration had inhibitory effect on elastase production. At the same time, we observed that the glucose consumption rate was slow in the completely optimized medium, which can explain the lag period of the highest elastase production. Some metal ions and surfactant additives also affected elastase production and cell growth.

  15. Impact of the freeze-drying process on product appearance, residual moisture content, viability, and batch uniformity of freeze-dried bacterial cultures safeguarded at culture collections. (United States)

    Peiren, Jindrich; Hellemans, Ann; De Vos, Paul


    In this study, causes of collapsed bacterial cultures in glass ampoules observed after freeze-drying were investigated as well as the influence of collapse on residual moisture content (RMC) and viability. Also, the effect of heat radiation and post freeze-drying treatments on the RMC was studied. Cake morphologies of 21 bacterial strains obtained after freeze-drying with one standard protocol could be classified visually into four major types: no collapse, porous, partial collapse, and collapse. The more pronounced the collapse, the higher residual moisture content of the freeze-dried product, ranging from 1.53 % for non-collapsed products to 3.62 % for collapsed products. The most important cause of collapse was the mass of the inserted cotton plug in the ampoule. Default cotton plugs with a mass between 21 and 30 mg inside the ampoule did not affect the viability of freeze-dried Aliivibrio fischeri LMG 4414(T) compared to ampoules without cotton plugs. Cotton plugs with a mass higher than 65 mg inside the ampoule induced a full collapsed product with rubbery look (melt-back) and decreasing viability during storage. Heat radiation effects in the freeze-drying chamber and post freeze-drying treatments such as exposure time to air after freeze-drying and manifold drying time prior to heat sealing of ampoules influenced the RMC of freeze-dried products. To produce uniform batches of freeze-dried bacterial strains with intact cake structures and highest viabilities, inserted cotton plugs should not exceed 21 mg per ampoule. Furthermore, heat radiation effects should be calculated in the design of the primary drying phase and manifold drying time before heat sealing should be determined as a function of exposure time to air.

  16. The rules on suspension cell batch culture of Hedyotis diffusa%白花蛇舌草悬浮细胞分批培养规律的研究

    Institute of Scientific and Technical Information of China (English)

    于瑞莲; 许金国; 顾晓娟


    rate of [ NO3- ] is far higher than [ NH4+ ] ,to 7. 14 μg/( mL ? d) ,cell growth/and the polysaccharide production were non-coupled and the polysaccharide was generated in the late culture stage. Conclusion It was found that the rules on suspension cell batch culture of Hedyotis diffusa with the time is helpful for later culture technique.

  17. Growth of Geobacter sulfurreducens under nutrient-limiting conditions in continuous culture. (United States)

    Esteve-Núñez, Abraham; Rothermich, Mary; Sharma, Manju; Lovley, Derek


    A system for growing Geobacter sulfurreducens under anaerobic conditions in chemostats was developed in order to study the physiology of this organism under conditions that might more closely approximate those found in the subsurface than batch cultures. Geobacter sulfurreducens could be cultured under acetate-limiting conditions with fumarate or Fe(III)-citrate as the electron acceptor at growth rates between 0.04 and 0.09 h(-1). The molar growth yield was threefold higher with fumarate as the electron acceptor than with Fe(III), despite the lower mid-point potential of the fumarate/succinate redox couple. When growth was limited by availability of fumarate, high steady-state concentrations were detected, suggesting that fumarate is unlikely to be an important electron acceptor in sedimentary environments. The half-saturation constant, Ks, for acetate in Fe(III)-grown cultures (10 microM) suggested that the growth of Geobacter species is likely to be acetate limited in most subsurface sediments, but that when millimolar quantities of acetate are added to the subsurface in order to promote the growth of Geobacter for bioremediation applications, this should be enough to overcome any acetate limitations. When the availability of electron acceptors, rather than acetate, limited growth, G. sulfurreducens was less efficient in incorporating acetate into biomass but had higher respiration rates, a desirable physiological characteristic when adding acetate to stimulate the activity of Geobacter species during in situ uranium bioremediation. These results demonstrate that the ability to study the growth of G. sulfurreducens under steady-state conditions can provide insights into its physiological characteristics that have relevance for its activity in a diversity of sedimentary environments.

  18. Growth and activity of Bulgarian yogurt starter culture in iron-fortified milk. (United States)

    Simova, Emilina; Ivanov, Galin; Simov, Zhelyazko


    Bulgarian yogurts were manufactured and fortified with 8, 15 and 27 mg of iron kg(-1) of yogurt. The growth and acidifying activity of the starter culture bacteria Streptococcus thermophilus 13a and Lactobacillus delbrueckii subsp. bulgaricus 2-11 were monitored during milk fermentation and over 15 days of yogurt storage at 4 degrees C. Fortifying milk with iron did not affect significantly the growth of the starter culture during manufacture and storage of yogurt. Counts of yogurt bacteria at the end of fermentation of iron-fortified milks were between 2.1 x 10(10) and 4.6 x 10(10) CFU ml(-1), which were not significantly different from numbers in unfortified yogurts. In all batches of yogurt, the viable cell counts of S. thermophilus 13a were approximately three times higher than those of L. delbrueckii subsp. bulgaricus 2-11. Greater decrease in viable cell count over 15 days of storage was observed for S. thermophilus 13a compared to L. delbrueckii subsp. bulgaricus 2-11. Intensive accumulation of lactic acid was observed during incubation of milk and all batches reached pH 4.5 +/- 0.1 after 3.0 h. At the end of fermentation process, lactic acid concentrations in iron-fortified yogurts were between 6.9 +/- 0.4 and 7.3 +/- 0.5 g l(-1). The acidifying activity of starter culture bacteria in the control and iron-fortified milks was similar. There was no increase in oxidized, metallic and bitter off-flavors in iron-fortified yogurts compared to the control. Iron-fortified yogurts did not differ significantly in their sensorial, chemical and microbiological characteristics with unfortified yogurt, suggesting that yogurt is a suitable vehicle for iron fortification and that the ferrous lactate is an appropriate iron source for yogurt fortification.


    Directory of Open Access Journals (Sweden)

    Abdullah Mochamad Busairi


    Full Text Available Pineapple juice waste contains valuable components, which are mainly sucrose, glucose, and fructose. Recently, lactic acid has been considered to be an important raw material for the production of biodegradable lactide polymer. The fermentation experiments were carried out in a 3 litres fermentor (Biostat B Model under anaerobic condition with stirring speed of 50 rpm, temperature at 40oC, and pH of 6.00. Effect of feed concentration on lactic acid production, bacterial growth, substrate utilisation and productivity was studied. The results obtained from fed- batch culture fermentation showed that the maximum lactic acid productivity was 0.44 g/L.h for feed concentration of 90 g/L at 48 hours. Whereas the lactic acid productivity obtained from fed-batch culture was twice and half fold higher than that of batch culture productivity.  Buangan jus nanas mengandung komponen yang berharga terutama sukrosa, glukosa, dan fruktosa. Asam laktat adalah bahan baku yang terbaru dan penting untuk dibuat sebagai polimer laktat yang dapat terdegradasi oleh lingkungan. Percobaan dilakukan pada fermentor 3 liter (Model Biostat B di bawah kondisi anaerob dengan kecepatan pengadukan 50 rpm, temperatur 40oC, dan pH 6,00. Pengaruh konsentrasi umpan terhadap produksi asam laktat, pertumbuhan mikroba, pengggunaan substrat dan produktivitas telah dipelajari. Hasil yang didapatkan pada fermentasi dengan menggunakan sistem fed-batch menunjukkan bahwa produktivitas asam laktat maksimum adalah 0.44 g/L,jam dengan konsentrasi umpan, 90 g/L pada waktu 48 jam. Bahkan produktivitas asam laktat yang didapat pada kultur fed-batch lebih tinggi 2,5 kali dari pada proses menggunakan sistem batch

  20. Hydrogen production from starch by co-culture of Clostridium acetobutylicum and Rhodobacter sphaeroides in one step hybrid dark- and photofermentation in repeated fed-batch reactor. (United States)

    Zagrodnik, R; Łaniecki, M


    Hydrogen production from starch by a co-culture hybrid dark and photofermentation under repeated fed-batch conditions at different organic loading rates (OLR) was studied. Effective cooperation between bacteria in co-culture during initial days was observed at controlled pH 7.0. However, at pH above 6.5 dark fermentation phase was redirected from H2 formation towards production of formic acid, lactic acid and ethanol (which are not coupled with hydrogen production) with simultaneous lower starch removal efficiency. This resulted in decrease in the hydrogen production rate. The highest H2 production in co-culture process (3.23LH2/Lmedium - after 11days) was achieved at OLR of 1.5gstarch/L/day, and it was twofold higher than for dark fermentation process (1.59LH2/Lmedium). The highest H2 yield in the co-culture (2.62molH2/molhexose) was obtained at the OLR of 0.375gstarch/L/day. Different pH requirements of bacteria were proven to be a key limitation in co-culture system.

  1. Changes In Growth Culture FDA Activity Under Changing Growth Conditions

    DEFF Research Database (Denmark)

    Jørgensen, Per Elberg; Eriksen, Thomas Juul; Jensen, Bjørn K.


    of the bacteria. The FDA activity/ATP ratio was calculated for different concentrations of autoclaved sludge. A faster decay rate of ATP relative to FDA hydrolysis activity was observed, thus causing changes in the ratio. Furthermore, comparison between values obtained from pure cultures and different soils...

  2. Microalgal Photosynthesis and Growth in Mass Culture

    NARCIS (Netherlands)

    Janssen, M.G.J.


    The development of large-scale outdoor microalgae production requires a thorough understanding of microalgal growth which should be encompassed in a mathematical model. The model should be as simple as possible allowing use in outdoor practice by persons with varying backgrounds. This chapter provid

  3. Mineralization and growth of cultured embryonic skeletal tissue in microgravity (United States)

    Klement, B. J.; Spooner, B. S.


    Microgravity provides a unique environment in which to study normal and pathological phenomenon. Very few studies have been done to examine the effects of microgravity on developing skeletal tissue such as growth plate formation and maintenance, elongation of bone primordia, or the mineralization of growth plate cartilage. Embryonic mouse premetatarsal triads were cultured on three space shuttle flights to study cartilage growth, differentiation, and mineralization, in a microgravity environment. The premetatarsal triads that were cultured in microgravity all formed cartilage rods and grew in length. However, the premetatarsal cartilage rods cultured in microgravity grew less in length than the ground control cartilage rods. Terminal chondrocyte differentiation also occurred during culture in microgravity, as well as in the ground controls, and the matrix around the hypertrophied chondrocytes was capable of mineralizing in both groups. The same percentage of premetatarsals mineralized in the microgravity cultures as mineralized in the ground control cultures. In addition, the sizes of the mineralized areas between the two groups were very similar. However, the amount of 45Ca incorporated into the mineralized areas was significantly lower in the microgravity cultures, suggesting that the composition or density of the mineralized regions was compromised in microgravity. There was no significant difference in the amount of 45Ca liberated from prelabeled explants in microgravity or in the ground controls.

  4. Biosynthesis of highly enriched 13C-lycopene for human metabolic studies using repeated batch tomato cell culturing with 13C-glucose. (United States)

    Moran, Nancy Engelmann; Rogers, Randy B; Lu, Chi-Hua; Conlon, Lauren E; Lila, Mary Ann; Clinton, Steven K; Erdman, John W


    While putative disease-preventing lycopene metabolites are found in both tomato (Solanum lycopersicum) products and in their consumers, mammalian lycopene metabolism is poorly understood. Advances in tomato cell culturing techniques offer an economical tool for generation of highly-enriched (13)C-lycopene for human bioavailability and metabolism studies. To enhance the (13)C-enrichment and yields of labelled lycopene from the hp-1 tomato cell line, cultures were first grown in (13)C-glucose media for three serial batches and produced increasing proportions of uniformly labelled lycopene (14.3±1.2%, 39.6±0.5%, and 48.9±1.5%) with consistent yields (from 5.8 to 9 mg/L). An optimised 9-day-long (13)C-loading and 18-day-long labelling strategy developed based on glucose utilisation and lycopene yields, yielded (13)C-lycopene with 93% (13)C isotopic purity, and 55% of isotopomers were uniformly labelled. Furthermore, an optimised acetone and hexane extraction led to a fourfold increase in lycopene recovery from cultures compared to a standard extraction.

  5. Differences in carbohydrate profiles in batch culture grown planktonic and biofilm cells of Amphora rostrata Wm. Sm

    Digital Repository Service at National Institute of Oceanography (India)

    Khodse, V.B.; Bhosle, N.B.

    modes of growth, the concentration of total carbohydrates, carbohydrate fractions, neutral carbohydrates, uronic acids and amino sugars in planktonic and biofilm cells of Amphora rostrata were measured. The results showed that the distribution...

  6. Mechanically induced alterations in cultured skeletal muscle growth (United States)

    Vandenburgh, H. H.; Hatfaludy, S.; Karlisch, P.; Shansky, J.


    Model systems are available for mechanically stimulating cultured skeletal muscle cells by passive tensile forces which simulate those found in vivo. When applied to embryonic muscle cells in vitro these forces induce tissue organogenesis, metabolic adaptations, and muscle cell growth. The mechanical stimulation of muscle cell growth correlates with stretch-induced increases in the efflux of prostaglandins PGE2 and PGF2(alpha) in a time and frequency dependent manner. These prostaglandins act as mechanical 'second messengers' regulating skeletal muscle protein turnover rates. Since they also effect bone remodelling in response to tissue loading and unloading, secreted prostaglandins may serve as paracrine growth factors, coordinating the growth rates of muscle and bone in response to external mechanical forces. Cell culture model systems will supplement other models in understanding mechanical transduction processes at the molecular level.

  7. Microspectroscopic investigation of the membrane clogging during the sterile filtration of the growth media for mammalian cell culture. (United States)

    Cao, Xiaolin; Loussaert, James A; Wen, Zai-qing


    Growth media for mammalian cell culture are very complex mixtures of several dozens of ingredients, and thus the preparation of qualified media is critical to viable cell density and final product titers. For liquid media prepared from powdered ingredients, sterile filtration is required prior to use to safeguard the cell culture process. Recently one batch of our prepared media failed to pass through the sterile filtration due to the membrane clogging. In this study, we report the root cause analysis of the failed sterile filtration based on the investigations of both the fouling media and the clogged membranes with multiple microspectroscopic techniques. Cellular particles or fragments were identified in the fouling media and on the surfaces of the clogged membranes, which were presumably introduced to the media from the bacterial contamination. This study demonstrated that microspectroscopic techniques may be used to rapidly identify both microbial particles and inorganic precipitates in the cell culture media.

  8. Reduction of excess sludge in a sequencing batch reactor by lysis-cryptic growth using quick lime for disintegration under low temperature. (United States)

    Lv, Xiao-Mei; Song, Ju-Sheng; Li, Ji; Zhai, Kun


    In the present study, quick-lime-based thermal-alkaline sludge disintegration (SD) under low temperature was combined with cryptic growth to investigate the excess sludge reduction efficiency in the sequencing batch reactor (SBR). The optimized condition of SD was as follows: T = 80℃, pH = 11, t = 180 min, and the SD rate was about 42.1%. With 65.6% of excess sludge disintegrated and returned to the SBR, the system achieved sludge reduction rate of about 40.1%. The lysis-cryptic growth still obtained satisfactory sludge reduction efficiency despite the comparative low SD rate, which suggested that disintegration rate might not be the decisive factor for cryptic-growth-based sludge reduction. Lysis-cryptic growth did not impact the effluent quality, yet the phosphorus removal performance was enhanced, with effluent total phosphorus concentration decreased by 0.3 mg/L (33%). Crystal compounds of calcium phosphate precipitate were detected in the system by Fourier transform infrared spectroscopy and X-ray diffraction, which indicated the phosphorus removal potential of SD using lime. Moreover, endogenous dehydrogenase activity of activated sludge in the lysis-cryptic system was enhanced, which was beneficial for sludge reduction. SD and cryptic growth in the present study demonstrates an economical and effective approach for sludge reduction.

  9. Improvement of Growth and Periplocin Yield of Periploca sepium Adventitious Root Cultures by Altering Nitrogen Source Supply

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jian; GAO Wen-yuan; WANG Juan; LI Xing-lin; XIAO Pei-gen


    Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH-NO-)at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH-NO(1:2)and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.

  10. Growth and Plating of Cell Suspension Cultures of Datura Innoxia

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen


    Suspension cultures of Datura innoxia Mill, were successfully grown on a modified Murashige and Skoog medium with 2,4–D, NAA or BAP as growth substances, provided the micronutrient levels were reduced to 1/10. Normal amounts of micronutrients were toxic. Attempts to identify the toxic elements did...

  11. Growth and Cultural Characteristics of Cordyceps cardinalis Collected from Korea. (United States)

    Sung, Gi-Ho; Shrestha, Bhushan; Han, Sang-Kuk; Kim, Soo-Young; Sung, Jae-Mo


    Cordyceps cardinalis was reported in Japan and the USA in 2004, and its fruiting bodies have recently been cultured in Korea. Herbarium specimens preserved at the Cordyceps Research Institute, Mushtech, Korea were revised and identified as C. cardinalis, based on morphological characters and conidial structures. Most of the C. cardinalis specimens were collected from Mt. Halla in Jeju-do. The effects of various nutritional sources and environmental conditions such as temperature and pH on mycelial growth of C. cardinalis were studied. Oatmeal agar, Martin's peptone dextrose agar, and Schizophyllum (mushroom) genetics complete medium plus yeast extract resulted in the best mycelial growth. Among carbon sources, cereals, and nitrogen sources, maltose, oatmeal, and peptone resulted in the best mycelial growth respectively. Mineral salts helped to increase growth rate but only resulted in thin mycelial density, similar to water agar. A temperature of 25℃ and a pH of 7 resulted in the highest mycelial growth. Based on these results, a Cordyceps cardinalis composite medium (CCM) was formulated with 1% maltose, 2% oatmeal, 1% peptone, and 2% agar. Use of the CCM resulted in slightly better mycelial growth than that of other commonly used agar media. Only organic nitrogen sources imparted a reddish pigmentation to the agar media, but this character diminished after several subcultures. A 7 day culture duration resulted in the best mycelial growth.

  12. Growth and Cultural Characteristics of Cordyceps cardinalis Collected from Korea (United States)

    Sung, Gi-Ho; Shrestha, Bhushan; Han, Sang-Kuk; Kim, Soo-Young


    Cordyceps cardinalis was reported in Japan and the USA in 2004, and its fruiting bodies have recently been cultured in Korea. Herbarium specimens preserved at the Cordyceps Research Institute, Mushtech, Korea were revised and identified as C. cardinalis, based on morphological characters and conidial structures. Most of the C. cardinalis specimens were collected from Mt. Halla in Jeju-do. The effects of various nutritional sources and environmental conditions such as temperature and pH on mycelial growth of C. cardinalis were studied. Oatmeal agar, Martin's peptone dextrose agar, and Schizophyllum (mushroom) genetics complete medium plus yeast extract resulted in the best mycelial growth. Among carbon sources, cereals, and nitrogen sources, maltose, oatmeal, and peptone resulted in the best mycelial growth respectively. Mineral salts helped to increase growth rate but only resulted in thin mycelial density, similar to water agar. A temperature of 25℃ and a pH of 7 resulted in the highest mycelial growth. Based on these results, a Cordyceps cardinalis composite medium (CCM) was formulated with 1% maltose, 2% oatmeal, 1% peptone, and 2% agar. Use of the CCM resulted in slightly better mycelial growth than that of other commonly used agar media. Only organic nitrogen sources imparted a reddish pigmentation to the agar media, but this character diminished after several subcultures. A 7 day culture duration resulted in the best mycelial growth. PMID:23956666

  13. 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


    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.

  14. Combined effect of the methanol utilization (Mut) phenotype and gene dosage on recombinant protein production in Pichia pastoris fed-batch cultures. (United States)

    Cos, Oriol; Serrano, Alicia; Montesinos, José Luis; Ferrer, Pau; Cregg, James M; Valero, Francisco


    An important number of heterologous proteins have been produced in the methylotrophic yeast Pichia pastoris using the alcohol oxidase promoter. Two factors that drastically influence protein production and cultivation process development in this system are gene dosage and methanol assimilation capacity of the host strain (Mut phenotype). Using a battery of four strains which secrete a Rhizopus oryzae lipase (ROL), the combined effects of gene dosage and Mut phenotype on recombinant protein production in Pichia pastoris was studied in fed-batch cultures. Regarding the effect of phenotype, the specific productivity and the Y(P/X) were 1.29- and 2.34-fold higher for Mut(s)ROL single copy strain than for Mut+ROL single copy strain. On the contrary, the productivity of Mut+ROL single copy strain was 1.34-fold higher than Mut(s)ROL single copy strain. An increase in ROL gene dosage seems to negatively affect cell's performance in bioreactor cultures, particularly in Mut(s) strains. Overall, the Mut(s) strain may be still advantageous to use because it allows for easier process control strategies.

  15. Production of gymnemic acid depends on medium, explants, PGRs, color lights, temperature, photoperiod, and sucrose sources in batch culture of Gymnema sylvestre. (United States)

    Ahmed, A Bakrudeen Ali; Rao, A S; Rao, M V; Taha, Rosna Mat


    Gymnema sylvestre (R.Br.) is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA). The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L) and KN (0.5 mg/L). Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w) was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w). Maximum GA production (58.28 mg/g d.w) was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures of G. sylvestre. The factors optimized for in vitro production of GA during the present study can successfully be employed for their large-scale production in bioreactors.

  16. Batch culture fermentation of Penicillium chrysogenum and a report on the isolation, purification, identification and antibiotic activity of citrinin

    Digital Repository Service at National Institute of Oceanography (India)

    PrabhaDevi; DeSouza, L.; Kamat, T.; Rodrigues, C.; Naik, C.G.

    is reported to be a broad spectrum antibiotic especially against Gram positive-bacteria. The present study consists report on Gram-positive bacteria, its activity against Gram-negative bacteria and on its antifungal properties. Primarily, citrinin is a...). Previous Fig. 1—Time course of biomass accumulation and citrinin production during growth of Penicillium chrysogenum in PDB medium. Table 1—Antibacterial activity of citrinin Bacterial pathogens Gram stain Streptomycin (100µg.disc-1...

  17. Stochastic modelling of Listeria monocytogenes single cell growth in cottage cheese with mesophilic lactic acid bacteria from aroma producing cultures. (United States)

    Østergaard, Nina Bjerre; Christiansen, Lasse Engbo; Dalgaard, Paw


    A stochastic model was developed for simultaneous growth of low numbers of Listeria monocytogenes and populations of lactic acid bacteria from the aroma producing cultures applied in cottage cheese. During more than two years, different batches of cottage cheese with aroma culture were analysed for pH, lactic acid concentration and initial concentration of lactic acid bacteria. These data and bootstrap sampling were used to represent product variability in the stochastic model. Lag time data were estimated from observed growth data (lactic acid bacteria) and from literature on L. monocytogenes single cells. These lag time data were expressed as relative lag times and included in growth models. A stochastic model was developed from an existing deterministic growth model including the effect of five environmental factors and inter-bacterial interaction [Østergaard, N.B, Eklöw, A and Dalgaard, P. 2014. Modelling the effect of lactic acid bacteria from starter- and aroma culture on growth of Listeria monocytogenes in cottage cheese. International Journal of Food Microbiology. 188, 15-25]. Growth of L. monocytogenes single cells, using lag time distributions corresponding to three different stress levels, was simulated. The simulated growth was subsequently compared to growth of low concentrations (0.4-1.0 CFU/g) of L. monocytogenes in cottage cheese, exposed to similar stresses, and in general a good agreement was observed. In addition, growth simulations were performed using population relative lag time distributions for L. monocytogenes as reported in literature. Comparably good predictions were obtained as for the simulations performed using lag time data for individual cells of L. monocytogenes. Therefore, when lag time data for individual cells are not available, it was suggested that relative lag time distributions for L. monocytogenes can be used as a qualified default assumption when simulating growth of low concentrations of L. monocytogenes.

  18. Manufacturing technology for high quality swept-culture growth quartz (United States)

    Balascio, J. F.; Lias, N. C.


    Relatively low impurity alpha-quartz has been grown from the hydroxide mineralizer system with the use of three types of nutrient: converted, special cultured and cultured. The converted nutrient resulted in the lowest total ppm level (2.6) and Al ppm level (0.5) in the as-grown crystals. Special cultured and cultured nutrients resulted in 1.7 ppm and 2.1 ppm of aluminum in crystals grown, respectively, from these nutrients. Infrared Q values ranged from 2.5x10 to the 6th power in these crystals. With respect to rho in the as-grown crystals, a ten minute etching of seeds in 7.0 molal ammonium bifluoride has resulted in the growth of the lowest etch channel density crystals (approx = 135/sq cm).

  19. Evaluation on prebiotic properties of β-glucan and oligo-β-glucan from mushrooms by human fecal microbiota in fecal batch culture

    Directory of Open Access Journals (Sweden)

    Chiraphon Chaikliang


    Full Text Available Background: β-glucan is dietary fiber, a structural polysaccharide, β-linked linear chains of D-glucose polymers with variable frequency of branches. β-glucan is isolated from different sources such as cell walls of baker’s yeast (Saccharomyces cerevisiae, cereals (oat and barley and various species of mushrooms. Among 8 mushrooms in the study, Schizophylum commune Fr and Auricularia auricula Judae had the highest in β-glucan contents and the cheapest cost of mushroom per content of β-glucan, respectively. Even the function of β-glucan on immune modulation has been known however no report on interaction between β-glucan and human gut microbiota. Gut microbiota is thought to have health effects by interaction with non-digestible component particular fermentable dietary fiber. It is important to correlate the specific groups of the microbial communities associated with β-glucan fermentation and the consequential SCFA profiles. β-glucan from mushroom may has potential prebiotic function similar to those from commercial yeast (Saccharomyces cerevisiae β-glucan. Objective: To evaluate on prebiotic properties of soluble β-glucans and oligo-β-glucans from Schizophylum commune Fr and Auricularia auricula Judae by fecal fermentation in batch culture. Methods: In vitro fecal fermentation in anaerobic batch cultures under simulated conditions similar to human colon with human faecal samples from three donors were performed. Comparison on 3 β-glucans and 2 oligo-β-glucans have been studied. Sample was taken at 0 h, 24 h and 48 h to analyze the numbers of bacterial changes by fluorescent in situ hybridization (FISH technique. Short chain fatty acids (SCFA were analyzed by HPLC. The prebiotic index (PI was calculated according to the change of 5 specific bacterial genus within 48 h fermentation. Results: Soluble β-glucan from Auricularia auricula Judae increased numbers of bifidobacteria and lactobacillus significantly (P<0.05. The PI of

  20. Model for the feedback control system of bacterial growth. II. Growth in continuous culture. (United States)

    Bleecken, S


    A mathematical model is developed that describes substrate limited bacterial growth in a continuous culture and that is based upon the conceptual framework elaborated in a previous paper for describing the feedback control system of cell growth [S. Bleecken, (1988). J. theor. Biol. 133, 37.] Central to the theory are the ideas that the limiting substrate is converted into low molecular weight building blocks of macromolecular synthesis which again are converted into biomass (RNA and protein) and that the rates of RNA and protein synthesis are controlled by the intracellular concentration of building blocks. It is shown that a continuous culture can be simulated by two interconnected feedback control systems the actuating signals of which are limiting substrate concentration and the intracellular concentration of building blocks, respectively. Three types of steady-states are found to appear in a continuous culture, besides the well-known stable steady-state of the whole culture there exist two batchlike steady-states of the biotic part of the culture which are metastable. The model is used to analyse the steady-states and their stability properties as well as the dynamic responses of biomass, RNA, protein, building block and substrate concentrations to changes in environmental conditions. Especially the inoculation of a continuous culture and the effects of step changes in dilution rate, inlet substrate concentration and growth temperature are studied in detail. Relations between the growth behaviour of a single cell and that of a continuous culture are derived. The RNA to protein ratio is introduced as a rough measure of the physiological state of cells and it is shown that a cell reacts to environmental changes with a simple pattern of basic responses in growth rate and physiological state. There are reasons to assume that the model presented is the minimal version of a structured model of bacterial growth and represents an optimum compromise between biological

  1. Detection of Sialic Acid-Utilising Bacteria in a Caecal Community Batch Culture Using RNA-Based Stable Isotope Probing

    Directory of Open Access Journals (Sweden)

    Wayne Young


    Full Text Available Sialic acids are monosaccharides typically found on cell surfaces and attached to soluble proteins, or as essential components of ganglioside structures that play a critical role in brain development and neural transmission. Human milk also contains sialic acid conjugated to oligosaccharides, glycolipids, and glycoproteins. These nutrients can reach the large bowel where they may be metabolised by the microbiota. However, little is known about the members of the microbiota involved in this function. To identify intestinal bacteria that utilise sialic acid within a complex intestinal community, we cultured the caecal microbiota from piglets in the presence of 13C-labelled sialic acid. Using RNA-based stable isotope probing, we identified bacteria that consumed 13C-sialic acid by fractionating total RNA in isopycnic buoyant density gradients followed by 16S rRNA gene analysis. Addition of sialic acid caused significant microbial community changes. A relative rise in Prevotella and Lactobacillus species was accompanied by a corresponding reduction in the genera Escherichia/Shigella, Ruminococcus and Eubacterium. Inspection of isotopically labelled RNA sequences suggests that the labelled sialic acid was consumed by a wide range of bacteria. However, species affiliated with the genus Prevotella were clearly identified as the most prolific users, as solely their RNA showed significantly higher relative shares among the most labelled RNA species. Given the relevance of sialic acid in nutrition, this study contributes to a better understanding of their microbial transformation in the intestinal tract with potential implications for human health.

  2. [High-level production of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) by feb-batch culture of Alcaligenes eutrophus]. (United States)

    Cai, Y B; Liu, M Q; Yi, Z H; Chen, Q; Weng, W Q


    Fermentation strategies for production P (3HB-co-3HV) from glucose and propionic (or valeric) acid by Alcaligenes eutrophus were studied. During the culture, we controlled pH of the broth by feeding precusors of 3HV- propionic or valeric acid after Ammonia feeding stopped. When propionic acid were used as the precusor, for 50 hours, we obtained a cell dry weight, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content and a 3HV fraction of 149.9 g/L, 124.9 g/L, 83.3% and 12.4 mol%, respectively, with a PHA productivity of 2.50 g h-1 L-1. When valeric acid were used as the precusor, for 45 hours, we obtained a cell dry weight, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content and a 3HV fraction of 160.2 g/L, 119.0 g/L, 74.2% and 17.7 mol%, respectively, with a PHA productivity of 2.64 g h-1 L-1. Prior to this study, it hasn't been reported to obtain such high level productivity and 3HV fraction at the same time by Alcaligenes eutrophus.

  3. 微量营养素对玉米酵母流加培养的影响%Effect ofmicronutrients on fed-batch culture of yeast on maize medium

    Institute of Scientific and Technical Information of China (English)

    李竹生; 张新伟; 宋娜; 牛芳方


    The effects of different amounts of micronutrients on the feeding rate of sugar solution, dissolved oxygen, yeast cell concentration and yeast cell growth during the fed-batch culture of yeast on maize medium were studied. It was showed that the optimal amount of the micronutrients were as follows: CaCl2 28.8mg/L, FeSO4 14.4mg/L, thiamine hydrochloride 4.8mg/L, biotin 0.12mg/L, D-calcium pantothenate 6.4mg/L and myoinositol 120mg/L. The yield of yeast had increase by 80.9% with the addition of micronutrients.%文中研究了微量营养素的不同添加量对玉米酵母流加培养过程中糖液流加速率、溶氧量、酵母细胞浓度、酵母细胞生长率的变化,得出微量营养素的添加量为CaCl2 28.8mg/L,FeSO4 14.4mg/L,盐酸硫氨4.Smg/L,生物素0.12mg/L,D-泛酸钙6.4mg/L,肌醇120mg/L,玉米酵母细胞的产率提高了80.9%.

  4. Identification of genes whose expressions are enhanced or reduced in baker's yeast during fed-batch culture process using molasses medium by DNA microarray analysis. (United States)

    Shima, Jun; Kuwazaki, Seigo; Tanaka, Fumiko; Watanabe, Hajime; Yamamoto, Hideki; Nakajima, Ryoichi; Tokashiki, Tadaaki; Tamura, Hiromi


    Genes whose expression levels are enhanced or reduced during the cultivation process that uses cane molasses in baker's yeast production were identified in this study. The results showed that baker's yeast grown in molasses medium had higher fermentation ability and stress tolerance compared with baker's yeast grown in synthetic medium. Molasses apparently provided not only sugar as a carbon source but also provided functional components that enhanced or reduced expression of genes involved in fermentation ability and stress tolerance. To identify the genes whose expression is enhanced or reduced during cultivation in molasses medium, DNA microarray analysis was then used to compare the gene expression profile of cells grown in molasses with that of cells grown in synthetic medium. To simulate the commercial baker's yeast production process, cells were cultivated using a fed-batch culture system. In molasses medium, genes involved in the synthesis or uptake of vitamins (e.g., biotin, pyridoxine and thiamine) showed enhanced expression, suggesting that vitamin concentrations in molasses medium were lower than those in synthetic medium. Genes involved in formate dehydrogenase and maltose assimilation showed enhanced expression in molasses medium. In contrast, genes involved in iron utilization (e.g., siderophore, iron transporter and ferroxidase) showed enhanced expression in synthetic medium, suggesting that iron starvation occurred. The genes involved in the metabolism of amino acids also showed enhanced expression in synthetic medium. This identification of genes provides information that will help improve the baker's yeast production process.

  5. Industrial textile effluent decolourization in stirred and static batch cultures of a new fungal strain Chaetomium globosum IMA1 KJ472923. (United States)

    Manai, Imène; Miladi, Baligh; El Mselmi, Abdellatif; Smaali, Issam; Ben Hassen, Aida; Hamdi, Moktar; Bouallagui, Hassib


    The treatment of an industrial textile effluent (ITE) was investigated by using a mono-culture of a novel fungal strain Chaetomium globosum IMA1. This filamentous fungus was selected based on its capacity for dye removal via the biodegradation mechanism. The respirometric analysis showed that C. globosum IMA1 was resistant to an indigo concentration up to 700 mg equivalent COD/L. The decolourization of the ITE by C. globosum was performed in static and stirred batch systems. The better lignin peroxidase (LiP), laccase and the manganese peroxidase (MnP) productions were 829.9 U/L, 83 U/L and 247.8 U/L, respectively since 3-5 days under a stirred condition. Therefore, the chemical oxygen demand (COD) and colors (OD620) removal yields reached 88.4% and 99.8%, respectively. Fourier transforms infrared spectroscopy (FTIR) analysis of the treated effluent showed that the decolourization was due to the degradation and the transformation of dye molecules. However, spectrophotometric examination showed that the complete dye removal was through fungal adsorption (8%), followed by degradation (92%).

  6. National culture and cultural orientations of owners affecting the innovation-growth relationship in five countries

    NARCIS (Netherlands)

    Rauch, Andreas; Frese, Michael; Wang, Zhong-Ming; Unger, Jens; Lozada, Maria; Kupcha, Vita; Spirina, Tanja


    This study tests the cross-cultural validity of the relationship between innovation and growth in a sample of 857 business owners from five different countries: China, Germany, the Netherlands, Peru and Russia. We found that innovation is effective in each country, suggesting universal relationships

  7. Polyamines in relation to growth in carrot cell cultures. (United States)

    Fallon, K M; Phillips, R


    Changes in polyamine metabolism were investigated in relation to growth of cell suspension cultures of carrot (Daucus carota, cv Chantenay). Changes in levels of the major amines putrescine and spermidine throughout the culture period correlated poorly with changes in fresh weight, but a closer correlation with the minor component spermine was observed. The arginine decarboxylase (ADC) inhibitor difluoromethylarginine (DFMA) strongly and specifically inhibited ADC activity in the supernatant, reduced the major amine (putrescine) by 95% and the total amine content by 80%. It had no effect on cell number and stimulated fresh weight by over 25% through increased cell expansion. Spermine content, in contrast, increased with DFMA concentration in parallel with fresh weight increases. Difluoromethylornithine strongly inhibited ornithine decarboxylase activity in the pellet, but had little effect on either polyamine levels or culture growth. It was concluded that little evidence for a correlation between free polyamines and cell number in carrot cultures could be detected, but that a possible correlation between spermine content and cell expansion was observed.

  8. Polyamines in Relation to Growth in Carrot Cell Cultures 1 (United States)

    Fallon, Kevin M.; Phillips, Richard


    Changes in polyamine metabolism were investigated in relation to growth of cell suspension cultures of carrot (Daucus carota, cv Chantenay). Changes in levels of the major amines putrescine and spermidine throughout the culture period correlated poorly with changes in fresh weight, but a closer correlation with the minor component spermine was observed. The arginine decarboxylase (ADC) inhibitor difluoromethylarginine (DFMA) strongly and specifically inhibited ADC activity in the supernatant, reduced the major amine (putrescine) by 95% and the total amine content by 80%. It had no effect on cell number and stimulated fresh weight by over 25% through increased cell expansion. Spermine content, in contrast, increased with DFMA concentration in parallel with fresh weight increases. Difluoromethylornithine strongly inhibited ornithine decarboxylase activity in the pellet, but had little effect on either polyamine levels or culture growth. It was concluded that little evidence for a correlation between free polyamines and cell number in carrot cultures could be detected, but that a possible correlation between spermine content and cell expansion was observed. PMID:16666271

  9. Modelling the growth kinetics of Kocuria marina DAGII as a function of single and binary substrate during batch production of β-Cryptoxanthin. (United States)

    Mitra, Ruchira; Chaudhuri, Surabhi; Dutta, Debjani


    In the present investigation, growth kinetics of Kocuria marina DAGII during batch production of β-Cryptoxanthin (β-CRX) was studied by considering the effect of glucose and maltose as a single and binary substrate. The importance of mixed substrate over single substrate has been emphasised in the present study. Different mathematical models namely, the Logistic model for cell growth, the Logistic mass balance equation for substrate consumption and the Luedeking-Piret model for β-CRX production were successfully implemented. Model-based analyses for the single substrate experiments suggested that the concentrations of glucose and maltose higher than 7.5 and 10.0 g/L, respectively, inhibited the growth and β-CRX production by K. marina DAGII. The Han and Levenspiel model and the Luong product inhibition model accurately described the cell growth in glucose and maltose substrate systems with a R (2) value of 0.9989 and 0.9998, respectively. The effect of glucose and maltose as binary substrate was further investigated. The binary substrate kinetics was well described using the sum-kinetics with interaction parameters model. The results of production kinetics revealed that the presence of binary substrate in the cultivation medium increased the biomass and β-CRX yield significantly. This study is a first time detailed investigation on kinetic behaviours of K. marina DAGII during β-CRX production. The parameters obtained in the study might be helpful for developing strategies for commercial production of β-CRX by K. marina DAGII.

  10. Batch and Fed-Batch Fermentation System on Ethanol Production from Whey using Kluyveromyces marxianus

    Directory of Open Access Journals (Sweden)

    H Hadiyanto


    Full Text Available Nowadays reserve of fossil fuel has gradually depleted. This condition forces many researchers to  find energy alternatives which is renewable and sustainable in the future. Ethanol derived from cheese industrial waste (whey using fermentation process can be a new perspective in order to secure both energy and environment. The aim of this study was  to compare the operation modes (batch and fed-batch of fermentation system on ethanol production from whey using Kluyveromyces marxianus. The result showed that the fermentation process for ethanol production by fed-batch system was higher at some point of parameters compared with batch system. Growth rate and ethanol yield (YP/S of fed-batch fermentation were 0.122/h and 0.21 gP/gS respectively; growth rate and ethanol yield (YP/S of batch fermentation were 0.107/h, and 0.12 g ethanol/g substrate, respectively. Based on the data of biomass and ethanol concentrations, the fermentation process for ethanol production by fed-batch system were higher at some point of parameters compared to batch system. Periodic substrate addition performed on fed-batch system leads the yeast growth in low substrate concentrations and consequently  increasing their activity and ethanol productivity. Keywords: batch; ethanol; fed-batch; fermentation;Kluyveromyces marxianus, whey

  11. Fed-batch pediocin production by Pediococcus acidilactici NRRL B-5627 on whey. (United States)

    Pérez Guerra, Nelson; Bernárdez, Paula Fajardo; Agrasar, Ana Torrado; López Macías, Cristina; Castro, Lorenzo Pastrana


    Cell growth and pediocin production by Pediococcus acidilactici NRRL B-5627 on whey were compared by using batch fermentation and re-alkalized fed-batch fermentation. The batch fermentations were performed on DWG [DW (diluted whey) supplemented with 1% (w/v) glucose], DWYE [DW supplemented with 2% (w/v) yeast extract] and DWGYE (DW supplemented with 1% glucose plus 2% yeast extract) media. The fed-batch culture on DWYE medium was fed with a mixture of concentrated whey (48 g of total sugars/l) supplemented with 2% yeast extract and 400 g/l concentrated glucose. The re-alkalized fed-batch culture was characterized by higher biomass (6.57 g/l) and pediocin [517.6 BU (bacteriocin activity units)/ml] concentrations compared with the batch processes on MRS (de Man, Rogosa and Sharpe) broth (1.76 g/l and 493.2 BU/ml), DW (0.17 g/l and 57.7 BU/ml), DWG (0.14 g/l and 53.6 BU/ml), DWYE (1.43 g/l and 187.6 BU/ml) and DWGYE (1.28 g/l and 167.3 BU/ml) media. A mixed acid fermentation was observed during the growth of P. acidilactici NRRL B-5627 in the fed-batch culture on DWYE medium, and other products (acetic acid and ethanol) in addition to lactic acid accumulated in the medium. Mathematical models were set up to describe fed-batch production of biomass and pediocin by P. acidilactici. The models developed offer a better fit and a more realistic description of the experimental biomass and pediocin production data when compared with the logistic and Luedeking and Piret model.

  12. Influence of extracellular pH on growth, viability, cell size, acidification activity, and intracellular pH of Lactococcus lactis in batch fermentations. (United States)

    Hansen, Gunda; Johansen, Claus Lindvald; Marten, Gunvor; Wilmes, Jacqueline; Jespersen, Lene; Arneborg, Nils


    In this study, we investigated the influence of three extracellular pH (pHex) values (i.e., 5.5, 6.5, and 7.5) on the growth, viability, cell size, acidification activity in milk, and intracellular pH (pHi) of Lactococcus lactis subsp. lactis DGCC1212 during pH-controlled batch fermentations. A universal parameter (e.g., linked to pHi) for the description or prediction of viability, specific acidification activity, or growth behavior at a given pHex was not identified. We found viability as determined by flow cytometry to remain high during all growth phases and irrespectively of the pH set point. Furthermore, regardless of the pHex, the acidification activity per cell decreased over time which seemed to be linked to cell shrinkage. Flow cytometric pHi determination demonstrated an increase of the averaged pHi level for higher pH set points, while the pH gradient (pHi-pHex) and the extent of pHi heterogeneity decreased. Cells maintained positive pH gradients at a low pHex of 5.5 and even during substrate limitation at the more widely used pHex 6.5. Moreover, the strain proved able to grow despite small negative or even absent pH gradients at a high pHex of 7.5. The larger pHi heterogeneity at pHex 5.5 and 6.5 was associated with more stressful conditions resulting, e.g., from higher concentrations of non-dissociated lactic acid, while the low pHi heterogeneity at pHex 7.5 most probably corresponded to lower concentrations of non-dissociated lactic acid which facilitated the cells to reach the highest maximum active cell counts of the three pH set points.

  13. Growth Culture Conditions and Nutrient Signaling Modulating Yeast Chronological Longevity

    Directory of Open Access Journals (Sweden)

    Júlia Santos


    Full Text Available The manipulation of nutrient-signaling pathways in yeast has uncovered the impact of environmental growth conditions in longevity. Studies using calorie restriction show that reducing glucose concentration of the culture media is sufficient to increase replicative and chronological lifespan (CLS. Other components of the culture media and factors such as the products of fermentation have also been implicated in the regulation of CLS. Acidification of the culture media mainly due to acetic acid and other organic acids production negatively impacts CLS. Ethanol is another fermentative metabolite capable of inducing CLS reduction in aged cells by yet unknown mechanisms. Recently, ammonium was reported to induce cell death associated with shortening of CLS. This effect is correlated to the concentration of NH4+ added to the culture medium and is particularly evident in cells starved for auxotrophy-complementing amino acids. Studies on the nutrient-signaling pathways regulating yeast aging had a significant impact on aging-related research, providing key insights into mechanisms that modulate aging and establishing the yeast as a powerful system to extend knowledge on longevity regulation in multicellular organisms.

  14. Fed-batch bioreactor process with recombinant Saccharomyces cerevisiae growing on cheese whey

    Directory of Open Access Journals (Sweden)

    R. Rech


    Full Text Available Saccharomyces cerevisiae strain W303 was transformed with two yeast integrative plasmids containing Kluyveromyces lactis LAC4 and LAC12 genes that codify beta-galactosidase and lactose permease respectively. The BLR030 recombinant strain was selected due to its growth and beta-galactosidase production capacity. Different culture media based on deproteinized cheese whey (DCW were tested and the best composition (containing DCW, supplemented with yeast extract 1 %, and peptone 3 % (w/v was chosen for bioreactor experiments. Batch, and fed-batch cultures with linear ascending feeding for 25 (FB25, 35 (FB35, and 50 (FB50 hours, were performed. FB35 and FB50 produced the highest beta-galactosidase specific activities (around 1,800 U/g cells, and also the best productivities (180 U/L.h. Results show the potential use of fed-batch cultures of recombinant S. cerevisiae on industrial applications using supplemented whey as substrate.

  15. Pro Spring Batch

    CERN Document Server

    Minella, Michael T


    Since its release, Spring Framework has transformed virtually every aspect of Java development including web applications, security, aspect-oriented programming, persistence, and messaging. Spring Batch, one of its newer additions, now brings the same familiar Spring idioms to batch processing. Spring Batch addresses the needs of any batch process, from the complex calculations performed in the biggest financial institutions to simple data migrations that occur with many software development projects. Pro Spring Batch is intended to answer three questions: *What? What is batch processing? What

  16. Increasing cell culture population doublings for long-term growth of finite life span human cell cultures (United States)

    Stampfer, Martha R; Garbe, James C


    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Stampfer, Martha R.; Garbe, James C.


    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.

  18. Comparison of transcriptional heterogeneity of eight genes between batch Desulfovibrio vulgaris biofilm and planktonic culture at a single-cell level

    Directory of Open Access Journals (Sweden)

    Zhenhua eQi


    Full Text Available Sulfate-reducing bacteria (SRB biofilm formed on metal surfaces can change the physicochemical properties of metals and cause metal corrosion. To enhance understanding of differential gene expression in Desulfovibrio vulgaris under planktonic and biofilm growth modes, a single-cell based RT-qPCR approach was applied to determine gene expression levels of 8 selected target genes in four sets of the 31 individual cells isolated from each growth condition (i.e., biofilm formed on a stainless steel (SS) and planktonic cultures, exponential and stationary phases. The results showed obvious gene-expression heterogeneity for the target genes among D. vulgaris single cells of both biofilm and planktonic cultures. In addition, an increased gene-expression heterogeneity in the D. vulgaris biofilm when compared with the planktonic culture was also observed for seven out of eight selected genes, which may be contributing to the increased complexity in terms of structures and morphology in the biofilm. Moreover, the results showed up-regulation of DVU0281 gene encoding exopolysaccharide biosynthesis protein, and down-regulation of genes involved in energy metabolism (i.e., DVU0434 and DVU0588, stress responses (i.e., DVU2410 and response regulator (i.e., DVU3062 in the D. vulgaris biofilm cells. Finally, the gene (DVU2571 involved in iron transportation was found down-regulated, and two genes (DVU1340 and DVU1397 involved in ferric uptake repressor and iron storage were up-regulated in D. vulgaris biofilm, suggesting their possible roles in maintaining normal metabolism of the D. vulgaris biofilm under environments of high concentration of iron. This study showed that the single-cell based analysis could be a useful approach in deciphering metabolism of microbial biofilms.

  19. [Peculiarities of growth and development of cultured mucosal cells from the upper respiratory tract stimulated by growth factors]. (United States)

    Chekan, V L; Kvacheva, Z B; Petrova, L G


    Specific features of growth and development of cultured mucosal cells from the upper respiratory tract were studied during their in vitro stimulation by keratinocyte growth factor (KGF) and epidermal growth factor (EGF). Phenotypic composition and quantitative characteristics of cultured epithelial cells was investigated with the use of monoclonal CD49F antibodies and flow cytofluorometry. The culture technique makes it possible to obtain a large amount of cells for the evaluation of their pathological changes. Moreover, cell cultures can be used to restore lesioned mucosa of the upper respiratory tract both in experiment and under clinical conditions.

  20. Cultivo mixotrófico da microalga Spirulina platensis em batelada alimentada Mixotrophic growth of Spirulina platensis in fed-batch mode

    Directory of Open Access Journals (Sweden)

    Adriana Muliterno


    Full Text Available A Spirulina platensis tem sido estudada devido a seu alto valor protéico, digestibilidade e por apresentar quantidades significativas de ácidos graxos poliinsaturados, vitaminas, fenólicos e ficocianina, podendo ser utilizada na alimentação humana. A utilização de nutrientes de baixo custo é um fator importante na produção da cianobactéria por possibilitar a redução de custos de processo. Objetivou-se com este trabalho estudar o cultivo mixotrófico da S. platensis por meio da adição de uma fonte orgânica de carbono (glicose em modo bateladaalimentada. Foi utilizado um Planejamento Fatorial Completo 2³ para o cultivo e as variáveis de estudo foram a concentração de glicose (0,5 gL-1 e 1,0 gL-1, a diluição do meio Zarrouk (50% e 75% e a iluminância (1800 lux e 3000 lux. A concentração celular máxima obtida foi de 5,38 gL-1 com uma velocidade específica máxima de crescimento de 0,0063 h-1, nas condições de 0,5 gL-1 de glicose, diluição do meio de 75% e iluminância de 3000 lux.The cyanobacterium Spirulina platensis has been studied due to its high content (~65% of highly digestible protein as well as significant amounts of polyunsaturated fatty acids, phenolics, vitamins, minerals and phycocyanin which could be useful in the human nutrition. The use of nutrients of low costs in the cyanobacterium growth could reduce the costs of production. We studied the fed-batch mixotrophic growth of the S. platensis in Zarrouk's medium with glucose (0.5 gL-1 and 1.0 gL-1 as carbon source and also investigated the effects of dilution (50% and 75%, with water and illumination (1,800 lux and 3,000 lux using a 2³ factorial design. The maximum celular concentration of 5.38 gL-1 and maximum specific growth rate of 0.0063 h-1 were obtained with a glucose concentration of 0.5 gL-1, 50% dilution and 1800 lux of illuminance.

  1. In vitro zygotic embryo culture of Pinus peuce Gris.: Optimization of culture conditions affecting germination and early seedling growth

    Directory of Open Access Journals (Sweden)

    Stojičić Dragana


    Full Text Available This study reports a protocol for the germination and early seedling growth of Pinus peuce Gris. using zygotic embryo culture. In order to overcome seed dormancy and optimize organogenesis, the effect of nutritional, plant growth regulatory and physical factors on in vitro germination and growth of isolated mature zygotic embryos of P. peuce were investigated.

  2. Estimation of autotrophic maximum specific growth rate constant--experience from the long-term operation of a laboratory-scale sequencing batch reactor system. (United States)

    Su, Yu-min; Makinia, Jacek; Pagilla, Krishna R


    The autotrophic maximum specific growth rate constant, muA,max, is the critical parameter for design and performance of nitrifying activated sludge systems. In literature reviews (i.e., Henze et al., 1987; Metcalf and Eddy, 1991), a wide range of muA,max values have been reported (0.25 to 3.0 days(-1)); however, recent data from several wastewater treatment plants across North America revealed that the estimated muA,max values remained in the narrow range 0.85 to 1.05 days(-1). In this study, long-term operation of a laboratory-scale sequencing batch reactor system was investigated for estimating this coefficient according to the low food-to-microorganism ratio bioassay and simulation methods, as recommended in the Water Environment Research Foundation (Alexandria, Virginia) report (Melcer et al., 2003). The estimated muA,max values using steady-state model calculations for four operating periods ranged from 0.83 to 0.99 day(-1). The International Water Association (London, United Kingdom) Activated Sludge Model No. 1 (ASM1) dynamic model simulations revealed that a single value of muA,max (1.2 days(-1)) could be used, despite variations in the measured specific nitrification rates. However, the average muA,max was gradually decreasing during the activated sludge chlorination tests, until it reached the value of 0.48 day(-1) at the dose of 5 mg chlorine/(g mixed liquor suspended solids x d). Significant discrepancies between the predicted XA/YA ratios were observed. In some cases, the ASM1 predictions were approximately two times higher than the steady-state model predictions. This implies that estimating this ratio from a complex activated sludge model and using it in simple steady-state model calculations should be accepted with great caution and requires further investigation.

  3. Growth response and toxin concentration of cultured Pyrodinium bahamense var. compressum to varying salinity and temperature conditions. (United States)

    Gedaria, Alice Ilaya; Luckas, Bernd; Reinhardt, Katrin; Azanza, Rhodora V


    The growth and toxin production of a Philippine Pyrodinium bahamense isolate in nutrient replete batch cultures were investigated under conditions affected by varying salinity, temperature and combined effects of salinity and temperature. Early exponential growth stage was reached after 7 days with a cell division rate of 0.26 div day(-1). The toxin content reached a peak of 298 fmol cell-1 at mid exponential phase and rapidly declined to 54 fmol cell-1 as it approached the death phase. Only three sets of toxins composed of STX, dcSTX and B1 were detected in which STX made up to 85-98 mol%toxincell-1. P. bahamense was able to grow in salinities and temperatures ranging from 26 per thousand to 36 per thousand and 23 to 36 degrees C, respectively. The optimum growth under varying salinity and temperature conditions was observed at 36 per thousand and 25 degrees C. Toxin content reached a peak of 376 fmol cell-1 at 25 degrees C and was lower (80-116 fmol cell-1) at higher temperatures (32-35 degrees C). Combined effects of salinity and temperature showed that P. bahamense was not able to grow at low salinity and temperature (i.e. below 26 per thousand-28 degrees C). Optimum growth was observed in higher salinities at all temperature conditions.

  4. Improved elastase production by Bacillus sp. EL31410—further optimization and kinetics studies of culture medium for batch fermentation

    Institute of Scientific and Technical Information of China (English)

    何国庆; 陈启和; 琚晓捷; 石乃冬


    An efficient culture medium producing a bacterial elastase with high yields was developed further following preliminary studies by means of response surface method. Central composite design (CCD) and response surface methodology were applied to optimize the medium constituents. A central composite design was used to explain the combined effect of three medium constituents, viz, glucose, K2HPO4, MgSO4·7H2O. The strain produced more elastase in the completely optimized medium, as compared with the partially optimized medium. The fitted model of the second model, as per RSM, showed that glucose was 7.4 g/100 ml, casein 1.13 g/100 ml, corn steep flour 0.616 g/100 ml, KEHPO4 0.206 g/100 ml and MgSO4·7H2O 0.034 g/100 ml. The fermentation kinetics of these two culture media in the flask experiments were analyzed. It was found that the highest elastase productivity occurred at 54 hours. Higher glucose concentration had inhibitory effect on elastase production. At the same time, we observed that the glucose consumption rate was slow in the completely optimized medium, which can explain the lag period of the highest elastase production. Some metal ions and surfactant additives also affected elastase production and cell growth.

  5. 红曲霉色素流加培养的初步研究%Preliminary Study on Fed-batch Culture of Monascus Pigments

    Institute of Scientific and Technical Information of China (English)

    杨旭; 曹岚; 李旭


    Objective:The optimization methods of monascus pigments with submerged fermentation are studied.The level of fermentation is improved and the products contain a high concentration of monascus pigments.Methods:L8 (27) orthogonal experiment and fed-batch fermentation method of monaseus purpureus liquid are designed to select the most efficient fermentation medium for monaseus purpureus,reduce inhibition caused by over-rich nutrients and improve the fermentation concentration and level through different batch fermentation modes.Conclusion:Optimized shake flask fermentation medium with maltose of 8°Bx,soluble starch of 3%,soya bean protein powder of 4%,NaCl of 0.5%,magnesium sulfate of 0.05% and dipotassium hydrogen phosphate of 0.1%.The optimum fed-batch mode is obtained by fed-batch fermentation for 2 L batch tank,the best feeding submerged fermentation starts feeding after 60 h,filling every 20 min,completes after 12 h.Compared with batch fermentation,the valence of fed-batch fermented monascus pigment increases by 57%.%目的:研究红曲霉液体深层发酵的优化方法,提高其发酵水平,生产出含有较高红曲霉色素的产品.方法:采用L8(27)正交实验法和红曲霉液体流加发酵方法,通过不同的补料发酵方式,降低营养物质过浓而产生的阻碍作用,提高发酵浓度和水平.结果:优化后的培养基为饴糖8°Bx,可溶淀粉3%,大豆蛋白粉4%,氯化钠0.5%,硫酸镁0.05%,磷酸氢二钾0.1%.通过对2L发酵罐的补料发酵得出最佳的补料方式:液体深层发酵60 h以后开始补料,每20 min补1次,12 h补完,同分批发酵相比,流加发酵红曲色素的效价提高57%.

  6. Growth and biomass production with enhanced {beta}-glucan and dietary fibre contents of Ganoderma australe ATHUM 4345 in a batch-stirred tank bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Papaspyridi, Lefki-Maria; Christakopoulos, Paul [BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, Athens (Greece); Katapodis, Petros [BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, Athens (Greece); Biotechnology Laboratory, Department of Biological Applications and Technologies, University of Ioannina, Ioannina (Greece); Gonou-Zagou, Zacharoula; Kapsanaki-Gotsi, Evangelia [Department of Ecology and Systematics, Faculty of Biology, National and Kapodistrian University of Athens, Athens (Greece)


    In this study we maximized biomass production by the basidiomycete Ganoderma australe ATHUM 4345, a species of pharmaceutical interest as it is a valuable source of nutraceuticals, including dietary fibers and glucans. We used the Biolog FF MicroPlate to screen 95 different carbon sources for growth monitoring. The pattern of substrate catabolism forms a substrate assimilation fingerprint, which is useful in selecting components for media optimization of maximum biomass production. Response surface methodology, based on the central composite design was applied to explore the optimum concentrations of carbon and nitrogen sources of culture medium in shake flask cultures. When the improved culture medium was tested in a 20-L stirred tank bioreactor, using 13.7 g/L glucose and 30.0 g/L yeast extract, high biomass yields (10.1{+-}0.4 g/L) and productivity of 0.09 g L{sup -1} h{sup -1} were obtained. The yield coefficients for total glucan and dietary fibers on biomass formed were 94.82{+-}6 and 341.15{+-}12.3 mg/g mycelium dry weight, respectively. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  7. Growth detection failures by the nonradiometric Bactec MGIT 960 mycobacterial culture system. (United States)

    Peña, Jeremy A; Ferraro, Mary Jane; Hoffman, Colleen G; Branda, John A


    Mycobacterial growth in liquid culture can go undetected by automated, nonradiometric growth detection systems. In our laboratory, instrument-negative tubes from the Bactec MGIT 960 system are inspected visually for clumps suggestive of mycobacterial growth, which (if present) are examined by acid-fast smear analysis. A 3-year review demonstrated that ∼1% of instrument-negative MGIT cultures contained mycobacterial growth and that 10% of all cultures yielding mycobacteria were instrument negative. Isolates from instrument-negative MGIT cultures included both tuberculous and nontuberculous mycobacteria.

  8. Prediction of competitive microbial growth in mixed culture at dynamic temperature patterns. (United States)

    Fujikawa, Hiroshi; Sakha, Mohammad Z


    A novel competition model developed with the new logistic model and the Lotka-Volterra model successfully predicted the growth of bacteria in mixed culture using the mesophiles Staphylococcus aureus, Escherichia coli, and Salmonella at a constant temperature in our previous studies. In this study, we further studied the prediction of the growth of those bacteria in mixed culture at dynamic temperatures with various initial populations with the competition model. First, we studied the growth kinetics of the species in a monoculture at various constant temperatures ranging from 16℃ to 32℃. With the analyzed data in the monoculture, we then examined the prediction of bacterial growth in mixed culture with two and three species. The growth of the bacteria in the mixed culture at dynamic temperatures was successfully predicted with the model. The residuals between the observed and predicted populations at the data points were growth in mixed culture at dynamic temperature patterns.

  9. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

    Directory of Open Access Journals (Sweden)

    Sutini Sutini


    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  10. Spring batch essentials

    CERN Document Server

    Rao, P Raja Malleswara


    If you are a Java developer with basic knowledge of Spring and some experience in the development of enterprise applications, and want to learn about batch application development in detail, then this book is ideal for you. This book will be perfect as your next step towards building simple yet powerful batch applications on a Java-based platform.

  11. Growth of the Pittsburgh Pneumonia Agent in Animal Cell Cultures


    Rinaldo, Charles R.; Pasculle, A. William; Myerowitz, Richard L.; Gress, Francis M.; Dowling, John N.


    Pittsburgh pneumonia agent (Legionella micdadei) grew in monkey, chicken, and human cell cultures. Pittsburgh pneumonia agent grew predominantly in the cytoplasm, resulting in a nonfocal, mild cytopathic effect.

  12. Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis. (United States)

    Peretz, Avi; Geffen, Yuval; Socea, Soergiu D; Pastukh, Nina; Graffi, Shmuel


    Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media.

  13. Growth of cultured porcine retinal pigment epithelial cells

    DEFF Research Database (Denmark)

    Wiencke, A.K.; Kiilgaard, Jens Folke; Nicolini, Jair;


    To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation.......To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation....

  14. 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.)


    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.

  15. Involvement of nucleotides in glial growth following scratch injury in avian retinal cell monolayer cultures. (United States)

    Silva, Thayane Martins; França, Guilherme Rapozeiro; Ornelas, Isis Moraes; Loiola, Erick Correia; Ulrich, Henning; Ventura, Ana Lucia Marques


    When retinal cell cultures were mechanically scratched, cell growth over the empty area was observed. Only dividing and migrating, 2 M6-positive glial cells were detected. Incubation of cultures with apyrase (APY), suramin, or Reactive Blue 2 (RB-2), but not MRS 2179, significantly attenuated the growth of glial cells, suggesting that nucleotide receptors other than P2Y1 are involved in the growth of glial cells. UTPγS but not ADPβS antagonized apyrase-induced growth inhibition in scratched cultures, suggesting the participation of UTP-sensitive receptors. No decrease in proliferating cell nuclear antigen (PCNA(+)) cells was observed at the border of the scratch in apyrase-treated cultures, suggesting that glial proliferation was not affected. In apyrase-treated cultures, glial cytoplasm protrusions were smaller and unstable. Actin filaments were less organized and alfa-tubulin-labeled microtubules were mainly parallel to scratch. In contrast to control cultures, very few vinculin-labeled adhesion sites could be noticed in these cultures. Increased Akt and ERK phosphorylation was observed in UTP-treated cultures, effect that was inhibited by SRC inhibitor 1 and PI3K blocker LY294002. These inhibitors and the FAK inhibitor PF573228 also decreased glial growth over the scratch, suggesting participation of SRC, PI3K, and FAK in UTP-induced growth of glial cells in scratched cultures. RB-2 decreased dissociated glial cell attachment to fibronectin-coated dishes and migration through transwell membranes, suggesting that nucleotides regulated adhesion and migration of glial cells. In conclusion, mechanical scratch of retinal cell cultures induces growth of glial cells over the empty area through a mechanism that is dependent on activation of UTP-sensitive receptors, SRC, PI3K, and FAK.

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

    Directory of Open Access Journals (Sweden)

    Villegas E Valeska


    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.


    Hershey, A D; Bronfenbrenner, J


    1. Measurements of the rate of oxygen uptake per cell in transplants of Bacterium coli from cultures of this organism in different phases of growth have given results in essential agreement with the observations of others. 2. Correlations of viable count, centrifugable nitrogen, and turbidity, with oxygen consumption, indicate that the increased metabolism during the early portion of the growth period is quantitatively referable to increased average size of cells. 3. Indirect evidence has suggested that the initial rate of growth of transplants is not related to the phase of growth of the parent culture.

  18. Biosynthesis of highly enriched 13C-lycopene for human metabolic studies using repeated batch tomato cell culturing with 13C-glucose


    Moran, Nancy E.; Rogers, Randy B.; Lu, Chi-Hua; Conlon, Lauren E.; Lila, Mary Ann; Clinton, Steven K.; Erdman, John W


    While putative disease-preventing lycopene metabolites are found in both tomato (Solanum lycopersicum) products and in their consumers, mammalian lycopene metabolism is poorly understood. Advances in tomato cell culturing techniques offer an economical tool for generation of highly-enriched 13C-lycopene for human bioavailability and metabolism studies. To enhance the 13C-enrichment and yields of labeled lycopene from the hp-1 tomato cell line, cultures were first grown in 13C-glucose media fo...

  19. Product formation from thiophene by a mixed bacterial culture. Influence of benzene as growth substrate

    DEFF Research Database (Denmark)

    Rivas, Isabelle Marie; Mosbæk, Hans; Arvin, Erik


    The influence of benzene as a growth substrate on the cometabolic conversion of thiophene was investigated in batch systems with microorganisms originating from an creosote contaminated site. Benzene was shown to stimulate the conversion of thiophene with a first-order rate, during the initial...... phase of transformation. The microorganisms were able to transform thiophene in the absence of benzene at a zero-order rate. Thiophene was converted to five oxidation products, regardless of the presence of benzene. Benzene had no influence on the distribution of these oxidation products. The main...

  20. Building a Culture of Growth and Evaluation in Schools (United States)

    Hall, Catherine J.


    This author contends that there are four key ingredients to a growth mindset in schools: observation, dialog, feedback, and investment. In addition, teachers and supervisors need to be jointly invested in the growth mindset of the school. The author suggests that if a school is considering a new faculty evaluation system, it is crucial that…

  1. 6-Gingerol inhibits hair shaft growth in cultured human hair follicles and modulates hair growth in mice.

    Directory of Open Access Journals (Sweden)

    Yong Miao

    Full Text Available Ginger (Zingiber officinale has been traditionally used to check hair loss and stimulate hair growth in East Asia. Several companies produce shampoo containing an extract of ginger claimed to have anti-hair loss and hair growth promotion properties. However, there is no scientific evidence to back up these claims. This study was undertaken to measure 6-gingerol, the main active component of ginger, on hair shaft elongation in vitro and hair growth in vivo, and to investigate its effect on human dermal papilla cells (DPCs in vivo and in vitro. 6-Gingerol suppressed hair growth in hair follicles in culture and the proliferation of cultured DPCs. The growth inhibition of DPCs by 6-gingerol in vitro may reflect a decrease in the Bcl-2/Bax ratio. Similar results were obtained in vivo. The results of this study showed that 6-gingerol does not have the ability to promote hair growth, on the contrary, can suppress human hair growth via its inhibitory and pro-apoptotic effects on DPCs in vitro, and can cause prolongation of telogen phase in vivo. Thus, 6-gingerol rather than being a hair growth stimulating drug, it is a potential hair growth suppressive drug; i.e. for hair removal.


    Institute of Scientific and Technical Information of China (English)


    Objective To ascertain whether the growth inhibitor in conditioned medium from cultured rabbit arte rial cells is distinct from TGF-β. Methods Rabbit aortic smooth muscle ceils were grown from explained segments of the aorta. Conditioned medium from cultured rabbit aortic smooth muscle ceils and anti-TGF-β were employed in this study. Smooth muscle cell proliferation was measured by XTT detection (Boehringer Mannheim). Results Acidified conditioned medium from smooth muscle ceils had significantly stronger effects of growth inhibition than controls, and anti-TGF-β did not affect the growth inhibitory effect of conditioned medium from cultured rabbit arterial smooth muscle cells. Conclusion The growth inhibiting substance in conditioned medium from cultured rabbit aortic smooth muscle cells is distinct from TGF-β.

  3. Kinetics of Growth and Nutrient Consumption in the Culture of Trichosanthes kirilowii Hairy Root

    Institute of Scientific and Technical Information of China (English)

    郭志刚; 郑明智; 刘瑞芝


    Ribosome-inactivating proteins in Trichosanthes kirilowii having high anti-HIV activity can be efficiently obtained by culturing Trichosanthes kirilowii hairy root. A hairy root line from Trichosanthes kirilowii was cultivated in flasks and in a 5-L bioreactor. The results show that a logistic equation can be used to describe the relationship between the hairy root biomass and the culture time. The medium conductivity decrease is linearly related to the biomass amount. In the suspension culture, the hairy root growth is closely related to the nutrient consumption. The biomass to nitrate yield is 0.352 g/mmol. The growth rate of the hairy root in the bioreactor is higher than that in the flasks, and after a 12-day culture its growth rate enters a high-speed period when the growth rate is 0.738 g/(L * d).

  4. The development from kinetic coefficients of a predictive model for the growth of Eichhomia crassipes in the field. I. Generating kinetic coefficients for the model in greenhouse culture

    Directory of Open Access Journals (Sweden)

    C. F. Musil


    Full Text Available The kinetics of N- and P- limited growth of Eichhornia crassipes (Mart . Solms were investigated in greenhouse culture with the object of developing a model for predicting population sizes, yields, growth rates and frequencies and amounts of harvest, under varying conditions of nutrient loading and climate, to control both nutrient inputs and excessive growth in eutrophied aquatic systems. The kinetic coefficients, maximum specific growth rate (Umax, half saturation coefficient (Ks and yield coefficient (Yc were measured under N and P limitation in replicated batch culture experiments. Umax values and Ks concentrations derived under N limitation ranged from 5,37 to 8,86% d + and from 400 to 1 506 µg  N ℓ1respectively. Those derived under P limitation ranged from 4,51 to 10,89% d 1 and from 41 to 162 fig P ℓ1 respectively. Yc values (fresh mass basis determined ranged from 1 660 to 1 981 (87 to 98 dry mass basis for N and from 16 431 to 18 671 (867 to 980 dry mass basis for P. The reciprocals of Yc values (dry mass basis, expressed as percentages, adequately estimated the minimum limiting concentrations of N and P {% dry mass in the plant tissues. Kinetic coefficients determined are compared with those reported for algae. The experimental method used and results obtained are critically assessed.

  5. Production of savinase and population viability of Bacillus clausii during high-cell-density fed-batch cultivations

    DEFF Research Database (Denmark)

    Christiansen, Torben; Michaelsen, S.; Wumpelmann, M.


    The growth and product formation of a Savinase-producing Bacillus clausii were investigated in high-cell-density fed-batch cultivations with both linear and exponential feed profiles. The highest specific productivity of Savinase was observed shortly after the end of the initial batch phase for all....... The physiological state of the cells was monitored during the cultivations using a flow cytometry assay based on the permeability of the cell membrane to propidium iodide. In the latter parts of the fed-batch cultures with a linear feed profile, a large portion of the cell population was found to have a permeable...... membrane, indicating a large percentage of dead cells. By assuming that only cells with a nonpermeable membrane contributed to growth and product formation, the physiological properties of this subpopulation were calculated....

  6. Small doses, big troubles: modeling growth dynamics of organisms affecting microalgal production cultures in closed photobioreactors. (United States)

    Forehead, Hugh I; O'Kelly, Charles J


    The destruction of mass cultures of microalgae by biological contamination of culture medium is a pervasive and expensive problem, in industry and research. A mathematical model has been formulated that attempts to explain contaminant growth dynamics in closed photobioreactors (PBRs). The model simulates an initial growth phase without PBR dilution, followed by a production phase in which culture is intermittently removed. Contaminants can be introduced at any of these stages. The model shows how exponential growth from low initial inocula can lead to "explosive" growth in the population of contaminants, appearing days to weeks after inoculation. Principal influences are contaminant growth rate, PBR dilution rate, and the size of initial contaminant inoculum. Predictions corresponded closely with observed behavior of two contaminants, Uronema sp. and Neoparamoeba sp., found in operating PBRs. A simple, cheap and effective protocol was developed for short-term prediction of contamination in PBRs, using microscopy and archived samples.

  7. Effects of different Helicobacter pylori culture filtrates on growth of gastric epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Yan-Guo Yan; Gang Zhao; Jin-Ping Ma; Shi-Rong Cai; Wen-Hua Zhan


    AIM: To study the effects of different Helicobacter pylori (H py/orl) culture filtrates on growth of gastric epithelial cells.METHODS: Broth culture filtrates of H pylori were prepared. Gastric epithelial cells were treated with the filtrates, and cell growth was determined by growth curve and flow cytometry. DNA damage of gastric epithelial cells was measured by single-cell microgel electrophoresis.RESULTS: Gastric epithelial cells proliferated actively when treated by CagA-gene-positive broth culture filtrates, and colony formation reached 40%. The number of cells in S phase increased compared to controls. Comet assay showed 41.2% comet cells in GES-1 cells treated with CagA-positive filtrates (P<0.05).CONCLUSION: CagA-positive filtrates enhance the changes in morphology and growth characteristics of human gastric epithelial tumor cells. DNA damage maybe one of the mechanisms involved in the growth changes.

  8. Growth and rutin production in hairy root cultures of buckwheat (Fagopyrum esculentum M.). (United States)

    Lee, Sook-Young; Cho, Soo-In; Park, Min-Hee; Kim, Yong-Kyung; Choi, Jae-Eul; Park, Sang-Un


    Buckwheat (Fagopyrum esculentum Moench.) is a potentially important source of rutin, a natural flavonoid with antihyperglycemic, antihypertensive, and antioxidative properties. To examine in vitro production of rutin, we established a hairy root culture of buckwheat by infecting leaf explants with Agrobacterium rhizogenes R1000, and tested the growth conditions and rutin production rates of these cultures. Ten hairy root clones were established; their growth and rutin production rates ranged from 233 to 312 (mg dry wt per 30 mL flask, and 0.8 to 1.2 (mg/g dry wt), respectively. Clone H8, which had high growth and rutin production rates (312 mg dry wt per 30 mL flask and 1.2 mg/g dry wt, respectively), was selected for further experiments. H8 showed maximal growth and rutin content at 30 days in culture in MS medium. Of four tested culture media, half-strength MS medium was found to induce the highest levels of growth (378 mg dry wt per 30 mL flask) and rutin production (1.4 mg/g dry wt) by clone H8. In contrast, supplementation with auxins (0.1-1 mg/l IAA, IBA and NAA) increased the growth rate, but had no significant effect on rutin production by H8. Collectively, these findings indicate that hairy root cultures of buckwheat culture could be a valuable alternative approach for rutin production.

  9. Fish Culture Data - Characterization of Sexual Growth Dimorphism in Sablefish (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Sexual growth dimorphism (SGD) is a common phenomenon in nature. Numerous marine fishes exhibit SGD, with females often growing faster and attaining larger sizes...

  10. Growth and Cultural Characteristics of Ophiocordyceps longissima Collected in Korea. (United States)

    Sung, Gi-Ho; Shrestha, Bhushan; Han, Sang-Kuk; Sung, Jae-Mo


    We investigated the effect of nutritional and environmental factors on Ophiocordyceps longissima mycelial growth. The longest colony diameter was observed on Schizophyllum (mushroom) genetics complete medium plus yeast extract, Schizophyllum (mushroom) genetics minimal medium, and Sabouraud dextrose agar (SDA); however, malt-extract yeast-extract agar, SDA plus yeast extract, yeast-extract malt-extract peptone dextrose agar, SDA, oatmeal agar, and potato dextrose agar showed higher mycelia density. A temperature of 25℃ was optimum and 7.0 was the optimum pH for mycelial growth. Colony diameter was similar under light and dark conditions. Maltose and yeast extract showed the highest mycelial growth among carbon and nitrogen sources respectively. The effect of mineral salts was less obvious; however, K(3)PO(4) showed slightly better growth than that of the other mineral salts tested. Among all nutrition sources tested, complex organic nitrogen sources such as yeast extract, peptone, and tryptone were best for mycelial growth of O. longissima. Ophiocordyceps longissima composite medium, formulated by adding maltose (2% w/v), yeast extract (1% w/v), and K(3)PO(4) (0.05% w/v) resulted in slightly longer colony diameter. In vitro mycelial O. longissima growth was sustainable and the production of fruiting bodies could be used for commercial purposes in the future.

  11. Hybrid Batch Bayesian Optimization

    CERN Document Server

    Azimi, Javad; Fern, Xiaoli


    Bayesian Optimization aims at optimizing an unknown non-convex/concave function that is costly to evaluate. We are interested in application scenarios where concurrent function evaluations are possible. Under such a setting, BO could choose to either sequentially evaluate the function, one input at a time and wait for the output of the function before making the next selection, or evaluate the function at a batch of multiple inputs at once. These two different settings are commonly referred to as the sequential and batch settings of Bayesian Optimization. In general, the sequential setting leads to better optimization performance as each function evaluation is selected with more information, whereas the batch setting has an advantage in terms of the total experimental time (the number of iterations). In this work, our goal is to combine the strength of both settings. Specifically, we systematically analyze Bayesian optimization using Gaussian process as the posterior estimator and provide a hybrid algorithm t...

  12. Cultural variation in early feeding pattern and maternal perceptions of infant growth

    NARCIS (Netherlands)

    van Eijsden, Manon; Meijers, Claire M. C.; Jansen, Jessica E.; De Kroon, Marlou L. A.; Vrijkotte, Tanja G. M.


    The perception of healthy growth and weight may differ between cultures, which could influence feeding practises and consequently affect the development of overweight. The present study examined ethnic variation in maternal perceptions of growth and their influence on feeding practises among Turkish

  13. Heuristics for batching and sequencing in batch processing machines

    Directory of Open Access Journals (Sweden)

    Chuda Basnet


    Full Text Available In this paper, we discuss the “batch processing” problem, where there are multiple jobs to be processed in flow shops. These jobs can however be formed into batches and the number of jobs in a batch is limited by the capacity of the processing machines to accommodate the jobs. The processing time required by a batch in a machine is determined by the greatest processing time of the jobs included in the batch. Thus, the batch processing problem is a mix of batching and sequencing – the jobs need to be grouped into distinct batches, the batches then need to be sequenced through the flow shop. We apply certain newly developed heuristics to the problem and present computational results. The contributions of this paper are deriving a lower bound, and the heuristics developed and tested in this paper.

  14. Effect of nitrogen salts on the growth of Ceratonia siliqua L. Shoot cultures

    Directory of Open Access Journals (Sweden)

    Vinterhalter Branka


    Full Text Available Effects of reduced nitrogen salt nutriton on the growth, lenticel hypertrophy and anthocyanin accumulation of carob (Ceratonia siliqua L. shoot cultures were investigated in conditions of light and darkness. Growth of shoot cultures was not significantly affected until nitrogen salts were reduced to less than ¼ of full-strength MS (Murashige and Skoog, 1962 values. Cultures in darkness were less affected and their main shoots even increased in length. Appearance of hypertrophied lenticels in light decreased, while in darkness they were absent in all treatments. Reduced nitrogen salt nutrition strongly affected anthocyanin accumulation of shoots and leaves, which greatly increased in both light and darkness. .

  15. Prediction of microbial growth in mixed culture with a competition model. (United States)

    Fujikawa, Hiroshi; Sakha, Mohammad Z


    Prediction of microbial growth in mixed culture was studied with a competition model that we had developed recently. The model, which is composed of the new logistic model and the Lotka-Volterra model, is shown to successfully describe the microbial growth of two species in mixed culture using Staphylococcus aureus, Escherichia coli, and Salmonella. With the parameter values of the model obtained from the experimental data on monoculture and mixed culture with two species, it then succeeded in predicting the simultaneous growth of the three species in mixed culture inoculated with various cell concentrations. To our knowledge, it is the first time for a prediction model for multiple (three) microbial species to be reported. The model, which is not built on any premise for specific microorganisms, may become a basic competition model for microorganisms in food and food materials.

  16. Nerve growth factor enhances DNA synthesis in cultured cerebellar neuroblasts. (United States)

    Confort, C; Charrasse, S; Clos, J


    The cerebellar neuroblasts in primary cultures from five-day-old rats bore NGF receptor immunoreactivity, suggesting a potential responsive to NGF. At low plating density, NGF was found to enhance DNA synthesis in these cells in a dose-dependent manner. As these cells synthesize NGF, one possibility to account for the lack of response of neuroblasts plated at high density is that the amount of endogenous trophic agent produced in this culture condition is sufficient to ensure an optimal effect. The results demonstrate that premitotic neuroblasts in the CNS, as well postmitotic neurons, are responsive to NGF. At the early stage of its development, the cerebellum therefore appears to be a very good autocrine model of NGF action.

  17. Growth and Cultural Characteristics of Cordyceps cardinalis Collected from Korea



    Cordyceps cardinalis was reported in Japan and the USA in 2004, and its fruiting bodies have recently been cultured in Korea. Herbarium specimens preserved at the Cordyceps Research Institute, Mushtech, Korea were revised and identified as C. cardinalis, based on morphological characters and conidial structures. Most of the C. cardinalis specimens were collected from Mt. Halla in Jeju-do. The effects of various nutritional sources and environmental conditions such as temperature and pH on myc...

  18. Quantitative modeling of viable cell density, cell size, intracellular conductivity, and membrane capacitance in batch and fed-batch CHO processes using dielectric spectroscopy. (United States)

    Opel, Cary F; Li, Jincai; Amanullah, Ashraf


    Dielectric spectroscopy was used to analyze typical batch and fed-batch CHO cell culture processes. Three methods of analysis (linear modeling, Cole-Cole modeling, and partial least squares regression), were used to correlate the spectroscopic data with routine biomass measurements [viable packed cell volume, viable cell concentration (VCC), cell size, and oxygen uptake rate (OUR)]. All three models predicted offline biomass measurements accurately during the growth phase of the cultures. However, during the stationary and decline phases of the cultures, the models decreased in accuracy to varying degrees. Offline cell radius measurements were unsuccessfully used to correct for the deviations from the linear model, indicating that physiological changes affecting permittivity were occurring. The beta-dispersion was analyzed using the Cole-Cole distribution parameters Deltaepsilon (magnitude of the permittivity drop), f(c) (critical frequency), and alpha (Cole-Cole parameter). Furthermore, the dielectric parameters static internal conductivity (sigma(i)) and membrane capacitance per area (C(m)) were calculated for the cultures. Finally, the relationship between permittivity, OUR, and VCC was examined, demonstrating how the definition of viability is critical when analyzing biomass online. The results indicate that the common assumptions of constant size and dielectric properties used in dielectric analysis are not always valid during later phases of cell culture processes. The findings also demonstrate that dielectric spectroscopy, while not a substitute for VCC, is a complementary measurement of viable biomass, providing useful auxiliary information about the physiological state of a culture.

  19. Glutamate enhances the expression of vascular endothelial growth factor in cultured SD rat astrocytes

    Institute of Scientific and Technical Information of China (English)


    Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups:control group (C),glutamate group (G),QA group (Q),DCG-IV group (D),L-AP4 group (L) and glutamate+MCPG group (G+M). Cells were cultured under nomoxic condition (95% air,5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes,respect...

  20. Development of a competition model for microbial growth in mixed culture. (United States)

    Fujikawa, Hiroshi; Munakata, Kanako; Sakha, Mohammad Z


    A novel competition model for describing bacterial growth in mixed culture was developed in this study. Several model candidates were made with our logistic growth model that precisely describes the growth of a monoculture of bacteria. These candidates were then evaluated for the usefulness in describing growth of two competing species in mixed culture using Staphylococcus aureus, Escherichia coli, and Salmonella. Bacterial cells of two species grew at initial doses of 10(3), 10(4), and 10(5) CFU/g at 28ºC. Among the candidates, a model where the Lotka-Volterra model, a general competition model in ecology, was incorporated as a new term in our growth model was the best for describing all types of growth of two competitors in mixed culture. Moreover, the values for the competition coefficient in the model were stable at various combinations of the initial populations of the species. The Baranyi model could also successfully describe the above types of growth in mixed culture when it was coupled with the Gimenez and Dalgaard model. However, the values for the competition coefficients in the competition model varied with the conditions. The present study suggested that our model could be a basic model for describing microbial competition.

  1. Oscillatory, stochastic and chaotic growth rate fluctuations in permittistatically controlled yeast cultures. (United States)

    Davey, H M; Davey, C L; Woodward, A M; Edmonds, A N; Lee, A W; Kell, D B


    We describe a continuous culture system related to the turbidostat, but using a feedback system based on biomass estimation from the dielectric permittivity of the cell suspension rather than its optical density. It is shown that this system provides an excellent method of maintaining a constant biomass level within a fermentor. The computer-controlled system was able to effect the essentially continuous registration of growth rate by monitoring the rate of medium addition via the time-dependent activity of the pump. At some biomass setpoints for aerobically grown cultures of baker's yeast substantial time-dependent fluctuations in the growth rate of the culture were thereby observed. At some biomass setpoints, however, or under anaerobic conditions, or when using a non-Crabtree yeast, the growth rate was constant, indicating that the fluctuations were inherent to the biological system and not simply a property of the fermentor and control system. A variety of time series analyses (Fourier transformations, Hurst and Lyapunov exponents, the determination of embedding dimension, and non-linear time series predictions based on the methodology of Sugihara and May) were used to demonstrate, for the first time, that as well as stochastic and periodic components these fluctuations exhibited deterministic chaos. 'Trivial predictors' were unable to give accurate predictions of the growth rate in these cultures. The growth rate fluctuations were studied further by means of offline measurements of changes in percentage viability, bud count, and in the external ethanol and glucose concentrations; these data and other evidence suggested that the growth rate fluctuations were closely linked to the primary respiro-fermentative metabolism of this organism. The identification of chaotic growth rates in cell cultures suggests that there may be novel methods for controlling the growth of such cultures.

  2. The fed-batch principle for the molecular biology lab: controlled nutrient diets in ready-made media improve production of recombinant proteins in Escherichia coli. (United States)

    Krause, Mirja; Neubauer, Antje; Neubauer, Peter


    While the nutrient limited fed-batch technology is the standard of the cultivation of microorganisms and production of heterologous proteins in industry, despite its advantages in view of metabolic control and high cell density growth, shaken batch cultures are still the standard for protein production and expression screening in molecular biology and biochemistry laboratories. This is due to the difficulty and expenses to apply a controlled continuous glucose feed to shaken cultures. New ready-made growth media, e.g. by biocatalytic release of glucose from a polymer, offer a simple solution for the application of the fed-batch principle in shaken plate and flask cultures. Their wider use has shown that the controlled diet not only provides a solution to obtain significantly higher cell yields, but also in many cases folding of the target protein is improved by the applied lower growth rates; i.e. final volumetric yields for the active protein can be a multiple of what is obtained in complex medium cultures. The combination of the conventional optimization approaches with new and easy applicable growth systems has revolutionized recombinant protein production in Escherichia coli in view of product yield, culture robustness as well as significantly increased cell densities. This technical development establishes the basis for successful miniaturization and parallelization which is now an important tool for synthetic biology and protein engineering approaches. This review provides an overview of the recent developments, results and applications of advanced growth systems which use a controlled glucose release as substrate supply.

  3. Growth and Cultural Characteristics of Ophiocordyceps longissima Collected in Korea



    We investigated the effect of nutritional and environmental factors on Ophiocordyceps longissima mycelial growth. The longest colony diameter was observed on Schizophyllum (mushroom) genetics complete medium plus yeast extract, Schizophyllum (mushroom) genetics minimal medium, and Sabouraud dextrose agar (SDA); however, malt-extract yeast-extract agar, SDA plus yeast extract, yeast-extract malt-extract peptone dextrose agar, SDA, oatmeal agar, and potato dextrose agar showed higher mycelia de...

  4. Growth arrest of vascular smooth muscle cells in suspension culture using low-acyl gellan gum. (United States)

    Natori, Tomomi; Fujiyoshi, Masachika; Uchida, Masashi; Abe, Natsuki; Kanaki, Tatsuro; Fukumoto, Yasunori; Ishii, Itsuko


    The proliferation of vascular smooth muscle cells (SMCs) causes restenosis in biomaterial vascular grafts. The purposes of this study were to establish a suspension culture system for SMCs by using a novel substrate, low-acyl gellan gum (GG) and to maintain SMCs in a state of growth inhibition. When SMCs were cultured in suspension with GG, their proliferation was inhibited. Their viability was 70% at day 2, which was maintained at more than 50% until day 5. In contrast, the viability of cells cultured in suspension without GG was 5.6% at day 2. By cell cycle analysis, the ratio of SMCs in the S phase when cultured in suspension with GG was lower than when cultured on plastic plates. In SMCs cultured in suspension with GG, the ratio of phosphorylated retinoblastoma (Rb) protein to Rb protein was decreased and p27(Kip1) expression was unchanged in comparison with SMCs cultured on plastic plates. In addition, SMCs could be induced to proliferate again by changing the culture condition from suspension with GG to plastic plates. These results suggest that our established culturing method for SMCs is useful to maintain SMCs in a state of growth inhibition with high viability.

  5. Organotypic Cultures of Intervertebral Disc Cells: Responses to Growth Factors and Signaling Pathways Involved

    Directory of Open Access Journals (Sweden)

    Harris Pratsinis


    Full Text Available Intervertebral disc (IVD degeneration is strongly associated with low back pain, a major cause of disability worldwide. An in-depth understanding of IVD cell physiology is required for the design of novel regenerative therapies. Accordingly, aim of this work was the study of IVD cell responses to mitogenic growth factors in a three-dimensional (3D organotypic milieu, comprising characteristic molecules of IVD’s extracellular matrix. In particular, annulus fibrosus (AF cells were cultured inside collagen type-I gels, while nucleus pulposus (NP cells in chondroitin sulfate A (CSA supplemented collagen gels, and the effects of Platelet-Derived Growth Factor (PDGF, basic Fibroblast Growth Factor (bFGF, and Insulin-Like Growth Factor-I (IGF-I were assessed. All three growth factors stimulated DNA synthesis in both AF and NP 3D cell cultures, with potencies similar to those observed previously in monolayers. CSA supplementation inhibited basal DNA synthesis rates, without affecting the response to growth factors. ERK and Akt were found to be phosphorylated following growth factor stimulation. Blockade of these two signaling pathways using pharmacologic inhibitors significantly, though not completely, inhibited growth factor-induced DNA synthesis. The proposed culture systems may prove useful for further in vitro studies aiming at future interventions for IVD regeneration.

  6. Batch and fed-batch production of butyric acid by Clostridium butyricum ZJUCB

    Institute of Scientific and Technical Information of China (English)

    HE Guo-qing; KONG Qing; CHEN Qi-he; RUAN Hui


    The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that cultivation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB.

  7. Effects of Basic Fibroblast Growth Factor and Insulin-like Growth Factor on Cultured Cartilage Cells from Skate Raja porasa

    Institute of Scientific and Technical Information of China (English)

    樊廷俊; 晋凌云; 汪小锋


    Effects of basic fibroblast growth factor (bFGF) and insulin-like growth factor II (IGF-II) on cartilage cells from proboscis of skate, Raja porasa Günther, were investigated in this study. The cartilage cells were cultured in 20% FBS-supplemented MEM medium at 24℃. Twelve hours after culture initiation, the cartilage cells were treated with bFGF and IGF-II at different concentration combinations. It was found that 20 ng/ml of bFGF or 80 ng/ml of IGF-II was enough to have obvious stimulating effect on the growth and division of skate cartilage cells. Test of bFGF and IGF-II together, revealed that 20 ng/ml of bFGF and 80 ng/ml of IGF-II together had the best stimulating effect on the growth and division of skate cartilage cells. The cartilage cells cultured could form a monolayer at day 7.

  8. Glucoamylase production in batch, chemostat and fed-batch cultivations by an industrial strain of Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Henrik; Beyer, Michael; Nielsen, Jens


    The Aspergillus niger strain BO-1 was grown in batch, continuous (chemostat) and fed-batch cultivations in order to study the production of the extracellular enzyme glucoamylase under different growth conditions. In the pH range 2.5-6.0, the specific glucoamylase productivity and the specific...

  9. Religious Orders and Growth through Cultural Change in Pre-Industrial England

    DEFF Research Database (Denmark)

    Andersen, Thomas Barnebeck; Bentzen, Jeanet; Dalgaard, Carl-Johan

    We advance the hypothesis that cultural values such as high work ethic and thrift, “the Protestant ethic” according to Max Weber, may have been diffused long before the Reformation, thereby importantly affecting the pre-industrial growth record. The source of pre-Reformation Protestant ethic, acc......-industrial development of England may thus have been propelled by a process of growth through cultural change.......We advance the hypothesis that cultural values such as high work ethic and thrift, “the Protestant ethic” according to Max Weber, may have been diffused long before the Reformation, thereby importantly affecting the pre-industrial growth record. The source of pre-Reformation Protestant ethic...

  10. Dynamic Batch Bayesian Optimization

    CERN Document Server

    Azimi, Javad; Fern, Xiaoli


    Bayesian optimization (BO) algorithms try to optimize an unknown function that is expensive to evaluate using minimum number of evaluations/experiments. Most of the proposed algorithms in BO are sequential, where only one experiment is selected at each iteration. This method can be time inefficient when each experiment takes a long time and more than one experiment can be ran concurrently. On the other hand, requesting a fix-sized batch of experiments at each iteration causes performance inefficiency in BO compared to the sequential policies. In this paper, we present an algorithm that asks a batch of experiments at each time step t where the batch size p_t is dynamically determined in each step. Our algorithm is based on the observation that the sequence of experiments selected by the sequential policy can sometimes be almost independent from each other. Our algorithm identifies such scenarios and request those experiments at the same time without degrading the performance. We evaluate our proposed method us...

  11. Growth and antrum formation of bovine primary follicles in long-term culture in vitro. (United States)

    Sun, Jing; Li, Xiangdong


    Successful antral formation in vitro from bovine preantral follicles (145-170 μm) has been described previously, but antrum formation from the primary follicle (50-70 μm) has not yet been achieved in vitro. The aim of the study was to establish an optimal culture system supporting the growth and maturation of bovine primary follicles (50-70 μm) in vitro. Bovine primary follicles were cultured in a three-dimensional culture system for 13 or 21 days in alpha-minimum essential medium. Various treatments including follicle stimulating hormone (FSH), luteinizing hormone (LH), 17β-estradiol (E2), basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were tested. The follicular diameter and antrum formation rate were recorded, and follicular maturation markers (P450 aromatase, CYP19A1; anti-Mullerian hormone, AMH; growth differentiation factor-9, GDF9; bone morphogenetic protein-15, BMP15; and type III transforming growth factor β receptor, TGFβR3) were analyzed by real-time RT-PCR. After 21 days of culture under each treatment condition, the follicular diameter was significantly enlarged in the presence of FSH + LH + E2 + bFGF or FSH + LH + E2 + bFGF + EGF (pculture, and the antral cavity formation rate was 16.7% and 33.3% by 21 days of culture, respectively. The expression of follicular maturation markers (CYP19A1, AMH, GDF9, BMP15 and TGFβR3) was significantly altered. We conclude that addition of 50 ng/ml bFGF +25 ng/ml EGF to media containing FSH + LH + E2 turned out to be the most effective optimized culture conditions to support the growth and maturation of bovine primary follicles in vitro.


    Directory of Open Access Journals (Sweden)

    Dagmara Kullačová


    Full Text Available Orobanchaceae family includes parasitic plants that attack many important food crops. Genus Phelipanche, belonging to this family is considered to cause high negative impact on food production. Developing Phelipanche plant must establish connection with the root of host plant, from which it receives all resources needed for further development. Nowadays big effort is directed to finding a reliable strategy to control parasitic plants. In vitro cultures of P. ramosa can be genetically manipulated and used for study of genes involved in host-parasite interactions. We established in vitro cultures of parasitic species Phelipanche ramosa on solid and liquid media in parallel. The obtained results point out that development of P. ramosa calli was origin specific. We tested the effect of antibiotic kanamycin on in vitro cultures of Phelipanche ramosa with aim to develop system for its genetic manipulation and selection of transgenic tissue using kanamycin- resistance approach. The selection pressure of kanamycin was stronger in liquid grown cultures. However, concentrations of kanamycin tested (up to 250 mg.l-1 did not ensured elimination of kanamycine non-resistant tissue. Tests of other candidate selection markers are currently in progress.

  13. Phenotypic and growth characterization of human mesenchymal stem cells cultured from permanent and deciduous teeth

    Directory of Open Access Journals (Sweden)

    Revathi Shekar


    Conclusions: Permanent and deciduous teeth are both viable sources of stem cells. The permanent teeth were easier to culture because of a lower chance of contamination with oral microflora. The growth characteristics of the cells obtained from both these sources were similar. However, there was a difference in the ratio of fibroblastoid cells to epithelioid cells between the cultures obtained from the permanent and deciduous teeth.

  14. Study on the Effects of Fed-batch Culture Method of Escherichia coli XD-12 Fermentation%流加培养方式对大肠杆菌XD-12发酵的影响研究

    Institute of Scientific and Technical Information of China (English)

    潘自皓; 顾薇; 潘扬


    [Objective] The research aimed to study the fed-batch culture method of Escherichia coli and enhance the fermentation concentration of transaminase donor.E. coli XD-12. [Method] The effects of carbon source feeding, nitrogen source feeding,feeding with pH control on E. coli fermentation were studied to obtain the optimal culture conditions. [ Result] The optimal cultural conditions lot transaminase-produeing E.coli were as Mows; temperature of 37 ℃ .agitation speed of 400 r/min,aeration quantity of 1.5 L/min,initial pH of medium of 7.0,for controlling fermentation process pH of 7.5,initial glucose concentration of 5 g/L,initial nitrogen source of 5 g/L peptone + 1.5 g/L beef extract, 120 g/L glucose solution was intermittently fed at an interval of 2 h after glucose concentration declined to 2 g/L And 15 g/L peptone and 4. 5 g/L beef extract were intermittently fed at an interval of 2 h after 8 hours. After culture 24 h under these conditions, the cell dry weight concentration of E.coli reached 9.66 g/L,104.7% higher than that in batch culture. [ Conclusion] This research was of an important realistic significance for reducing the production cost of preparing L-phenylalanine by enzyme method,enhancing the production efficiency of L-phenylala-nine,sufficing the increasing market requirements of L-phenylalanine.%[目的]研究大肠杆菌流加培养方式,提高转氨醇供体——大肠杆菌XD-12的发酵浓度.[方法]通过研究碳源流加、氮源流加、pH控制流加对发酵的影响,获得了优化的培养条件.[结果]产转氨酶大肠杆菌的最佳培养条件为:温度37℃,搅拌转速500 r/min,通气量1.5 L/min,培养基初始pH为7.0,控制发酵过程pH为7.5,初始葡萄糖浓度5 g/L,初始氮源为5g/L蛋白胨+1.5 g/L牛肉膏,从葡萄糖浓度下降为2 g/L开始每隔2h问歇流加120 g/L的糖,从8h起每隔2h间歇流加15 9/L蛋白胨+4.5 g/L牛肉膏.在此条件下培养24h,大肠杆菌的茵体干重浓度达9.66 g

  15. Religious Orders and Growth through Cultural Change in Pre-Industrial England

    DEFF Research Database (Denmark)

    Andersen, Thomas Barnebeck; Bentzen, Jeanet; Dalgaard, Carl-Johan;

    We advance the hypothesis that cultural values such as high work ethic and thrift, “the Protestant ethic” according to Max Weber, may have been diffused long before the Reformation, thereby importantly affecting the pre-industrial growth record. The source of pre-Reformation Protestant ethic......, according to the proposed theory, was the Catholic Order of Cistercians. Using county-level data for England we find empirically that the frequency of Cistercian monasteries influenced county-level comparative development until 1801; that is, long after the Dissolution of the Monasteries. The pre-industrial...... development of England may thus have been propelled by a process of growth through cultural change....

  16. Evaluation of growth yield of Spirulina (Arthrospira) sp. in photoautotrophic, heterotrophic and mixotrophic cultures. (United States)

    Chojnacka, Katarzyna; Zielińska, Agnieszka


    In microbial cultures, both cellular growth rate and yield (defined as the degree of substrate conversion into the biomass) are important. Although effect of culture conditions on growth kinetics has been well documented for various microbial strains, there is almost no literature concerning the effect of environmental conditions on growth equilibrium, expressed as biomass yield coefficients from substrate. The present paper discusses the effect of culture conditions: irradiance (physical substrate) and glucose concentration (chemical substrate) on biomass yield coefficients from two chemical substrates: glucose and nitrate-nitrogen in photoautotrophic, heterotrophic and mixotrophic culture of blue-green alga Spirulina (Arthrospira) sp. The efficiency of substrates incorporation into the biomass can be precisely determined only if the elemental composition of the biomass is known. The experimental results showed that culture conditions had a substantial influence on biomass yield coefficients (biomass yield from glucose and nitrate-nitrogen). It was found that, the increase of irradiance favoured increase of biomass yield coefficient from both, glucose and nitrate-nitrogen. However, in the case of yield from nitrogen in mixotrophic culture, the effect was opposite. The effect of glucose concentration was different: the higher the initial glucose concentration, the lower the biomass yield coefficients from chemical substrates.

  17. Impact of different culture media on hairy roots growth of Valeriana officinalis L.

    Directory of Open Access Journals (Sweden)



    Full Text Available Transformed hairy root cultures of Valeriana officinalis were established by infection with Agrobacterium rhizogenes strain ATCC 15834. To determine the effect of different media on the growth of V. officinalis hairy roots, MS, B5 media (1.0X and 0.5X strength, N6 medium and a modified MS medium without phytohormones were used. In addition, different NH4+ to NO3- ratios in MS medium were studied. The effects of these treatments were evaluated after 21 days of culture in relation to hairy root growth. B5 and ½ B5 media were the best basal media for hairy root growth. MS medium supplemented with a 20:20 ratio (mM of NH4+ to NO3- displayed highest growth rates and biomass yield in hairy root cultures. The present study demonstrated that the composition of culture medium and the ratio of different nitrogen sources have significant impact on the growth of V. officinalis hairy roots.

  18. Mycelial Biomass Exchange in Different Growth Stage Cultures of Flammulina velutipes

    Institute of Scientific and Technical Information of China (English)

    YU Changxia; CHEN Jianhua; CAO Hui; CHEN Mingjie; WANG Hong; WANG Yan


    Mycelial DNA was used as the parameter to determine fungal biomass in the cultivation substrate during the growth of Flammulina velutipes,strain FV.Substrate DNA levels,their effect on mycelial DNA extraction,and mycelial biomass in bottle cultures,were determined using a standard curve relating mycelial biomass with DNA content.Determination of mycelial biomass in samples collected from the upper,middle and lower layers of bottle cultures at different growth stages indicated a positive correlation between biomass and the ‘age’of the mycelium (i.e.growth period from inoculation and sample testing),and that biomass levels continually increased for a certain period after the substrate had been fully colonized.Mycelial biomass in the cultivation substrate decreased during fruit body growth,and occurred earlier and more rapidly in the upper layer compared with the middle and lower layers.

  19. Eucalyptus kraft black liquor enhances growth and productivity of Spirulina in outdoor cultures

    Energy Technology Data Exchange (ETDEWEB)

    Chauhan, V.S.; Singh, G.; Ramamurthy, V. [Thapar Institute of Engineering and Technology, Patiala (India)


    Mass cultivation of microalgae for commercial applications suffers from poor productivities when measured against laboratory results or theoretical projections. In an effort to reduce this gap it was discovered that addition of eucalyptus kraft black liquor (BL) enhanced biomass productivity in outdoor cultures of Spirulina by increasing growth rate by 38% and biomass yield by 43%. BL treatment resulted in elevation of nitrogen assimilating enzyme activities and efficiency of phosphate utilization. Analyses of forenoon and afternoon oxygen production rates (OPRs) indicated higher photosynthetic and respiratory activity in BL-treated cultures compared to untreated cultures. 20 refs., 1 fig., 2 tabs.

  20. Red Ginseng Extract Promotes the Hair Growth in Cultured Human Hair Follicles


    Park, Gyeong-Hun; Park, Ki-young; Cho, Hong-il; Lee, Sang-Min; Han, Ji Su; Won, Chong Hyun; Chang, Sung Eun; Lee, Mi Woo; Choi, Jee Ho; Moon, Kee Chan; Shin, Hyoseung; KANG, YONG JUNG; Lee, Dong Hun


    Ginseng has been shown to promote hair growth in several recent studies. However, its effects on human hair follicles and its mechanisms of action have not been sufficiently elucidated. This study aimed to investigate the hair growth-promoting effects of red ginseng extract (RGE) and its ginsenosides. The proliferative activities of cultured human hair follicles treated with RGE and ginsenoside-Rb1 were assessed using Ki-67 immunostaining. Their effects on isolated human dermal papilla cells ...

  1. Viability of mycelial growth of Coprinus Comatus in culture medium based on organic residues


    de Almeida, Lais Benes; Sales-Campos,Ceci; Melo de Carvalho, Cristiane Suely; de Almeida Minhoni, Marli Teixeira; Nogueira de Andrade, Meire Cristina


    The objective of this work was to evaluate the mycelial growth of the Coprinus comatus strain CCO 01/01 in culture based on organic residues of Saccharum officinarum (sugarcane bagasse), Citrus sinensis (orange bagasse), Ananas comosus (pineapple residues) and Musa sp. (banana leaf), supplemented with wheat bran in the proportions of 0, 10 and 20%, kept at 27 degrees C. The mycelial growth of C. comatus was evaluated daily by measurement of the diameter of the colony during seven days of incu...

  2. Automated screening of blood cultures with the Malthus microbiological growth analyser. (United States)

    Brown, D F; Warner, M; Taylor, C E; Warren, R E


    A total of 3347 blood cultures from patients in all hospital wards were examined on a Malthus microbiological growth analyser and by a conventional system. There was no significant difference in the total numbers of positive cultures of clinical importance between the two systems (p greater than 0.05). Staphylococcus aureus, however, was isolated more often by the conventional method (p less than 0.05). Failure of the automatic detection routine limited the potential of the Malthus system for earlier detection of positive cultures. Daily visual examination of Malthus curves and subculture of bottles not promptly attached to the apparatus were necessary to avoid missing some positive cultures. False positive rates were 13% for the Malthus system and 2% for the conventional system. The contamination rate was considerably lower in the Malthus system (p less than 0.001). Further development would be necessary for the apparatus to be acceptable for routine screening of blood cultures.


    Institute of Scientific and Technical Information of China (English)

    张蓓; 张克旭; 陈宁; 徐咏全


    It is well recognized that metabolic fluxes are the key variables that must be determined in orderto understand metabolic regulation and patterns. However, owing to difficulties in measuring the flux values,evaluation of metabolic fluxes has not been an integral part of the most metabolic studies. Flux values formetabolites of glycolysis (EMP), tricarboxylic acid (TCA) cycle, and hexose monophosphate (HMP) pathwaywas obtained for batch cultures of inosine overproducing Bacilus subtilis by combining the information from thestoichiometry of key biosynthetic reactions with the experimental data on uptake rate of glucose and formationrates of metabolic product and byproducts.%为了更好地掌握代谢规律和代谢方式,代谢流作为一个关键而重要的变量需要测定.然而,由于代谢流测定的困难,在许多代谢研究中代谢流并没有被完全应用.在肌苷高产菌枯草芽孢杆菌中,利用得到的实验数据包括葡萄糖的消耗速率和代谢主产物以及副产物的形成速率,利用代谢通量平衡模型,得到糖酵解、三羟酸循环及磷酸戊糖途径的代谢流,并对其进行了分析.

  4. Antipathogenic activity of probiotics against Salmonella Typhimurium and Clostridium difficile in anaerobic batch culture systems: is it due to synergies in probiotic mixtures or the specificity of single strains? (United States)

    Tejero-Sariñena, Sandra; Barlow, Janine; Costabile, Adele; Gibson, Glenn R; Rowland, Ian


    Probiotics are currently being investigated for prevention of infections caused by enteric pathogens. The aim of this in vitro study was to evaluate the influence of three single probiotics: Lactobacillus casei NCIMB 30185 (PXN 37), Lactobacillus acidophilus NCIMB 30184 (PXN 35), Bifidobacterium breve NCIMB 30180 (PXN 25) and a probiotic mixture containing the above strains plus twelve other strains belonging to the Lactobacillus, Bifidobacterium, Lactococcus, Streptococcus and Bacillus genera on the survival of Salmonella Typhimurium and Clostridium difficile using pH-controlled anaerobic batch cultures containing mixed faecal bacteria. Changes in relevant bacterial groups and effects of probiotic addition on survival of the two pathogens were assessed over 24 h. Quantitative analysis of bacterial populations revealed that there was a significant increase in lactobacilli and/or bifidobacteria numbers, depending on probiotic addition, compared with the control (no added probiotic). There was also a significant reduction in S. Typhimurium and C. difficile numbers in the presence of certain probiotics compared with controls. Of the probiotic treatments, two single strains namely L. casei NCIMB 30185 (PXN 37), and B. breve NCIMB 30180 (PXN 25) were the most potent in reducing the numbers of S. Typhimurium and C. difficile. In addition, the supplementation with probiotics into the systems influenced some fermentations parameters. Acetate was found in the largest concentrations in all vessels and lactate and formate were generally detected in higher amounts in vessels with probiotic addition compared to controls.

  5. Efficient estimation of the maximum metabolic productivity of batch systems

    Energy Technology Data Exchange (ETDEWEB)

    St. John, Peter C.; Crowley, Michael F.; Bomble, Yannick J.


    Production of chemicals from engineered organisms in a batch culture involves an inherent trade-off between productivity, yield, and titer. Existing strategies for strain design typically focus on designing mutations that achieve the highest yield possible while maintaining growth viability. While these methods are computationally tractable, an optimum productivity could be achieved by a dynamic strategy in which the intracellular division of resources is permitted to change with time. New methods for the design and implementation of dynamic microbial processes, both computational and experimental, have therefore been explored to maximize productivity. However, solving for the optimal metabolic behavior under the assumption that all fluxes in the cell are free to vary is a challenging numerical task. Previous studies have therefore typically focused on simpler strategies that are more feasible to implement in practice, such as the time-dependent control of a single flux or control variable.

  6. Cholera toxin expression by El Tor Vibrio cholerae in shallow culture growth conditions. (United States)

    Cobaxin, Mayra; Martínez, Haydee; Ayala, Guadalupe; Holmgren, Jan; Sjöling, Asa; Sánchez, Joaquín


    Vibrio cholerae O1 classical, El Tor and O139 are the primary biotypes that cause epidemic cholera, and they also express cholera toxin (CT). Although classical V. cholerae produces CT in various settings, the El Tor and O139 strains require specific growth conditions for CT induction, such as the so-called AKI conditions, which consist of growth in static conditions followed by growth under aerobic shaking conditions. However, our group has demonstrated that CT production may also take place in shallow static cultures. How these type of cultures induce CT production has been unclear, but we now report that in shallow culture growth conditions, there is virtual depletion of dissolved oxygen after 2.5 h of growth. Concurrently, during the first three to 4 h, endogenous CO2 accumulates in the media and the pH decreases. These findings may explain CT expression at the molecular level because CT production relies on a regulatory cascade, in which the key regulator AphB may be activated by anaerobiosis and by low pH. AphB activation stimulates TcpP synthesis, which induces ToxT production, and ToxT directly stimulates ctxAB expression, which encodes CT. Importantly, ToxT activity is enhanced by bicarbonate. Therefore, we suggest that in shallow cultures, AphB is activated by initial decreases in oxygen and pH, and subsequently, ToxT is activated by intracellular bicarbonate that has been generated from endogenous CO2. This working model would explain CT production in shallow cultures and, possibly, also in other growth conditions.

  7. Biological hydrogen production measured in batch anaerobic respirometers. (United States)

    Logan, Bruce E; Oh, Sang-Eun; Kim, In S; Van Ginkel, Steven


    The biological production of hydrogen from the fermentation of different substrates was examined in batch tests using heat-shocked mixed cultures with two techniques: an intermittent pressure release method (Owen method) and a continuous gas release method using a bubble measurement device (respirometric method). Under otherwise identical conditions, the respirometric method resulted in the production of 43% more hydrogen gas from glucose than the Owen method. The lower conversion of glucose to hydrogen using the Owen protocol may have been produced by repression of hydrogenase activity from high partial pressures in the gastight bottles, but this could not be proven using a thermodynamic/rate inhibition analysis. In the respirometric method, total pressure in the headspace never exceeded ambient pressure, and hydrogen typically composed as much as 62% of the headspace gas. High conversion efficiencies were consistently obtained with heat-shocked soils taken at different times and those stored for up to a month. Hydrogen gas composition was consistently in the range of 60-64% for glucose-grown cultures during logarithmic growth but declined in stationary cultures. Overall, hydrogen conversion efficiencies for glucose cultures were 23% based on the assumption of a maximum of 4 mol of hydrogen/ mol of glucose. Hydrogen conversion efficiencies were similar for sucrose (23%) and somewhat lower for molasses (15%) but were much lower for lactate (0.50%) and cellulose (0.075%).

  8. Reactive oxygen species are involved in BMP-induced dendritic growth in cultured rat sympathetic neurons. (United States)

    Chandrasekaran, Vidya; Lea, Charlotte; Sosa, Jose Carlo; Higgins, Dennis; Lein, Pamela J


    Previous studies have shown that bone morphogenetic proteins (BMPs) promote dendritic growth in sympathetic neurons; however, the downstream signaling molecules that mediate the dendrite promoting activity of BMPs are not well characterized. Here we test the hypothesis that reactive oxygen species (ROS)-mediated signaling links BMP receptor activation to dendritic growth. In cultured rat sympathetic neurons, exposure to any of the three mechanistically distinct antioxidants, diphenylene iodinium (DPI), nordihydroguaiaretic acid (NGA) or desferroxamine (DFO), blocked de novo BMP-induced dendritic growth. Addition of DPI to cultures previously induced with BMP to extend dendrites caused dendritic retraction while DFO and NGA prevented further growth of dendrites. The inhibition of the dendrite promoting activity of BMPs by antioxidants was concentration-dependent and occurred without altering axonal growth or neuronal cell survival. Antioxidant treatment did not block BMP activation of SMAD 1,5 as determined by nuclear localization of these SMADs. While BMP treatment did not cause a detectable increase in intracellular ROS in cultured sympathetic neurons as assessed using fluorescent indicator dyes, BMP treatment increased the oxygen consumption rate in cultured sympathetic neurons as determined using the Seahorse XF24 Analyzer, suggesting increased mitochondrial activity. In addition, BMPs upregulated expression of NADPH oxidase 2 (NOX2) and either pharmacological inhibition or siRNA knockdown of NOX2 significantly decreased BMP-7 induced dendritic growth. Collectively, these data support the hypothesis that ROS are involved in the downstream signaling events that mediate BMP7-induced dendritic growth in sympathetic neurons, and suggest that ROS-mediated signaling positively modulates dendritic complexity in peripheral neurons.

  9. An automated blood culture system: the detection of anaerobic bacteria using a Malthus Microbiological Growth Analyser. (United States)

    McMaster, J P; Barr, J G; Campbell, R R; Bennett, R B; Smyth, E T


    The Malthus Microbiological Growth Analyser has proved to be sensitive in detecting conductivity changes due to anaerobic metabolism in a number of widely used blood culture media. Freshly prepared cooked meat media and Thiol medium yielded the greatest gross conductivity changes, and were more sensitive of anaerobic metabolism than other media. Failure of the instrument to detect anaerobic metabolism was a problem particularly associated with growth in the thioglycollate medium. False positive detections of growth were attributed to a number of factors including electrode instability (6.0%) and bacterial contamination (8.75%).

  10. Growth kinetics and physiological behavior of co-cultures of Saccharomyces cerevisiae and Kluyveromyces lactis, fermenting carob sugars extracted with whey. (United States)

    Rodrigues, B; Lima-Costa, M E; Constantino, A; Raposo, S; Felizardo, C; Gonçalves, D; Fernandes, T; Dionísio, L; Peinado, J M


    Alcoholic fermentation of carob waste sugars (sucrose, glucose and fructose) extracted with cheese whey, by co-cultures of Saccharomyces cerevisiae and Kluyveromyces lactis has been analyzed. Growth and fermentation of S. cerevisiae in the carob-whey medium showed an inhibition of about 30% in comparison with water-extracted carob. The inhibition of K. lactis on carob-whey was greater (70%) when compared with the whey medium alone, due to osmolarity problems. Oxygen availability was a very important factor for K. lactis, influencing its fermentation performance. When K. lactis was grown alone on carob-whey medium, lactose was always consumed first, and glucose and fructose were consumed afterwards, only at high aeration conditions. In co-culture with S. cerevisiae, K. lactis was completely inhibited and, at low aeration, died after 3 days; at high aeration this culture could survive but growth and lactose fermentation were only recovered after S. cerevisiae became stationary. To overcome the osmolarity and K. lactis' oxygen problems, the medium had to be diluted and a sequential fermentative process was designed in a STR-3l reactor. K. lactis was inoculated first and, with low aeration (0.13vvm), consumed all the lactose in 48h. Then S. cerevisiae was inoculated, consuming the total of the carob sugars, and producing ethanol in a fed-batch regime. The established co-culture with K. lactis increased S. cerevisiae ethanol tolerance. This fermentation process produced ethanol with good efficiency (80g/l final concentration and a conversion factor of 0.4g ethanol/g sugar), eliminating all the sugars of the mixed waste. These efficient fermentative results pointed to a new joint treatment of agro-industrial wastes which may be implemented successfully, with economic and environmental sustainability for a bioethanol industrial proposal.

  11. Growth of Crassostrea gasar cultured in marine and estuary environments in Brazilian waters

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    Gustavo Ruschel Lopes


    Full Text Available The objective of this work was to evaluate the growth of the mangrove oyster Crassostrea gasar cultured in marine and estuarine environments. Oysters were cultured for 11 months in a longline system in two study sites - São Francisco do Sul and Florianópolis -, in the state of Santa Catarina, Southern Brazil. Water chlorophyll-α concentration, temperature, and salinity were measured weekly. The oysters were measured monthly (shell size and weight gain to assess growth. At the end of the culture period, the average wet flesh weight, dry flesh weight, and shell weight were determined, as well as the distribution of oysters per size class. Six nonlinear models (logistic, exponential, Gompertz, Brody, Richards, and Von Bertalanffy were adjusted to the oyster growth data set. Final mean shell sizes were higher in São Francisco do Sul than in Florianópolis. In addition, oysters cultured in São Francisco do Sul were more uniformly distributed in the four size classes than those cultured in Florianópolis. The highest average values of wet flesh weight and shell weight were observed in São Francisco do Sul, whereas dry flesh weight did not differ between the sites. The estuary environment is more promising for the cultivation of oysters.

  12. Serum obtained from rats after partial hepatectomy enhances growth of cultured colon carcinoma cells

    NARCIS (Netherlands)

    de Jong, KP; Brouwers, MAM; van Veen, ML; Brinker, M; de Vries, EGE; Daemen, T; Scherphof, GL; Slooff, MJH


    Tumour-bearing rats were randomized to a 70% partial hepatectomy or a sham operation. At days 1, 3 or 14, portal and systemic serum was obtained and colon carcinoma cells were cultured in the presence of 5, 10, 20 or 50% serum. Proliferation and epidermal growth factor receptor (EGFr) expression was

  13. Growth of plant root cultures in liquid- and gas-dispersed reactor environments. (United States)

    McKelvey, S A; Gehrig, J A; Hollar, K A; Curtis, W R


    The growth of Agrobacterium transformed "hairy root" cultures of Hyoscyamus muticus was examined in various liquid- and gas-dispersed bioreactor configurations. Reactor runs were replicated to provide statistical comparisons of nutrient availability on culture performance. Accumulated tissue mass in submerged air-sparged reactors was 31% of gyratory shake-flask controls. Experiments demonstrate that poor performance of sparged reactors is not due to bubble shear damage, carbon dioxide stripping, settling, or flotation of roots. Impaired oxygen transfer due to channeling and stagnation of the liquid phase are the apparent causes of poor growth. Roots grown on a medium-perfused inclined plane grew at 48% of gyratory controls. This demonstrates the ability of cultures to partially compensate for poor liquid distribution through vascular transport of nutrients. A reactor configuration in which the medium is sprayed over the roots and permitted to drain down through the root tissue was able to provide growth rates which are statistically indistinguishable (95% T-test) from gyratory shake-flask controls. In this type of spray/trickle-bed configuration, it is shown that distribution of the roots becomes a key factor in controlling the rate of growth. Implications of these results regarding design and scale-up of bioreactors to produce fine chemicals from root cultures are discussed.

  14. The role of DNA amplification and cultural growth in complicated acute appendicitis

    Directory of Open Access Journals (Sweden)

    Francesca Tocchioni


    Full Text Available Bacterial growth of peritoneal fluid specimens obtained during surgical procedures for acute appendicitis may be useful to optimize further antibiotic therapy in complicated cases. DNA amplification represents a fast technique to detect microbial sequences. We aimed to compare the potential of DNA amplification versus traditional bacterial growth culture highlighting advantages and drawbacks in a surgical setting. Peritoneal fluid specimens were collected during surgery from 36 children who underwent appendectomy between May and December 2012. Real-time polymerase chain reaction (RT-PCR and cultures were performed on each sample. RT-PCR showed an amplification of 16S in 18/36 samples, Escherichia coli (in 7 cases, Pseudomonas aeruginosa (3, Fusobacterium necrophorum (3, Adenovirus (2, E.coli (1, Klebsiella pneumoniae (1, Serratia marcescens/Enterobacter cloacae (1. Bacterial growth was instead observed only in four patients (3 E.coli and 1 P.aeruginosa and Bacteroides ovatus. Preoperative C-reactive protein and inflammation degree, the most reliable indicators of bacterial translocation, were elevated as expected. DNA amplification was a quick and useful method to detect pathogens and it was even more valuable in detecting aggressive pathogens such as anaerobes, difficult to preserve in biological cultures; its drawbacks were the lack of biological growths and of antibiograms. In our pilot study RT-PCR and cultures did not influence the way patients were treated.

  15. Photophysiological variability of microphytobenthic diatoms after growth in different types of culture conditions

    NARCIS (Netherlands)

    Forster, R.M.; Martin-Jézéquel, V.R.


    Microphytobenthic diatoms have great ecological importance in estuarine and coastal marine ecosystenis, yet many aspects of their physiology have not been investigated under controlled conditions. This work describes patterns in growth rates and photosynthesis in different types of culture for sever

  16. Teaching Population Growth Using Cultures of Vinegar Eels, "Turbatrix aceti" (Nematoda) (United States)

    Wallace, Robert L.


    A simple laboratory exercise is presented that follows the population growth of the common vinegar eel, "Turbatrix aceti" (Nematoda), in a microcosm using a simple culture medium. It lends itself to an exercise in a single semester course. (Contains 4 figures.)

  17. The Role of DNA Amplification and Cultural Growth in Complicated Acute Appendicitis (United States)

    Tocchioni, Francesca; Tani, Chiara; Bartolini, Laura; Moriondo, Maria; Nieddu, Francesco; Pecile, Patrizia; Azzari, Chiara; Messineo, Antonio; Ghionzoli, Marco


    Bacterial growth of peritoneal fluid specimens obtained during surgical procedures for acute appendicitis may be useful to optimize further antibiotic therapy in complicated cases. DNA amplification represents a fast technique to detect microbial sequences. We aimed to compare the potential of DNA amplification versus traditional bacterial growth culture highlighting advantages and drawbacks in a surgical setting. Peritoneal fluid specimens were collected during surgery from 36 children who underwent appendectomy between May and December 2012. Real-time polymerase chain reaction (RT-PCR) and cultures were performed on each sample. RT-PCR showed an amplification of 16S in 18/36 samples, Escherichia coli (in 7 cases), Pseudomonas aeruginosa (3), Fusobacterium necrophorum (3), Adenovirus (2), E.coli (1), Klebsiella pneumoniae (1), Serratia marcescens/Enterobacter cloacae (1). Bacterial growth was instead observed only in four patients (3 E.coli and 1 P.aeruginosa and Bacteroides ovatus). Preoperative C-reactive protein and inflammation degree, the most reliable indicators of bacterial translocation, were elevated as expected. DNA amplification was a quick and useful method to detect pathogens and it was even more valuable in detecting aggressive pathogens such as anaerobes, difficult to preserve in biological cultures; its drawbacks were the lack of biological growths and of antibiograms. In our pilot study RT-PCR and cultures did not influence the way patients were treated. PMID:27777701

  18. Relationship among growth parameters for Clostridium butyricum, hydA gene expression, and biohydrogen production in a sucrose-supplemented batch reactor. (United States)

    Wang, Mei-Yun; Olson, Betty H; Chang, Jo-Shu


    This study was undertaken to identify the relationship between the performance of dark H2 fermentation and expression of the key functional gene (i.e., hydrogenase gene) involved in the bioH2 production process. Clostridium butyricum CGS5 isolated from anaerobic sewage sludge was used as the model strain for this study. Copy number of the hydrogenase gene (hydA) and mRNA transcripts (cDNA hydA) (after amplification) and the total DNA and RNA (before amplification) were measured over the course of the growth of strain CGS5. Cell concentration was also determined by optical density and converted to dry weight. After amplification, the hydA gene increased 1,500-fold during late exponential growth phase after normalization to the copy number at time 0, and cDNA from mRNA transcripts of hydA also increased 500-fold after normalization. mRNA transcripts of hydA lagged behind the increase of total DNA and RNA, and increases in hydA more closely mimicked those of total DNA. Increases in both of these parameters corresponded with hydrogen production. Transcripts of 16s ribosomal RNA reached a maximum value earlier (38 h) than did those of hydA (47 h). All molecular characteristics matched those for sucrose utilization, growth, and hydrogen production. These experiments indicated that transcription as measured by cDNA can be related to hydrogen production and possesses the potential to be used as tool for process control.

  19. Inhibition of growth of Toxoplasma gondii in cultured fibroblasts by human recombinant gamma interferon. (United States)

    Pfefferkorn, E R; Guyre, P M


    The growth of Toxoplasma gondii in cultured human fibroblasts was inhibited by recombinant human gamma interferon at concentrations of 8 to 16 U/ml. The interferon was titrated by observing a total inhibition of parasite plaque formation 7 days after infection. Inhibition of the growth of T. gondii in the early days after infection was measured by marked reductions in the incorporation of radioactive uracil, a precursor that can only be used by the parasites. This assay showed that when cells were pretreated with gamma interferon for 1 day and then infected, inhibition of T. gondii growth could be readily detected 1 or 2 days after infection. When the pretreatment was omitted and parasites and gamma interferon were added at the same time, no inhibition of parasite growth could be detected 1 day later, although it was apparent after 2 days. Cultures from which the gamma interferon had been removed by washing after a 1-day treatment showed inhibition of T. gondii growth. Gamma interferon had no effect on the viability of extracellular parasites, but it did inhibit the synthesis of host cell RNA and protein by ca. 50% 3 days after treatment. This degree of inhibition is unlikely, of itself, to compromise the growth of T. gondii. Recombinant alpha and beta interferons had no effect on the growth of T. gondii. Images PMID:6425215

  20. Effects of transgenic soybean on growth and phosphorus acquisition in mixed culture system

    Institute of Scientific and Technical Information of China (English)

    Jianna Xie; Jia Zhou; Xiurong Wang; Hong Liao


    Transgenic soybean plants overexpressing the Arabidopsis purple acid phosphatase gene AtPAP15 (OXp) or the soybean expansin gene GmEXPB2 (OXe) can improve phosphorous (P) efficiency in pure culture by increasing Apase secretion or changing root morphology. In this study, soybean‐soybean mixed cultures were employed to il uminate P acquisition among plants in mixed stands of transgenic and wild‐type soybean. Our results showed that transgenic soybean plants were much more competitive, and had greater growth and P uptake than wild‐type soybean in mixed culture in both low P calcareous and acid soils. Furthermore, OXe plants had an advantage in calcareous soils when mixed with OXp, whereas the latter performed much better in acid soils. In soybean‐maize mixed culture, transgenic soybean had no impact on maize growth compared to controls in both acid and calcareous soils with different P conditions. As for soybean in mixed culture, OXp plants had no significant advantages regardless of P availability or soil type, while P efficiency improved in OXe in calcareous soils compared to controls. These results imply that physiological traits could be easily affected by the mixed maize. Transgenic soybean plants with enhanced root traits had more competitive advantages than those with improved root physiology in mixed culture.

  1. Enhanced algae growth in both phototrophic and mixotrophic culture under blue light. (United States)

    Das, Probir; Lei, Wang; Aziz, Siti Sarah; Obbard, Jeffrey Philip


    Biomass productivity and fatty acid methyl esters (FAME) derived from intracellular lipid of a Nannochloropsis sp. isolated from Singapore's coastal waters were studied under different light wavelengths and intensities. Nannochloropsis sp., was grown in both phototrophic and mixotrophic (glycerol as the carbon source) culture conditions in three primary monochromatic light wavelengths, i.e., red, green and blue LEDs, and also in white LED. The maximum specific growth rate (μ) for LEDs was blue>white>green>red. Nannochloropsis sp. achieved a μ of 0.64 and 0.66 d(-1) in phototrophic and mixotrophic cultures under blue lighting, respectively. The intracellular fatty acid composition of Nannochloropsis sp. varied between cultures exposed to different wavelengths, although the absolute fatty acid content did differ significantly. Maximum FAME yield from Nannochloropsis sp. was 20.45% and 15.11% of dry biomass weight equivalent under photo- and mixotrophic culture conditions respectively for cultures exposed to green LED (550 nm). However, maximum volumetric FAME yield was achieved for phototrophic and mixotrophic cultures (i.e., 55.13 and 111.96 mg/l, respectively) upon cell exposure to blue LED (470 nm) due to highest biomass productivity. It was calculated that incremental exposure of light intensity over the cell growth cycle saves almost 20% of the energy input relative to continuous illumination for a given light intensity.

  2. Effects of outdoor cultures on the growth and lipid production of Phaeodactylum tricornutum using closed photobioreactors. (United States)

    Santos-Ballardo, David U; Rendón-Unceta, María Del Carmen; Rossi, Sergio; Vázquez-Gómez, Rosa; Hernández-Verdugo, Sergio; Valdez-Ortiz, Angel


    One of the principal challenges for large scale production of microalgae is the high costs of biomass production. Aiming for minimize this problem, microalgal biodiesel production should focus on outdoors cultures, using available solar light and allowing lower energy cost process. Testing species that proved to be common and easy to culture may be a good approach in this process. The present work reports indoor-outdoor cultures of Phaeodactylum tricornutum using different bioreactors types, using cell growth, biochemical composition, and the profiles of the fatty acids produced as the parameters to test the optimization processes. The results show that the use of outdoor cultures is a good choice to obtain P. tricornutum biomass with a good potential for biodiesel production. The microalgae produced reached better growth efficiency, major lipid content and showed an increment in the percentage of saturated fatty acids (required on the biodiesel production) respect indoor cultures. These results are important to show the relevance of using outdoor cultures as a way to improve the efficiency and the energetic balance of the biodiesel production with P. tricornutum algae.

  3. Growth kinetics of Isochrysis galbana in cultures containing sublethal concentrations of mercuric chloride

    Energy Technology Data Exchange (ETDEWEB)

    Davies, A.G.


    Measurements of the effect of mercuric chloride upon the growth of Isochrysis galbana in culture are reported. The mercury was concentrated by the plant cells to a very high level, even sublethal intracellular concentrations being of the order of 1 g/l. It is shown that the specific growth rate of the cells in the presence of mercury was reduced linearly by increasing intracellular levels of the metal, thus providing a convenient relationship between growth and mercury uptake. The intracellular concentration which just prevented growth (about 2 g/l) was not instantly lethal, and the loss of mercury from the cultures due to volatilizaton caused by chemical reduction of the ionic form eventually allowed growth to start again giving final population levels approaching that in the control. The presence of mercury in the cells was found to have a considerable effect upon the mean cell volumes in the cultures causing almost a doubling of size at the highest sublethal concentrations examined; this was probably due to combination of the metal with the sulphur-containing compounds which are known to be of importance in the process of cell division.

  4. Characterization of growth hormone and prolactin produced by human pituitary in culture. (United States)

    Skyler, J S; Rogol, A D; Lovenberg, W; Knazek, R A


    Fragments of a pituitary tumor from a patient with acromegaly were grown in tissue culture. The tumor secreted both growth hormone and prolactin,which were recovered in high concentrations. The nonpurified hormones were characterized and compared to their respective counterparts obtained by extraction from normal pituitaries obtained at autopsy. The tissue culture and pituitary extracted hormones were eluted from Sephadex G-100 with the same partition coefficients. Growth hormone from both sources showed parallel dose-response displacement curves, by logit-log transformation, in both specific immunoassay and in a specific lymphocyte binding assay. Prolactin from both sources was compared in specific immunoassay using three different antisera. Parallel logit-log displacement curves were seen with one antiserum, while the other two antisera yielded non-parallel curves, indicating structural differences between prolactin from the two sources. Quantitative polyacrylamide gel electrophoresis was performed using multiphasic buffer systems previously developed for characterization of each hormone. By the criteria of joint 95% confidence envelopes of retardation co-efficient and relative free mobility, tissue culture growth hormone and prolactin were indistinguishable from their pituitary-extracted counterparts. This study demonstrates that, prior to purification, tissue culture derived hormone can be characterized by multiple criteria and compared to a standard preparation. Structural differences can be detected, as in the case of prolactin. When the hormones are indistinguishable, as in the case of growth hormone, it becomes worthwhile to increase the scale of tissue cultured production, with the prospect that tissue culture may serve as a source of hormone for both experimental and therapeutic use.

  5. Fed-batch process for the psychrotolerant marine bacterium Pseudoalteromonas haloplanktis

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    Lalk Michael


    Full Text Available Abstract Background Pseudoalteromonas haloplanktis is a cold-adapted γ-proteobacterium isolated from Antarctic sea ice. It is characterized by remarkably high growth rates at low temperatures. P. haloplanktis is one of the model organisms of cold-adapted bacteria and has been suggested as an alternative host for the soluble overproduction of heterologous proteins which tend to form inclusion bodies in established expression hosts. Despite the progress in establishing P. haloplanktis as an alternative expression host the cell densities obtained with this organism, which is unable to use glucose as a carbon source, are still low. Here we present the first fed-batch cultivation strategy for this auspicious alternative expression host. Results The key for the fed-batch cultivation of P. haloplanktis was the replacement of peptone by casamino acids, which have a much higher solubility and allow a better growth control. In contrast to the peptone medium, on which P. haloplanktis showed different growth phases, on a casamino acids-containing, phosphate-buffered medium P. haloplanktis grew exponentially with a constant growth rate until the stationary phase. A fed-batch process was established by feeding of casamino acids with a constant rate resulting in a cell dry weight of about 11 g l-1 (OD540 = 28 which is a twofold increase of the highest densities which have been obtained with P. haloplanktis so far and an eightfold increase of the density obtained in standard shake flask cultures. The cell density was limited in the fed-batch cultivation by the relatively low solubility of casamino acids (about 100 g l-1, which was proven by pulse addition of casamino acid powder which increased the cell density to about 20 g l-1 (OD540 = 55. Conclusion The growth of P. haloplanktis to higher cell densities on complex medium is possible. A first fed-batch fermentation strategy could be established which is feasible to be used in lab-scale or for industrial

  6. Influence of Environmental Factors and Medium Composition on Vibrio gazogenes Growth and Prodigiosin Production (United States)

    Allen, Garry R.; Reichelt, John L.; Gray, Peter P.


    Vibrio gazogenes ATCC 29988 growth and prodigiosin synthesis were studied in batch culture on complex and defined media and in chemostat cultures on defined medium. In batch culture on complex medium, a maximum growth rate of 0.75 h−1 and a maximum prodigiosin concentration of 80 ng of prodigiosin · mg of cell protein−1 were observed. In batch culture on defined medium, maximum growth rates were lower (maximum growth rate, 0.40 h−1), and maximum prodigiosin concentrations were higher (1,500 ng · mg of protein−1). In batch culture on either complex or defined medium, growth was characterized by a period of logarithmic growth followed by a period of linear growth; on either medium, prodigiosin biosynthesis was maximum during linear growth. In batch culture on defined medium, the initial concentration of glucose optimal for growth and pigment production was 3.0%; higher levels of glucose suppressed synthesis of the pigment. V. gazogenes had an absolute requirement for Na+; optimal growth occurred in the presence of 100 mM NaCl. Increases in the concentration of Na+ up to 600 mM resulted in further increases in the concentration of pigment in the broth. Prodigiosin was synthesized at a maximum level in the presence of inorganic phosphate concentrations suboptimal for growth. Concentrations of KH2PO4 above 0.4 mM caused decreased pigment synthesis, whereas maximum cell growth occurred at 1.0 mM. Optimal growth and pigment production occurred in the presence of 8 to 16 mg of ferric ion · liter−1, with higher concentrations proving inhibitory to both growth and pigment production. Both growth and pigment production were found to decrease with increased concentrations of p-aminobenzoic acid. The highest specific concentration of prodigiosin (3,480 ng · mg protein−1) was observed in chemostat cultures at a dilution rate of 0.057 h−1. The specific rate of prodigiosin production at this dilution rate was approximately 80% greater than that observed in batch

  7. Unusal pattern of product inhibition: batch acetic acid fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Bar, R.; Gainer, J.L.; Kirwan, D.J.


    The limited tolerance of microorganisms to their metabolic products results in inhibited growth and product formation. The relationship between the specific growth rate, micro, and the concentration of an inhibitory product has been described by a number of mathematical models. In most cases, micro was found to be inversely proportional to the product concentration and invariably the rate of substrate utilization followed the same pattern. In this communication, the authors report a rather unusual case in which the formation rate of a product, acetic acid, increased with a decreasing growth rate of the microorganism, Acetobacter aceti. Apparently, a similar behavior was mentioned in a review report with respect to Clostridium thermocellum in a batch culture but was not published in the freely circulating literature. The fermentation of ethanol to acetic acid, C/sub 2/H/sub 5/OH + O/sub 2/ = CH/sub 3/COOH + H/sub 2/O is clearly one of the oldest known fermentations. Because of its association with the commercial production of vinegar it has been a subject of extensive but rather technically oriented studies. Suprisingly, the uncommon uncoupling between the inhibited microbial growth and the product formation appears to have been unnoticed. 13 references.

  8. Biochemical and morphological changes during the growth kinetics of Araucaria angustifolia suspension cultures

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    André Luis Wendt dos Santos


    Full Text Available Embryogenic cultures of Araucaria angustifolia were established in a BM liquid medium supplemented with 2 µM 2,4-dichlorophenoxyacetic acid, 1 µM 6-benzylaminopurine and 1 µM kinetin (BM2 and in a BM medium free of growth regulators (BM0. During 42 days in culture, the cell growth pattern of both cultures was similar. The pH of the culture medium of both BM0 and BM2 underwent progressive reduction during culture time. For both the embryogenic cultures a preferential uptake of glucose in the late stages of cell growth kinetics was observed. The extracellular protein content was similar for both the embryogenic cultures. Acetocarmine and Evan's blue double stain showed major differences for early somatic embryo organisation, in which only the embryogenic culture grown in a liquid culture medium free of plant growth regulators showed the presence of bipolar somatic pro-embryos.Culturas embriogênicas de Araucaria angustifolia foram estabelecidas em meio de cultura líquido BM suplementado com 2 µM Ácido 2,4 Diclorofenoxiacético, 1 µM 6-Benzilaminopurina e 1 µM Cinetina (BM2 e em meio BM isento de reguladores de crescimento (BM0. Durante 42 dias de cultivo, o padrão de crescimento celular em ambas as culturas foi similar. O pH do meio de cultura BM0 e BM2 sofreu uma progressiva redução durante o período de cultivo. Em ambas as culturas embriogênicas foram observadas um consumo preferencial de glicose no período final da curva de crescimento celular. O nível de proteínas extracelulares foi similar para ambas as culturas embriogênicas. A dupla coloração com carmin acético e azul de Evans revelou diferenças na organização das linhagens celulares embriogênicas, sendo que a presença de proembriões somáticos bipolares foi apenas evidenciada nas culturas embriogênicas mantidas em meio de cultura líquido sem reguladores de crescimento.

  9. Tick cell culture isolation and growth of Rickettsia raoultii from Dutch Dermacentor reticulatus ticks. (United States)

    Alberdi, M Pilar; Nijhof, Ard M; Jongejan, Frans; Bell-Sakyi, Lesley


    Tick cell lines play an important role in research on ticks and tick-borne pathogenic and symbiotic microorganisms. In an attempt to derive continuous Dermacentor reticulatus cell lines, embryo-derived primary cell cultures were set up from eggs laid by field ticks originally collected as unfed adults in The Netherlands and maintained for up to 16 months. After several months, it became evident that cells in the primary cultures were infected with a Rickettsia-like intracellular organism. Supernatant medium containing some D. reticulatus cells was inoculated into cultures of 2 Rhipicephalus (Boophilus) microplus cell lines, BME/CTVM2 and BME/CTVM23, where abundant growth of the bacteria occurred intracellularly on transfer to both cell lines. Bacterial growth was monitored by light (live, inverted microscope, Giemsa-stained cytocentrifuge smears) and transmission electron microscopy revealing heavy infection with typical intracytoplasmic Rickettsia-like bacteria, not present in uninfected cultures. DNA was extracted from bacteria-infected and uninfected control cultures, and primers specific for Rickettsia 16S rRNA, ompB, and sca4 genes were used to generate PCR products that were subsequently sequenced. D. reticulatus primary cultures and both infected tick cell lines were positive for all 3 Rickettsia genes. Sequencing of PCR products revealed 99-100% identity with published Rickettsia raoultii sequences. The R. raoultii also grew abundantly in the D. nitens cell line ANE58, poorly in the D. albipictus cell line DALBE3, and not at all in the D. andersoni cell line DAE15. In conclusion, primary tick cell cultures and cell lines are useful systems for isolation and propagation of fastidious tick-borne microorganisms. In vitro isolation of R. raoultii from Dutch D. reticulatus confirms previous PCR-based detection in field ticks, and presence of the bacteria in the tick eggs used to initiate the primary cultures confirms that transovarial transmission of this

  10. The effect of culture conditions on the mycelial growth and luminescence of naturally bioluminescent fungi. (United States)

    Weitz, H J; Ballard, A L; Campbell, C D; Killham, K


    The effects of temperature, light and pH on mycelial growth and luminescence of four naturally bioluminescent fungi were investigated. Cultures of Armillaria mellea, Mycena citricolor, Omphalotus olearius and Panellus stipticus were grown at 5 degrees C, 15 degrees C, 22 degrees C and 30 degrees C, under 24 h light, 12 h light/12 h dark and 24 h dark, and at a pH ranging from 3.5 to 7 in three separate experiments. Temperature and pH had a significant effect on mycelial growth and bioluminescence, however light did not. Bioluminescence and mycelial growth were optimum at 22 degrees C and pH 3-3.5, the exception being M. citricolor for which bioluminescence and growth were optimum at pH 5-6 and pH 4, respectively. With the exception of M. citricolor, bioluminescence and mycelial growth were greater under 24 h darkness. An understanding of the effect of culture conditions on mycelial growth and luminescence is necessary for the future application of bioluminescent fungi as biosensors.

  11. A study on clavulanic acid production by Streptomyces clavuligerus in batch, fed-batch and continuous processes

    Directory of Open Access Journals (Sweden)

    A. B. Neto


    Full Text Available Clavulanic acid (CA is a potent inhibitor of beta-lactamases, enzymes that are responsible for the hydrolysis of beta-lactam antibiotics. It is a secondary metabolite produced by the filamentous aerobic bacterium Streptomyces clavuligerus in submerged cultivations. In the present work clavulanic acid production in batch, fed-batch and continuous bioreactors was studied with the objective of increasing productivity. The operating conditions: temperature, aeration and agitation, were the same in all cases, 28º C, 0.5 vvm and 800 rpm, respectively. The CA concentration obtained in the fed-batch culture, 404 mg L-1, was ca twice the value obtained in the batch culture, 194 mg L-1, while 293 mg L-1 was obtained in the continuous culture. The highest productivity was obtained in the continuous cultivation, 10.6 mg L-1 h-1, as compared with 8.8 mg L-1 h-1 in the fed-batch process and 3.5 mg L-1 h-1 in the batch process, suggesting that continuous culture of Streptomyces clavuligerus is a promising strategy for clavulanic acid production.

  12. Effects of in vitro growth culture duration and prematuration culture on maturational and developmental competences of bovine oocytes derived from early antral follicles. (United States)

    Huang, Weiping; Nagano, Masashi; Kang, Sung-Sik; Yanagawa, Yojiro; Takahashi, Yoshiyuki


    Bovine ovaries offer a large pool of oocytes that could be used for in vitro production of embryos of genetically valuable animals. The effects of in vitro growth (IVG) culture duration (10, 12, and 14 days) on the viability and growth of bovine oocytes derived from early antral follicles (0.5-1 mm diameter) in this study. In addition, the effect of pre-IVM culture with phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine) on nuclear maturation of IVG oocytes was also evaluated. In experiment 1, oocyte viability observed after 10 or 12 days of IVG culture was greater (P culture. Oocyte diameters and proportions of oocytes at metaphase II stage were greater (P culture where used when compared with 10 days culture. In addition, the proportion of oocytes at metaphase II stage was greater (P culture was performed for oocytes derived from 12 and 14 days of IVG culture. When 12 and 14 days of IVG culture followed by pre-IVM culture were compared in experiment 2, cumulus cell membrane integrity was greater (P culture (24.5%) was greater (P culture was considered the optimal processing system for bovine oocytes derived from early antral follicles when oocyte viability, diameter, maturation, and development competences were considered.

  13. Differences in stationary-phase cells of a commercial Saccharomyces cerevisiae wine yeast grown in aerobic and microaerophilic batch cultures assessed by electric particle analysis, light diffraction and flow cytometry. (United States)

    Portell, X; Ginovart, M; Carbó, R; Vives-Rego, J


    We applied electric particle analysis, light diffraction and flow cytometry to obtain information on the morphological changes during the stationary phase of Saccharomyces cerevisiae. The reported analyses of S. cerevisiae populations were obtained under two different conditions, aerobic and microaerophilic, at 27°C. The samples analysed were taken at between 20 and 50 h from the beginning of culture. To assist in the interpretation of the observed distributions a complexity index was used. The aerobically grown culture reached significantly greater cell density. Under these conditions, the cell density experienced a much lower reduction (3%) compared with the microaerophilic conditions (30%). Under aerobic conditions, the mean cell size determined by both electric particle analysis and light diffraction was lower and remained similar throughout the experiment. Under microaerophilic conditions, the mean cell size determined by electric particle analysis decreased slightly as the culture progressed through the stationary phase. Forward and side scatter distributions revealed two cell subpopulations under both growth conditions. However, in the aerobic growing culture the two subpopulations were more separated and hence easier to distinguish. The distributions obtained with the three experimental techniques were analysed using the complexity index. This analysis suggested that a complexity index is a good descriptor of the changes that take place in a yeast population in the stationary phase, and that it aids in the discussion and understanding of the implications of these distributions obtained by these experimental techniques.

  14. Effect of culture conditions on the growth of biomass Yarrowia lipolytica - producing protein feed

    Directory of Open Access Journals (Sweden)

    O. S. Korneeva


    Full Text Available Fodder yeast is highly valuable protein-vitamin products. Protein digestibility by yeast and amino acid content, superior proteins of animal origin. Fodder yeast protein digested in animals by 95 %. The biological value of yeast protein is determined by the presence of a significant amount of essential amino acids. Moreover, yeast cells contain many vitamins microelement and a significant amount of fat, in which the predominant unsaturated fatty acid. Currently, fodder yeast successfully used in livestock and poultry, so the demand for them is increasing every year. For the production of fodder yeast using a yeast having the necessary technological properties: the ability of rapid growth in aerobic conditions to form protein, amino acids and vitamins, resistant crop production, the development of resistance to foreign microorganisms. Intensive education yeast biomass contributes to a number of conditions, including pH, temperature and aeration of the culture occupy an important place. The main criterion for comparison and selection of a culture medium for this is the speed of its growth and ability to assimilate all of the nutrients with high economic factor. It depends on the performance of the enterprise, energy consumption and other technical - economic performance. The effect of pH of the medium on the biomass accumulation of yeast Yarrowia lipolytica. Found that at pH 5,2 - 5,5 observed maximum growth rate of the yeast cells. The effect of temperature on the accumulation of yeast biomass. The temperature of the culture medium determines the intensity of metabolism in cells. It was found that the optimal growth temperature of the culture Yarrowia lipolytica is 33 0C. The effect of aeration on the growth rate of yeast cells. Tro-established that the maximum increase of biomass was obtained with the aeration of 70 cm3 /cm3hrs.

  15. Comparative effects of contraction and angiotensin II on growth of adult feline cardiocytes in primary culture (United States)

    Wada, H.; Zile, M. R.; Ivester, C. T.; Cooper, G. 4th; McDermott, P. J.


    The purposes of this study were 1) to determine whether angiotensin II causes growth of adult feline cardiocytes in long-term culture, 2) to compare the growth effects of angiotensin II with those resulting from electrically stimulated contraction, and 3) to determine whether the anabolic effects of contraction are exerted via the angiotensin type 1 receptor. Adult feline cardiocytes were cultured on laminin-coated trays in a serum-free medium. Cardiocytes were either electrically stimulated to contract (1 Hz, 5-ms pulse duration, alternating polarity) or were nonstimulated and quiescent. Quiescent cells were studied as controls and after treatment with angiotensin II (10(-8) M), losartan (10(-6) M; an angiotensin type 1-receptor antagonist), or angiotensin II plus losartan. Contracting cells were studied in the presence and absence of angiotensin II or losartan. In quiescent cardiocytes, angiotensin II treatment on day 7 significantly increased protein synthesis rates by 22% and protein content per cell by 17%. The effects of angiotensin II were completely blocked by losartan. Electrically stimulated contraction on days 4 and 7 in culture significantly increased protein synthesis rate by 18 and 38% and protein content per cell by 19 and 46%, respectively. Angiotensin II treatment did not further increase protein synthesis rate or protein content in contracting cardiocytes. Furthermore, losartan did not block the anabolic effects of contraction on protein synthesis rates or protein content. In conclusion, angiotensin II can exert a modest anabolic effect on adult feline cardiocytes in culture. In contracting feline cardiocytes, angiotensin II has no effect on growth. Growth caused by electrically stimulated contraction occurs more rapidly and is greater in magnitude than that caused by angiotensin II. Growth of contracting adult feline cardiocytes is not dependent on activation of the angiotensin receptor.

  16. Metabolic Flux Analysis Model of Hybridoma Cell and Its Application to Batch Culture%杂交瘤细胞代谢流分析模型及其在批式培养中的应用

    Institute of Scientific and Technical Information of China (English)

    仝志明; 曹竹安


    An improved mathematical model of metabolic flux analysis bymaterial balances and linear programming method was established, which was first applied to batch culture process in this paper. The analysis results indicate that the metabolic efficiency of substrates such as glucose is very low. It showed that the response of cell metabolism to the step change of glutamine and other amino acid concentration was time-delayed. Metabolic efficiency was higher both in lag phase with highest substrate concentrations and glutamine-limited phase than that in exponential phase. It was also concluded that restraining glutamine from entering energy metabolism could reduce the accumulation of both ammonia and lactate.%通过物料衡算和线性规划方法建立了进行代谢流分析的数学模型,并首次应用于批式培养过程.分析结果表明,细胞对葡萄糖等底物的代谢效率很低,细胞对谷氨酰胺等氨基酸浓度的变化响应迟缓,在底物充裕的迟滞期及谷氨酰胺限制条件下的代谢效率高于对数生长期.另外限制谷氨酰胺等氨基酸进入能量代谢,既可减少氨的积累,也会减少乳酸的生成.

  17. Changes of cytokinin nucleotides in an anise cell culture (Pimpinella anisum L.) during growth and embryogenesis. (United States)

    Ernst, D; Oesterhelt, D


    Endogenous levels of cytokinin nucleotides in an anise cell culture were determined during proembryonal, as well as embryonal development. In both cultures the maximum level of isopentenyladenine nucleotides was found during the first four days of incubation which correlated with the beginning of logarithmic growth (embryonal: 8 ng g(-1) tresh weight; proembryonal: 17.4 ng g(-1) fresh weight). The concentration of zeatin nucleotides remained constant at a very low level. The present data and those of Ernst et al. (1984) and Ernst and Oesterhelt (1984) are concerned in ascribing a major role to cytokinins in cell division, but not in embryo differentiation.

  18. Expression of Transforming Growth Factor-β in Cultured Normal Human Lens Epithelia Cells

    Institute of Scientific and Technical Information of China (English)

    黄渝侃; 魏厚仁


    Summary: In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor β (TGF-β), reverse transcriptase polymerase chain reaction (RTPCR) and immunohistochemical methods were used for detection of TGF-β mRNA and protein in cultured normal human LEC. The results showed that a single RT-PCR amplified product about 310bp was obtained, and the sequence was homologous to the known sequence. TGF-β immunostain was positive in the plasma of LEC. It was suggested that normal human LEC could produce TGF-β, and LEC could be affected by TGF-β through autocrine action.

  19. Impact of different culture media on hairy roots growth of Valeriana officinalis L.


    Pakdin Parizi, Ali; Farsi, Mohammad; Nematzadeh, Ghorban-Ali; Amin MIRSHAMSI


    Transformed hairy root cultures of Valeriana officinalis were established by infection with Agrobacterium rhizogenes strain ATCC 15834. To determine the effect of different media on the growth of V. officinalis hairy roots, MS, B5 media (1.0X and 0.5X strength), N6 medium and a modified MS medium without phytohormones were used. In addition, different NH4+ to NO3- ratios in MS medium were studied. The effects of these treatments were evaluated after 21 days of culture in relation to hairy roo...

  20. Inhibitory Activity of Yokukansankachimpihange against Nerve Growth Factor-Induced Neurite Growth in Cultured Rat Dorsal Root Ganglion Neurons

    Directory of Open Access Journals (Sweden)

    Chiaki Murayama


    Full Text Available Chronic pruritus is a major and distressing symptom of many cutaneous diseases, however, the treatment remains a challenge in the clinic. The traditional Chinese-Japanese medicine (Kampo medicine is a conservative and increasingly popular approach to treat chronic pruritus for both patients and medical providers. Yokukansankachimpihange (YKH, a Kampo formula has been demonstrated to be effective in the treatment of itching of atopic dermatitis in Japan although its pharmacological mechanism is unknown clearly. In an attempt to clarify its pharmacological actions, in this study, we focused on the inhibitory activity of YKH against neurite growth induced with nerve growth factor (NGF in cultured rat dorsal root ganglion (DRG neurons because epidermal hyperinnervation is deeply related to itch sensitization. YKH showed approximately 200-fold inhibitory activity against NGF-induced neurite growth than that of neurotropin (positive control, a drug used clinically for treatment of chronic pruritus. Moreover, it also found that Uncaria hook, Bupleurum root and their chemical constituents rhynchophylline, hirsutine, and saikosaponin a, d showed inhibitory activities against NGF-induced neurite growth, suggesting they should mainly contribute to the inhibitory activity of YKH. Further study on the effects of YKH against epidermal nerve density in “itch-scratch” animal models is under investigation.

  1. Primary neuron culture for nerve growth and axon guidance studies in zebrafish (Danio rerio.

    Directory of Open Access Journals (Sweden)

    Zheyan Chen

    Full Text Available Zebrafish (Danio rerio is a widely used model organism in genetics and developmental biology research. Genetic screens have proven useful for studying embryonic development of the nervous system in vivo, but in vitro studies utilizing zebrafish have been limited. Here, we introduce a robust zebrafish primary neuron culture system for functional nerve growth and guidance assays. Distinct classes of central nervous system neurons from the spinal cord, hindbrain, forebrain, and retina from wild type zebrafish, and fluorescent motor neurons from transgenic reporter zebrafish lines, were dissociated and plated onto various biological and synthetic substrates to optimize conditions for axon outgrowth. Time-lapse microscopy revealed dynamically moving growth cones at the tips of extending axons. The mean rate of axon extension in vitro was 21.4±1.2 µm hr(-1 s.e.m. for spinal cord neurons, which corresponds to the typical ∼0.5 mm day(-1 growth rate of nerves in vivo. Fluorescence labeling and confocal microscopy demonstrated that bundled microtubules project along axons to the growth cone central domain, with filamentous actin enriched in the growth cone peripheral domain. Importantly, the growth cone surface membrane expresses receptors for chemotropic factors, as detected by immunofluorescence microscopy. Live-cell functional assays of axon extension and directional guidance demonstrated mammalian brain-derived neurotrophic factor (BDNF-dependent stimulation of outgrowth and growth cone chemoattraction, whereas mammalian myelin-associated glycoprotein inhibited outgrowth. High-resolution live-cell Ca(2+-imaging revealed local elevation of cytoplasmic Ca(2+ concentration in the growth cone induced by BDNF application. Moreover, BDNF-induced axon outgrowth, but not basal outgrowth, was blocked by treatments to suppress cytoplasmic Ca(2+ signals. Thus, this primary neuron culture model system may be useful for studies of neuronal development

  2. Nitrifying and denitrifying bacteria in aerobic granules formed in sequencing batch airlift reactors

    Institute of Scientific and Technical Information of China (English)

    WANG Fang; YANG Fenglin; QI Aijiu


    The purpose of this study was to investigate nitrifying bacteria and denitrifying bacteria isolated from aerobic granules.Aerobic granules were formed in an internal-circulate sequencing batch airlift reactor(SBAR)and biodegradation of NH3 -N was analyzed in the reactor.Bacteria were isolated and determined from aerobic granules using selected media.The growth properties and morphology of bacteria colonies were observed by controlling aerobic or anaerobic conditions in the culture medium.It was found that bacteria in aerobic granules were diverse and some of them were facultative aerobes.The diversity of bacteria in aerobic granules was a premise of simultaneous nitrification and denitrification.

  3. Vision Marker-Based In Situ Examination of Bacterial Growth in Liquid Culture Media (United States)

    Kim, Kyukwang; Choi, Duckyu; Lim, Hwijoon; Kim, Hyeongkeun; Jeon, Jessie S.


    The detection of bacterial growth in liquid media is an essential process in determining antibiotic susceptibility or the level of bacterial presence for clinical or research purposes. We have developed a system, which enables simplified and automated detection using a camera and a striped pattern marker. The quantification of bacterial growth is possible as the bacterial growth in the culturing vessel blurs the marker image, which is placed on the back of the vessel, and the blurring results in a decrease in the high-frequency spectrum region of the marker image. The experiment results show that the FFT (fast Fourier transform)-based growth detection method is robust to the variations in the type of bacterial carrier and vessels ranging from the culture tubes to the microfluidic devices. Moreover, the automated incubator and image acquisition system are developed to be used as a comprehensive in situ detection system. We expect that this result can be applied in the automation of biological experiments, such as the Antibiotics Susceptibility Test or toxicity measurement. Furthermore, the simple framework of the proposed growth measurement method may be further utilized as an effective and convenient method for building point-of-care devices for developing countries. PMID:27999349

  4. In vitro storage of cedar shoot cultures under minimal growth conditions. (United States)

    Renau-Morata, Begoña; Arrillaga, Isabel; Segura, Juan


    We developed procedures for slow-growth storage of Cedrus atlantica and Cedrus libani microcuttings of juvenile and adult origin, noting factors favouring the extension of subculture intervals. Microcuttings could be stored effectively up to 6 months at 4 degrees C and reduced light intensity, provided that they were grown on a diluted modified MS medium. The addition of 6% mannitol to the storage media affected negatively survival and multiplication capacity of the cultures. The slow-growth storage conditions used in our experiments did not induce remarkable effects on both RAPD variability and average DNA methylation in the species.

  5. Modulations of prolactin and growth hormone gene expression and chromatin structure in cultured rat pituitary cells.


    Levy-Wilson, B


    I have measured the effect of hormones and other regulatory factors present in the serum component of the culture medium on the levels of growth hormone and prolactin mRNAs in rat pituitary (GH4) cells. Hybridization of cytoplasmic RNA with growth hormone or prolactin cDNA clones indicate that serum depletion reduces significantly the amount of these two mRNAs. The localization of these two genes in chromatin was also analysed using micrococcal nuclease as a probe. At intermediate levels of d...

  6. Asymmetric cancer-cell filopodium growth induced by electric-fields in a microfluidic culture chip. (United States)

    Wang, Chun-Chieh; Kao, Yu-Chiu; Chi, Pei-Yin; Huang, Ching-Wen; Lin, Jiunn-Yuan; Chou, Chia-Fu; Cheng, Ji-Yen; Lee, Chau-Hwang


    We combine a micro-fluidic electric-field cell-culture (MEC) chip with structured-illumination nano-profilometry (SINAP) to quantitatively study the variations of cancer cell filopodia under external direct-current electric field (dcEF) stimulations. Because the lateral resolution of SINAP is better than 150 nm in bright-field image modality, filopodia with diameters smaller than 200 nm can be observed clearly without fluorescent labeling. In the MEC chip, a homogeneous EF is generated inside the culture area that simulates the endogenous EF environment. With this MEC chip-SINAP system, we directly observe and quantify the biased growth of filopodia of lung cancer cells toward the cathode. The epidermal growth factor receptors around the cell edges are also redistributed to the cathodal side. These results suggest that cancer-cell filopodia respond to the changes in EFs in the microenvironment.

  7. Endothelial cells stimulate growth of normal and cancerous breast epithelial cells in 3D culture

    Directory of Open Access Journals (Sweden)

    Magnusson Magnus K


    Full Text Available Abstract Background Epithelial-stromal interaction provides regulatory signals that maintain correct histoarchitecture and homeostasis in the normal breast and facilitates tumor progression in breast cancer. However, research on the regulatory role of the endothelial component in the normal and malignant breast gland has largely been neglected. The aim of the study was to investigate the effects of endothelial cells on growth and differentiation of human breast epithelial cells in a three-dimensional (3D co-culture assay. Methods Breast luminal and myoepithelial cells and endothelial cells were isolated from reduction mammoplasties. Primary cells and established normal and malignant breast cell lines were embedded in reconstituted basement membrane in direct co-culture with endothelial cells and by separation of Transwell filters. Morphogenic and phenotypic profiles of co-cultures was evaluated by phase contrast microscopy, immunostaining and confocal microscopy. Results In co-culture, endothelial cells stimulate proliferation of both luminal- and myoepithelial cells. Furthermore, endothelial cells induce a subpopulation of luminal epithelial cells to form large acini/ducts with a large and clear lumen. Endothelial cells also stimulate growth and cloning efficiency of normal and malignant breast epithelial cell lines. Transwell and gradient co-culture studies show that endothelial derived effects are mediated - at least partially - by soluble factors. Conclusion Breast endothelial cells - beside their role in transporting nutrients and oxygen to tissues - are vital component of the epithelial microenvironment in the breast and provide proliferative signals to the normal and malignant breast epithelium. These growth promoting effects of endothelial cells should be taken into consideration in breast cancer biology.

  8. Growth theory after Keynes, part II: 75 years of obstruction by the mainstream economics culture

    Directory of Open Access Journals (Sweden)

    Hendrik Van den Berg


    Full Text Available Part I of this essay explained the sequence of events that enabled the neoclassical paradigm to regain its dominant position in mainstream economics following serious challenges by ‘Keynesian’ economists. This second essay seeks to answer the question of why the economics profession was so willing to sustain the neoclassical paradigm in the face of the reality-based challenges by ‘Keynesian’ economists like Harrod and Domar. The answer is sought in the culture of economics, the history of science in general, and the study of power in the field of political economy. This article draws heavily on the work of the French sociologist Pierre Bourdieu, who divides culture into habitus (procedures and dispositions and doxa (more abstract beliefs and philosophies, in order to provide insight into how culture affects economic thinking. Bourdieu’s concept of symbolic violence helps to explain how a narrower neoclassical growth model was enthusiastically accepted as a replacement for the ‘Keynesian’ Harrod-Domar growth model. Financial and business interests clearly understood the power of culture and they used their accumulated wealth to support the neoliberal doxa and neoclassical habitus that would induce economists to willingly provide intellectual cover for policies that benefitted those financial and business interests. We conclude with a discussion on how the history of thought on economic development might have evolved if the Keynesian paradigm, and its dynamic Harrod-Domar model, had prevailed

  9. Some aspects of the growth and yield of Gracilaria tikvahiae in culture

    Energy Technology Data Exchange (ETDEWEB)

    Lapointe, B.E.; Ryther, J.H.


    A series of outdoor, continuous-flow seawater cultures (50 l; 0.23 m/sup 2/) were used to investigate the effects of culture density (kg/m/sup 2/), nutrient loading (total nitrogen input/day) with both NH/sub 4//sup +/-N and NO/sub 3//sup -/-N, and turnover rate (flow rate/culture volume) on the growth and yield of Gracilaria tikvahiae. Although specific growth rates as high as 60% per day were recorded for Gracilaria at low densities (0.4 kg wet wt/m/sup 2/) in summer conditions, maximum year-round yields were obtained at densities of 2.0 to 3.0 kg wet wt/m/sup 2/. Above a minimal daily nitrogen loading, the yield of Gracilaria was independent of (1) nutrient concentration, (2) nitrogen loading, or (3) whether nitrogen was in the form of NH/sub 4//sup +/-N or NO/sub 3//sup -/-N, but was (4) highly dependent upon flow rate. The time weighted mean annual production during 1976--1977 was 34.8 g dry wt/m/sup 2/.day or 127 t/ha.yr based on 12-months continuous operation at near optimal densities and flow rates in the non-nutrient limited culture system.

  10. Sterol and sesquiterpenoid biosynthesis during a growth cycle of tobacco cell suspension cultures. (United States)

    Chappell, J; Von Lanken, C; Vögeli, U; Bhatt, P


    The accumulation and biosynthesis of sterols and fungal elicitor-inducible sesquiterpenoids by tobacco (Nicotiana tabacum) cell suspension cultures were examined as a function of a 10 day culture cycle. Sterols accumulated concomitantly with fresh weight gain. The rate of sterol biosynthesis, measured as the incorporation rate of [(14)C]acetate and [(3)H]mevalonate, was maximal when the cultures entered into their rapid phase of growth. Changes in squalene synthetase enzyme activity correlated more closely with thein vivo synthesis rate and accumulation of sterols than 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) enzyme activity. Cell cultures entering into the rapid phase of growth also responded maximally to fungal elicitor as measured by the production of capsidiol, an extracellular sesquiterpenoid. However, the rate of sesquiterpenoid biosynthesis, measured as the incorporation rate of [(14)C]acetate and [(3)H]mevalonate, could not be correlated with elicitor-inducible HMGR or sesquiterpene cyclase enzyme activities, nor elicitor-suppressible squalene synthetase enzyme activity.

  11. Temperature calibration of lacustrine alkenones using in-situ sampling and growth cultures (United States)

    Huang, Y.; Toney, J. L.; Andersen, R.; Fritz, S. C.; Baker, P. A.; Grimm, E. C.; Theroux, S.; Amaral Zettler, L.; Nyren, P. E.


    Sedimentary alkenones have been found in an increasing number of lakes around the globe. Studies using molecular biological tools, however, indicate that haptophyte species that produce lacustrine alkenones differ from the oceanic species. In order to convert alkenone unsaturation ratios measured in sediments into temperature, it is necessary to obtain an accurate calibration for individual lakes. Using Lake George, North Dakota, U.S. as an example, we have carried out temperature calibrations by both in-situ water column sampling and culture growth experiments. In-situ measured lake water temperatures in the lake show a strong correlation with the alkenone unsaturation indices (r-squared = 0.82), indicating a rapid equilibrium of alkenone distributions with the lake water temperature in the water column. We applied the in-situ calibration to down-core measurements for Lake George and generated realistic temperature estimates for the past 8 kyr. Algal isolation and culture growth, on the other hand, reveal the presence of two different types of alkenone producing haptophytes. The species making a predominant C37:4 alkenone (species A) produced much greater concentrations of alkenones per unit volume than the species that produced a predominant C37:3 alkenone (species B). It is the first time that a haptophyte species (species A) making a predominant C37:4 alkenone is cultured successfully and now replicated at four different growth temperatures. The distribution of alkeones in Lake George sediments matches extremely well with the alkenones produced by species A, indicating species A is likely the producer for the alkenones in the sediments. The alkenone unsaturation ratio of alkenones produced by species A haptophyte shows a primary dependence on growth temperature as expected, but the slope of change appears to vary depending on the growth stages. The implications of our findings for paleoclimate reconstructions using lacustrine alkenones will be discussed.

  12. Optimal control of a fed-batch fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Dekkers, R.M.


    The common cultivation of bakers' yeast is an aerobic fed-batch fermentation under sugar-limited growth. The ultimate objective of on-line computer control is to optimize the process through maximizing the productivity of biomass formation while minimizing the consumption of raw materials for the product. Results obtained on the optimal control of a fed-batch fermentation are given. The aspects to be considered are instrumentation, state estimation, optimization and process control.

  13. Effects of transforming growth factor-beta on long-term human cord blood monocyte cultures

    Energy Technology Data Exchange (ETDEWEB)

    Orcel, P.; Bielakoff, J.; De Vernejoul, M.C. (INSERM U18, Hopital Lariboisiere, Paris (France))


    Transforming growth factor-beta (TGF-beta) modulates growth and differentiation in many cell types and is abundant in bone matrix. We recently showed that human cord blood monocytes cultured in the presence of 1,25(OH)2D3 acquire some features of osteoclast precursors. Since TGF-beta has been shown to influence bone resorption in organ culture, we have studied the effect of TGF-beta (1-1,000 pg/ml) on cord blood monocyte cultures. These cells were cultured on plastic substrate during 3 weeks in the presence of 20% horse serum and 10(-9) M 1,25(OH)2D3. TGF-beta, from a concentration of 10 pg/ml in the culture medium, decreased in a dose dependent manner the formation of multinucleated cells. At a concentration of TGF-beta of 1 ng/ml, the multinucleated cells were reduced to 2.1% +/- 0.3%, compared to 19.3% +/- 1.5% in control cultures. TGF-beta inhibited in a dose-dependent manner the proliferation of cord blood monocytes as assessed by 3H-thymidine incorporation at 7 and 14 days of culture. The fusion index was also decreased by 3 weeks of treatment with TGF-beta. Indomethacin did not reverse the inhibitory effects of TGF-beta. The expression of the osteoclastic phenotype was assessed using two different antibodies: 23C6, a monoclonal antibody directed against the vitronectin receptor, which is highly expressed by osteoclasts but not by adult monocytes, and an antibody to HLA-DR, which is not present on osteoclast. TGF-beta decreased the expression of HLA-DR and increased in a dose-dependent manner the proportion of 23C6-labeled cells; these results suggest that TGF-beta could modulate a differentiation effect to the osteoclastic phenotype. However, when cord blood monocytes were cultured on devitalized rat calvariae prelabeled with 45Ca, TGF-beta did not induce any 45Ca release from bone cultured with monocytes.

  14. Effect of radiation-degraded chitosan on growth promotion of flower plant in tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Luan, Le Quang; Ha, Vo Thi Thu; Hai, Le; Hien, Nguyen Quoc [Vietnam Atomic Energy Commission, Nuclear Research Institute, Dalat (Viet Nam); Nagasawa, Naotsugu; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment


    Radiation is a useful tool for degradation of polysaccharides, such as starch, carrageenan, alginate and chitin/chitosan. The viscosity molecular weight (Mw) of chitosan with 80% degree of deacetylation was reduced to 1.5 x 10{sup 5} by irradiation of 50kGy in solid phase. The solution of 10% of chitosan with Mw ca. 15 x 10{sup 5} was then irradiated at doses ranging 10-250kGy for further degradation and the products were supplemented into cultural media for testing of plant growth promotion effect. The results indicated that irradiated chitosan showed a strong growth-promotion effect on the increase of the length of shoot, the length of root and fresh biomass for flower plants namely Limonium latifolium, Eustoma grandiflorum and Chrysanthemum morifolium in tissue culture. The growth-promotion effect was obtained by the treatments with 50ppm of chitosan irradiated at the doses of 75-100kGy in 10% solution. The suitable concentrations of chitosan irradiated at 100kGy are ca. 100ppm for C. morifolium, 30ppm for E. grandiflorum and 40ppm for L. latifolium. In addition, our study also indicated that the survival ratio of transferred flower plantlets treated with irradiated chitosan was improved after acclimatizing for 30 days in the greenhouse. Accordingly, it is concluded that degraded chitosan obtained by radiation degradation technique is effective as a plant growth promoter as well as irradiated alginate. (author)

  15. Stimulation of Mucosal Mast Cell Growth in Normal and Nude Rat Bone Marrow Cultures (United States)

    Haig, David M.; McMenamin, Christine; Gunneberg, Christian; Woodbury, Richard; Jarrett, Ellen E. E.


    Mast cells with the morphological and biochemical properties of mucosal mast cells (MMC) appear and proliferate to form the predominant cell type in rat bone marrow cultures stimulated with factors from antigen- or mitogen-activated lymphocytes. Conditioned media causing a selective proliferation of MMC were derived from mesenteric lymph node cells of Nippostrongylus brasiliensis-infected rats restimulated in vitro with specific antigen or from normal or infected rat mesenteric lymph node cells stimulated with concanavalin A. MMC growth factor is not produced by T-cell-depleted mesenteric lymph node cells or by the mesenteric lymph node cells of athymic rats. By contrast, MMC precursors are present in the bone marrow of athymic rats and are normally receptive to the growth factor produced by the lymphocytes of thymus-intact rats. The thymus dependence of MMC hyperplasia is thus based on the requirement of a thymus-independent precursor for a T-cell-derived growth promoter.

  16. Influences of Plant Growth Regulators,Basal Media and Carbohydrate Levels on Cell Suspension Culture of Panax ginseng

    Institute of Scientific and Technical Information of China (English)

    TangWei; WuJiongyuan; 等


    A cell suspension culture of Panax ginseng which may be continuously subcultured has been established.Embryogenic callus derived from clutured young leaves was used to initiate the culture,Plant growth regulators,basal medium formula and carbohydrate levels were examined to determine their various effects on suspension culture cell growth and development ,The best selection of plant growth regulator,basal medium and carbohydrate level is 2mg/L 2,4-D:0.5mg/L KT,MS and 3% sucrose respectively.

  17. Glutamate enhances the expression of vascular endothelial growth factor in cultured SD rat astrocytes

    Institute of Scientific and Technical Information of China (English)

    Chong-xiao Liu; Yong Liu; Wei Shi; Xin-lin Chen; Xin-li Xiao; Ling-yu Zhao; Yu-mei Tian; Jun-feng Zhang


    Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups: control group (C), glutamate group (G), QA group (Q), DCG-IV group (D), L-AP4 group (L) and glutanmte-FMCPG gronp (G+M). Cells were cultured under nomoxic condition (95% air, 5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes, respectively. G+ M group was preincubated with lmM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group, L group and C group. Different from that in C group, the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile, the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes, which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.

  18. Steroid-regulated growth of DDT1MF-2 cells is profoundly influenced by culture conditions. (United States)

    Lamb, D J; Ray, M


    DDT1MF-2 cells provide an ideal model for studying tumor-growth-stimulation by steroids. These cells progress to a rapidly proliferating, androgen-independent state after prolonged culture without androgen. After brief culture in different lots of fetal bovine serum (FBS), some lots induced a permanent state of hormone-independence in cells that had been androgen-responsive. To test the hypothesis that factors influenced androgen-responsive growth even after removal of serum, hormone-responsive DDT1MF-2 cells (7000 cells/well) were plated in medium Dulbecco's Modified Eagle Medium/F-12 Nutrition Mixture (1:1)/1% ITS with (a) 0.1% FBS, (b) 0.1% NuSerum (c) 0.1% Hyclone, or (d) MCDB-110/0.1% ITS with 5 ng/ml bFGF. On Days 2-8, medium was replaced with D-MEM/F12/ITS with 10 nM testosterone (T), 10 nM triamcinolone acetonide (TA), or ethanol (control) and the cells counted. While testosterone induced a 1.4-fold increase in cell growth after exposure to FBS or NuSerum, maximal testosterone effect (3-6-fold increase) was observed after Hyclone. Hydroxyflutamide antagonized the fivefold increase in growth observed with testosterone, with a slight decrease of growth with cAMP for cells plated in Hyclone. Androgen-independent cells were unaffected by testosterone, hydroxyflutamide, or 8Br-cAMP [medium (a)]. Maximal inhibition by triamcinolone acetonide (0.25 of control) was observed with medium (d). The effect of testosterone and triamcinolone acetonide on secretion of mitogenic activity into conditioned medium was also evaluated. Although conditioned media from control and testosterone-treated cells were mitogenic in a dose-dependent manner, the media from cells treated with triamcinolone acetonide and testosterone+TA conditioned medium was not mitogenic--but, of note, it was not growth inhibitory.

  19. Modelling the effect of lactic acid bacteria from starter- and aroma culture on growth of Listeria monocytogenes in cottage cheese

    DEFF Research Database (Denmark)

    Østergaard, Nina Bjerre; Eklöw, Annelie; Dalgaard, Paw


    Four mathematical models were developed and validated for simultaneous growth of mesophilic lactic acid bacteria from added cultures and Listeria monocytogenes, during chilled storage of cottage cheese with freshor cultured cream dressing. The mathematical models include the effect of temperature...... cheese to improvemodel performance. The inhibiting effect of mesophilic lactic acid bacteria from added cultures on growth of L. monocytogenes was efficiently modelled using the Jameson approach. The new models appropriately predicted the maximum population density of L. monocytogenes in cottage cheese....... The developed models were successfully validated by using 25 growth rates for L. monocytogenes, 17 growth rates for lactic acid bacteria and a total of 26 growth curves for simultaneous growth of L. monocytogenes and lactic acid bacteria in cottage cheese. These data were used in combination with bias...

  20. Growth enhancement effect of BzATP on primary cultured astrocytes from rat brain

    Institute of Scientific and Technical Information of China (English)

    Hua-Zheng LIANG; Ying LIU; Zhu-Rong YE


    Objective To explore whether BzATP could promote the growth of primary cultured astrocytes (AS) of rat and its possible mechanism, and whether TGF-β1 was involved in the event. Methods The primary cultured AS were derived from new born Sprague-Dawley rats.Glial fibrillary acidic protein (GFAP) immunofluorescent stain was used to check the purity of cultured AS. Morphometry was used to detect the changes of AS. The proliferation index of AS was detected by BrdU incorporation assay. Western blot was used to detect the changes of GFAP under different conditions. Changes of TGF-β1 gene transcription were detected by RT-PCR. ELISA was utilized to detect the variation of TGF-β1 protein in the supernate. Results The purity of primary cultured AS reached to 99%. BzATP promoted the hypertrophy of AS including the elongation of AS processes and the enlargement of cell bodies, BzATP also promoted the expression of GFAP in existence of Ca2+, but had no effect on cell proliferation. BzATP increased the transcription of TGF-β1 mRNA and the release of TGF-β1 protein in existence of Ca2+. TGF-β1 neutralizing antibody partially inhibited the expression of GFAP induced by BzATP, but had no effect on AS proliferation and cell morphology. Conclusion BzATP enhanced the hypertrophy of primary cultured AS, increased the expression of GFAP partially through TGF-β1. Mechanisms of the enhancement of AS growth induced by BzATP other than TGF-51 pathway remains to be elucidated.

  1. Growth, cysts and kinetics of Borrelia garinii (Spirochaetales: Spirochaetacea in different culture media

    Directory of Open Access Journals (Sweden)

    Angela de Oliveira


    Full Text Available The aim of the present paper was to evaluate cyst formation and growth parameters of Borrelia garinii in a range of media differing in formulation and cost. A qualitative assessment of morphology and motility of B. garinii was conducted. All media were prepared aseptically and used in test tubes or Petri dishes. For each medium, the initial spirochete concentration was standardized to 10³ spirochets/mL. The following culture media were suitable to grow B. garinii: Barbour-Stoenner-Kelly, brain heart infusion and PMR. Growth was minimal at six weeks post-inoculation and maximum spirochete density was observed between 9-12 weeks. Often, the cultures developed cysts of different sizes, isolated or in groups, with a spiraled portion of variable sizes, mainly in unfavorable culture media. Brazilian Lyme disease-like illness, also known as Baggio-Yoshinari syndrome (BYS, is a new and interesting emerging tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, only during its cystic forms. It has been assumed that the peculiar clinical and laboratory features of BYS are consequential to the absence of a human sucker Ixodes ricinus complex tick at risk areas in Brazil, supporting the concept that the borrelia phenotypic expression pattern is modified as it is transmitted through the host.

  2. Optimization of culture conditions to obtain maximal growth of penicillin-resistant Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Rodriguez Carlos A


    Full Text Available Abstract Background Streptococcus pneumoniae, particularly penicillin-resistant strains (PRSP, constitute one of the most important causes of serious infections worldwide. It is a fastidious microorganism with exquisite nutritional and environmental requirements to grow, a characteristic that prevents the development of useful animal models to study the biology of the microorganism. This study was designed to determine optimal conditions for culture and growth of PRSP. Results We developed a simple and reproducible method for culture of diverse strains of PRSP representing several invasive serotypes of clinical and epidemiological importance in Colombia. Application of this 3-step culture protocol consistently produced more than 9 log10 CFU/ml of viable cells in the middle part of the logarithmic phase of their growth curve. Conclusion A controlled inoculum size grown in 3 successive steps in supplemented agar and broth under 5% CO2 atmosphere, with pH adjustment and specific incubation times, allowed production of great numbers of PRSP without untimely activation of autolysis mechanisms.

  3. Effects of nitrendipine on growth activity in cultured vascular smooth muscle cells. (United States)

    Absher, M P; Baldor, L; Warshaw, D M


    Proliferation and migration of smooth muscle cells (SMCs) in the arterial wall may play a role in the development of atherosclerosis and hypertension. If cell migration and proliferation are dependent on extracellular calcium, then treatment with calcium channel blockers such as nitrendipine may alter these cellular responses. In the studies reported here, proliferation and migration activities were assessed in cultured bovine carotid artery smooth muscle cells exposed to nitrendipine. SMCs in long-term culture are characterized by periods of either stable or enhanced proliferative activity. During the stable periods, 1 microM nitrendipine has no effect on proliferation, but during periods of enhanced proliferation, 1 microM nitrendipine augments growth by approximately 20%. SMC migration rates and interdivision times were determined from analysis of time-lapse cinematography films. During stable periods of growth, cell migration rate was inversely related to interdivision time (i.e., fast migrating cells had the shortest interdivision times). Treatment with 1 microM nitrendipine abolished the relationship between migration rate and interdivision time and prolonged interdivision times. These data suggest that the ability of nitrendipine to alter SMC proliferation, interdivision time, and migration is dependent upon the overall proliferative state of the culture.

  4. Experimental study on He- Ne laser irradiation to inhibit scar fibroblast growth in culture

    Institute of Scientific and Technical Information of China (English)

    舒彬; 吴宗耀; 郝林林; 曾登芬; 冯光锐; 林永辉


    To explore the inhibitory effect of He-Ne laser irradiation on fibroblast growth of hypertrophic scars in culture. Methods: He-Ne laser with wavelength of 632.8 nm,power density of 50 mW/cm2 and doses of 3 J/cm2,30 J/cm2, 90 J/cm2 and 180 J/cm2 was used to irradiate human scar fibroblasts in culture 1, 3 and 5 times respectively, and then the cell count and cell cycle analysis were done. Results: Repeated irradiation with He-Ne laser at dose of 180 J/cm2 three and five times led to an evident decrease in total cell number compared with that of the control group and there was a significant difference ( P <0.05). The cell cycle analysis showed after three and five times of irradiation with 180 J/cm2 He-Ne laser the cell number in S-phase decreased from 51% to 20% and 14% respectively, the cell number in G0/G1 phase increased from 28% to 55% and 60% respectively, and the cell percentage in Sub-G1 phase was 6.7% and 9.8% respectively. Conclusions: Repeated irradiation with 180 J/cm2 He-Ne laser can inhibit scar fibroblasts growth in culture.It may be that He-Ne laser irradiation causes cell stagnation in G0/G1 phase and apoptosis.

  5. Effects of inorganic lead on the differentiation and growth of cortical neurons in culture. (United States)

    Kern, M; Audesirk, T; Audesirk, G


    Lead exposure has devastating effects on the developing nervous system, producing morphological, cognitive, and behavioral deficits. To elucidate some of the mechanisms of lead neurotoxicity, we have examined its effects on the differentiation of several types of cultured neurons. Previously, we reported the effects of inorganic lead on several parameters of growth and differentiation of E18 rat hippocampal neurons and two types of neuroblastoma cells cultured in medium with 2% fetal calf serum (FCS) (Audesirk et al., 1991). In the present study, we report the effects of concentrations of lead ranging from 1nM to 1 mM on the differentiation of hippocampal neurons cultured in medium containing 10% FCS. In addition, we investigated lead effects on neurons isolated from the motor cortex region of the E18 rat embryo. Cortical neurons were exposed to lead in concentrations ranging from 0.1 nM to 1 mM in medium with either 10% FCS or 2% FCS for 48 hr. The effects of lead tended to be multimodal. Neurite initiation, which is highly sensitive to neurotoxic compounds, was inhibited by lead at both high and low concentrations, with no effects at intermediate levels. Medium with 10% FCS enhanced certain growth parameters and tended to reduce the effects of lead. There was an overall consistency in the effects of lead on motor cortex and hippocampal neurons.

  6. Using a medium of free amino acids to produce penicillin g acylase in fed-batch cultivations of Bacillus megaterium ATCC 14945

    Directory of Open Access Journals (Sweden)

    R. G. Silva


    Full Text Available The production of penicillin G acylase (PGA, an important industrial enzyme from a wild strain of Bacillus megaterium using a pool of free amino acids as substrate was studied in a bench-scale bioreactor. Experiments carried out in shakers showed that the substitution of casein for free amino acids in the presence of cheese whey was the culture medium that provided the highest productivity. Several cultivations were carried out in a bioreactor operated in either batch or fed-batch mode. Batch runs showed that enzyme production is associated with microorganism growth. The following set of amino acids was preferentially consumed: Ala, Arg, Asp, Gly, Lys, Ser, Thr and Trp. On the other hand, the rates of consumption of His, Ile, Leu, Met, Phe, Pro, Tyr and Val were lower.

  7. About the performance of Sphaerotilus natans to reduce hexavalent chromium in batch and continuous reactors

    Energy Technology Data Exchange (ETDEWEB)

    Caravelli, Alejandro H., E-mail: [Centro de Investigacion y Desarrollo en Criotecnologia de Alimentos (CIDCA), CCT - CONICET - La Plata. Fac., Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 La Plata (1900) (Argentina); Zaritzky, Noemi E., E-mail: [Centro de Investigacion y Desarrollo en Criotecnologia de Alimentos (CIDCA), CCT - CONICET - La Plata. Fac., Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 La Plata (1900) (Argentina); Fac. de Ingenieria, Universidad Nacional de La Plata, 48 y 115 La Plata (1900) (Argentina)


    The hexavalent chromium biological reduction constitutes a safe and economical detoxification procedure of wastewaters containing Cr(VI). However, little research has been done to evaluate Cr(VI) tolerance and reduction capacity of microbial cultures under different growth conditions. The aims of this work were (a) to evaluate the capacity of Sphaerotilus natans to reduce Cr(VI) to Cr(III) in a continuous system limited in carbon and energy source or in nitrogen source, (b) to evaluate the toxic effect of Cr(VI) on this microorganism, (c) to carry out a complete analysis of Cr(VI) reduction by S. natans not only in continuous regime but also in batch system, and (d) to model the obtained results mathematically. S. natans exhibited great resistance to Cr(VI) (19-78 mg l{sup -1}) and optimal growth in continuous and batch systems using a mineral medium supplemented only with citric acid as organic substrate. In carbon- and energy-limited continuous systems, a maximum percentual decrease in Cr(VI) by 13% was reached for low influent Cr(VI) concentration (4.3-5.32 mgCr(VI) l{sup -1}); the efficiency of the process did not notoriously increase as the length of cellular residence time was increased from 4.16 to 50 h. A nitrogen-limited continuous operation with a cellular residence time of 28.5 h resulted in a Cr(VI) decrease of approximately 26-32%. In batch system, a mathematical model allowed to predict the Cr(VI) concentration as a function of time and the ratio between the initial Cr(VI) concentration and that of the biomass. High concentrations of initial Cr(VI) and biomass produced the highest performance of the process of Cr(VI) reduction reached in batch system, aspects which should be considered in detoxification strategies of wastewaters.

  8. Maximum growth rate of Mycobacterium avium in continuous culture or chronically infected BALB/c mice. (United States)

    McCarthy, C M; Taylor, M A; Dennis, M W


    Mycobacterium avium is a human pathogen which may cause either chronic or disseminated disease and the organism exhibits a slow rate of growth. This study provides information on the growth rate of the organism in chronically infected mice and its maximal growth rate in vitro. M. avium was grown in continuous culture, limited for nitrogen with 0.5 mM ammonium chloride and dilution rates that ranged from 0.054 to 0.153 h-1. The steady-state concentration of ammonia nitrogen and M. avium cells for each dilution rate were determined. The bacterial saturation constant for growth-limiting ammonia was 0.29 mM (4 micrograms nitrogen/ml) and, from this, the maximal growth rate for M. avium was estimated to be 0.206 h-1 or a doubling time of 3.4 h. BALB/c mice were infected intravenously with 3 x 10(6) colony-forming units and a chronic infection resulted, typical of virulent M. avium strains. During a period of 3 months, the number of mycobacteria remained constant in the lungs, but increased 30-fold and 8,900-fold, respectively, in the spleen and mesenteric lymph nodes. The latter increase appeared to be due to proliferation in situ. The generation time of M. avium in the mesenteric lymph nodes was estimated to be 7 days.

  9. Growth inhibition of selected microorganisms by an association of dairy starter cultures and probiotics

    Directory of Open Access Journals (Sweden)

    Beniamino T. Cenci-Goga


    Full Text Available Several growth curves for selected pathogens and hygiene indicators alone and vs selected dairy starter cultures (LAB and commercial probiotics have been performed. All strains for LAB and commercial probiotics were inoculated as pure cultures into skim milk to get an initial cocci:bacilli:enterocci ratio of 2:1:1 and a concentration of approximately 107 cfu mL–1 until challenge vs selected pathogens and hygiene indicators. Selected pathogens came from the collection of the Laboratorio di Ispezione degli Alimenti di O.A. or were reference strains (Escherichia coli, CSH26 K12, Staphylococcus aureus 27R, Salmonella Derby 27, Pseudomonas fluorescens ATCC 13525, Listeria innocua ATCC 33090. Each strain was inoculated into skim milk to get an initial concentration of approximately 106 cfu mL–1. Growth curves in skim milk for the following challenges were studied: i sterility control; ii association LAB; iii association of LAB vs each selected pathogen or hygiene indicator; iv selected pathogen or hygiene indicator alone. The challenges were carried out in BHI broth and in skim milk at 37°C. The highest reduction was observed in milk but in general the association of LAB and the probiotic was able to limit the growth of pathogens and hygiene indicators.

  10. Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

    Directory of Open Access Journals (Sweden)

    Beatriz de Oliveira Costa


    Full Text Available The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL and Eutroferric Red Latosol (ERL soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05 in soils with glucose and inoculated with the fungi (except F. verticillioides, in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

  11. Data-driven batch schuduling

    Energy Technology Data Exchange (ETDEWEB)

    Bent, John [Los Alamos National Laboratory; Denehy, Tim [GOOGLE; Arpaci - Dusseau, Remzi [UNIV OF WISCONSIN; Livny, Miron [UNIV OF WISCONSIN; Arpaci - Dusseau, Andrea C [NON LANL


    In this paper, we develop data-driven strategies for batch computing schedulers. Current CPU-centric batch schedulers ignore the data needs within workloads and execute them by linking them transparently and directly to their needed data. When scheduled on remote computational resources, this elegant solution of direct data access can incur an order of magnitude performance penalty for data-intensive workloads. Adding data-awareness to batch schedulers allows a careful coordination of data and CPU allocation thereby reducing the cost of remote execution. We offer here new techniques by which batch schedulers can become data-driven. Such systems can use our analytical predictive models to select one of the four data-driven scheduling policies that we have created. Through simulation, we demonstrate the accuracy of our predictive models and show how they can reduce time to completion for some workloads by as much as 80%.

  12. Early stationary phase culture supernatant accelerates growth of sputum cultures collected after initiation of anti-tuberculosis treatment. (United States)

    Kolwijck, E; Friedrich, S O; Karinja, M N; van Ingen, J; Warren, R M; Diacon, A H


    We investigated the effect of Mycobacterium tuberculosis culture supernatant added to sputum cultures collected during the first 8 weeks of anti-tuberculosis treatment. With ongoing treatment duration, time to culture positivity decreased significantly in supernatant-enriched cultures, possibly due to stimulation of dormant or slowly metabolizing M. tuberculosis cells.

  13. Establishment and optimization of cell growth in suspension culture of Papaver bracteatum: a biotechnology approach for thebaine production


    FARJAMINEZHAD, Reza; Nasser ZARE; ASGHARI-ZAKARIA, Rasool; Farjaminezhad, Manoochehr


    Iranian poppy (Papaver bracteatum) is an important medicinal plant that is the main source of the opium alkaloids codeine, morphine, and thebaine. To establish an efficient protocol for cell suspension culture and growth, the effects of different plant growth regulators (2,4-D, NAA, BAP, and kinetin) on callus induction and cell suspension culture of Iranian poppy were evaluated. The maximum percentage of callus induction (86.67%) and fresh weight of callus were obtained in MS medium suppleme...

  14. Achyranthes bidentata Blume extract promotes neuronal growth in cultured embryonic rat hippocampal neurons

    Institute of Scientific and Technical Information of China (English)

    Xin Tang; Yiren Chen; Xiaosong Gu; Fei Ding


    We have prepared an aqueous extract of Achyranthes bidentata Blume,a commonly prescribed Chinese medicinal herb,and reported,in previous studies,that A.bidentata extract benefits nerve growth and prevents neuron apoptosis.In this study,we investigated the actions of ,4.bidentata extract on survival and growth of primarily cultured rat hippocampal neurons.The morphological observation revealed that neurite growth from hippocampal neurons was significantly enhanced by A.bidentata extract with similar effects to those induced by nerve growth factor (NGF),and the greatest neurite growth appeared on treatment with A.bidentata extract at 1 ttg/ml for 24 h.DNA microarray analysis indicated that there were 25 upregulated genes and 47 downregulated genes exhibiting significantly differential expression in hippocampal neurons treated with A.bidentata extract at 1 μg/ml for 6 h when compared to those in untreated hippocampal neurons.Real-time quantitative RT-PCR and Western blot analysis demonstrated that the expression of growth-associated protein-43 in hippocampal neurons was upregulated at both mRNA and protein levels after treatment with A.bidentata extract,and the optimal dosage of the extract was also 1 μg/ml.These data confirm that A.bidentata extract could promote in vitro hippocampal neuronal growth in a dose- and time-dependent manner.(C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences.Published by Elsevier Limited and Science in China Press.All rights reserved.

  15. Growth characteristics and functional changes in rat chondrocytes cultured in porous tantalum in vitro

    Directory of Open Access Journals (Sweden)

    Ling ZHANG


    Full Text Available Objective To evaluate the growth characteristics and functional changes in rat chondrocytes cultured in porous tantalum in vitro. Methods The chondrocytes isolated from cartilage of 3-week old SD rats were cultured in vitro, then the 2nd passage cells were identified and implanted in porous tantalum scaffolds with a density of 1×106 cells/ml. The morphological characteristics of the chondrocytes cultured in porous tantalum were observed under inverted microscope, scanning electron microscope (SEM and transmission electron microscope (TEM, and the content of glycosaminoglycan (GAG in the chondrocytes was measured by chromatometry. Results The harvested cells were identified as chondrocytes by type Ⅱ collagen immunocytochemical staining, toluidine blue staining and safranin-O staining. Many chondrocytes adhering to the edge of porous tantalum were found by inverted microscope. Observation under SEM showed that chondrocytes spread well on the surface and distributed in the holes of porous tantalum, and they proliferated and secreted some extracellular matrixes. TEM observation showed that the ultrastructure of chondrocytes cultured in porous tantalum was similar to that of normal chondrocytes. Chromatometry determination showed that the chondrocytes in porous tantalum could secrete GAG continuously. Conclusion Porous tantalum is shown to have a satisfactory biocompatibility with chondrocytes in vitro, and may be used as a scaffold for cartilage tissue engineering. DOI: 10.11855/j.issn.0577-7402.2014.06.08

  16. Analysis of single hyphal growth and fragmentation in submerged cultures using a population model

    DEFF Research Database (Denmark)

    Krabben, Preben; Nielsen, Søren; Michelsen, Michael Locht


    Carlo method is shown to be superior with respect to versatility. With measurements of 100 hyphal elements to represent a steady state and using the Monte Carlo method and a statistical test it is shown that one can discriminate between four models of hyphal fragmentation in submerged cultures. (C) 1997......Descriptions of population dynamics in submerged cultures are important when studying the mechanisms of growth and fragmentation of filamentous microorganisms. Population models are traditionally formulated as population balance equations. Population models of filamentous morphology are difficult...... to solve because of random fragmentation, which introduces an integral term into the population balance equations. Balances for the systemic properties, e.g. concentration of hyphal elements, substrate concentration, average total hyphal length, and average number of growing tips, are set up. Based...

  17. Inhibition of Fusarium graminearum growth in flour gel cultures by hexane-soluble compounds from oat (Avena sativa L.) flour. (United States)

    Doehlert, Douglas C; Rayas-Duarte, Patricia; McMullen, Michael S


    Fusarium head blight, incited by the fungus Fusarium graminearum, primarily affects wheat (Triticum aestivum) and barley (Hordeum vulgarum), while oat (Avena sativa) appears to be more resistant. Although this has generally been attributed to the open panicle of oats, we hypothesized that a chemical component of oats might contribute to this resistance. To test this hypothesis, we created culture media made of wheat, barley, and oat flour gels (6 g of flour in 20 ml of water, gelled by autoclaving) and inoculated these with plugs of F. graminearum from actively growing cultures. Fusarium growth was measured from the diameter of the fungal plaque. Plaque diameter was significantly smaller on oat flour cultures than on wheat or barley cultures after 40 to 80 h of growth. Ergosterol concentration was also significantly lower in oat cultures than in wheat cultures after growth. A hexane extract from oats added to wheat flour also inhibited Fusarium growth, and Fusarium grew better on hexane-defatted oat flour. The growth of Fusarium on oat flour was significantly and negatively affected by the oil concentration in the oat, in a linear relationship. A hexane-soluble chemical in oat flour appears to inhibit Fusarium growth and might contribute to oat's resistance to Fusarium head blight. Oxygenated fatty acids, including hydroxy, dihydroxy, and epoxy fatty acids, were identified in the hexane extracts and are likely candidates for causing the inhibition.

  18. Growth characteristics and nutrient depletion of Miscanthus x ogiformis Honda 'Giganteus' suspension cultures

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted


    The growth characteristics and nutrient depletion in suspension cultures of Miscanthus ogiformis Honda ‘Giganteus' grown in media containing either Murashige and Skoog or N6 basal nutrient salts were studied during a culture period of 15 days. Proline was added to both media in concentrations from...

  19. Growth and sporulation of Metarhizium flavoviride var. Flavoviride on culture media and lighting regimes

    Directory of Open Access Journals (Sweden)

    Onofre Sideney Becker


    Full Text Available Entomopathogenic fungi from the genus Metarhizium are largely used for the biological control of agricultural pests by conidia spreading on the field. Although conidia production is well studied in M. anisopliae, only few research studies were done in M. flavoviride. The present work was carried out alming to evaluate the Mycelial growth and sporulation of the entomopathogenic fungus Metarhizium flavoviride var. flavoviride growing at 27 ± 2°C on Potato-dextrose-agar (PDA, Czapek-agar (CZP and a complete agar medium (CM under three lighting regimes, (continuous illumination, light/dark cycle and an black light/dark cycle were investigated. A completely randomized 3 × 3 (culture media × lighting regime factorial design with four replicates was used. The best mycelial growth and sporulation occurred on the PDA and CM media under continuous illumination (P <= 0,05.

  20. Dupuytren's disease: comparative growth dynamics and morphology between cultured myofibroblasts (nodule) and fibroblasts (cord). (United States)

    Vande Berg, J S; Gelberman, R H; Rudolph, R; Johnson, D; Sicurello, P


    The excised palmar fascia of 11 patients with Dupuytren's disease was separated clinically into nodules and cords. Myofibroblasts were seen by light and electron microscopy in each of the nodules, but the cords generally lacked myofibroblasts. Only one cord specimen had microscopic features that were intermediate between nodule and cord. Electron microscopy demonstrated that in vivo differences between myofibroblasts from nodules and fibroblasts from cords and control skin samples could be preserved in vitro. Growth studies showed slower growth of cultured myofibroblasts (mean +/- SD generation time 68.7 +/- 15 h) than cord-derived fibroblasts (mean +/- SD generation time 51.5 +/- 0.9 h). These data suggest that the life cycle of the myofibroblasts from Dupuytren's disease nodules differs from that of fibroblasts found in cordlike tissues. These myofibroblasts have biological characteristics nearly identical to those of myofibroblasts found in other contracting tissues, such as granulating wounds and breast cancer.

  1. [Effects of bio-fertilizer on organically cultured cucumber growth and soil biological characteristics]. (United States)

    Cao, Dan; Zong, Liang-gang; Xiao, Jun; Zhang, Qian; Zhao, Yan


    Field trials of organic farming were conducted to examine the effects of different bio-fertilizers on the organically cultured cucumber growth, soil enzyme activities, and soil microbial biomass. Four treatments were installed, i. e., organic fertilizer only (CK), bio-fertilizer "Zhonghe" combined with organic fertilizer (ZHH), bio-fertilizer "NST" combined with organic fertilizer (NST), and bio-fertilizer "Bio" combined with organic fertilizer (BIO). Bio-fertilizers combined with organic fertilizer increased the cucumber yield significantly, and improved the root growth and leaf chlorophyll content. Comparing with that in CK, the cucumber yield in treatments ZHH, NST, and BIO was increased by 10.4%, 12.4%, and 29.2%, respectively. At the seedling stage, early flowering stage, and picking time of cucumber, the soil microbial biomass C and N in treatments ZHH, NST, and BIO were significantly higher than that in CK, and the activities of soil urease, acid phosphatase, and catalase were also higher.

  2. Effect of plant growth regulators on somatic embryogenesis in leaf cultures of Coffea canephora. (United States)

    Hatanaka, T; Arakawa, O; Yasuda, T; Uchida, N; Yamaguchi, T


    The effects of plant growth regulators on somatic embryogenesis were studied in leaf cultures of Coffea canephora. The maximum number of somatic embryos were obtained on media that contained only cytokinin as a plant growth regulator. All of the auxins tested (NAA, IBA, IAA and 2, 4-D) inhibited the formation of embryos. The optimal concentration of each cytokinin (2-iP, BA and kinetin) for somatic embryogenesis was 5 μM. Under optimal conditions, each explant formed more than 100 embryoids with little callus and few adventitious roots. Embryoids were formed only at the cut edges of the leaf discs. Cytokinins were absorbed only at the cut edges of leaf discs that were in contact with the medium, and were not transported to other parts of the explant.

  3. Bone morphogenetic protein-5 (BMP-5 promotes dendritic growth in cultured sympathetic neurons

    Directory of Open Access Journals (Sweden)

    Higgins Dennis


    Full Text Available Abstract Background BMP-5 is expressed in the nervous system throughout development and into adulthood. However its effects on neural tissues are not well defined. BMP-5 is a member of the 60A subgroup of BMPs, other members of which have been shown to stimulate dendritic growth in central and peripheral neurons. We therefore examined the possibility that BMP-5 similarly enhances dendritic growth in cultured sympathetic neurons. Results Sympathetic neurons cultured in the absence of serum or glial cells do not form dendrites; however, addition of BMP-5 causes these neurons to extend multiple dendritic processes, which is preceded by an increase in phosphorylation of the Smad-1 transcription factor. The dendrite-promoting activity of BMP-5 is significantly inhibited by the BMP antagonists noggin and follistatin and by a BMPR-IA-Fc chimeric protein. RT-PCR and immunocytochemical analyses indicate that BMP-5 mRNA and protein are expressed in the superior cervical ganglia (SCG during times of initial growth and rapid expansion of the dendritic arbor. Conclusions These data suggest a role for BMP-5 in regulating dendritic growth in sympathetic neurons. The signaling pathway that mediates the dendrite-promoting activity of BMP-5 may involve binding to BMPR-IA and activation of Smad-1, and relative levels of BMP antagonists such as noggin and follistatin may modulate BMP-5 signaling. Since BMP-5 is expressed at relatively high levels not only in the developing but also the adult nervous system, these findings suggest the possibility that BMP-5 regulates dendritic morphology not only in the developing, but also the adult nervous system.

  4. The thioamides methimazole and thiourea inhibit growth of M. avium Subspecies paratuberculosis in culture.

    Directory of Open Access Journals (Sweden)

    Robert J Greenstein

    Full Text Available BACKGROUND: Thyrotoxicosis is conceptualized as an "autoimmune" disease with no accepted infectious etiology. There are increasingly compelling data that another "autoimmune" affliction, Crohn disease, may be caused by Mycobacterium avium subspecies paratuberculosis (MAP. Like M. tb, MAP is systemic. We hypothesized that some cases of thyrotoxicosis may be initiated by a MAP infection. Because other thioamides treat tuberculosis, leprosy and M. avium complex, we hypothesized that a mode of action of some thioamide anti-thyrotoxicosis medications may include MAP growth inhibition. METHODS: The effect of the thioamides, thiourea, methimazole and 6-propo-2-thiouracil (6-PTU were studied in radiometric Bactec culture, on ten strains of three mycobacterial species (six of MAP, two of M. avium and two of M. tb. complex. Data are presented as "cumulative growth index," (cGI or "percent decrease in cumulative GI" (%-DeltacGI. PRINCIPAL FINDINGS: Methimazole was the most effective thioamide at inhibiting MAP growth. At 128microg/ml: MAP UCF-4; 65%-DeltacGI & MAP ATCC 19698; 90%-DeltacGI. Thiourea inhibited MAP "Ben" maximally; 70%-DeltacGI. Neither methimazole nor thiourea inhibited M. avium or M. tb. at the doses tested. 6-PTU has no inhibition on any strain studied, although a structurally analogous control, 5-PTU, was the most inhibitory thioamide tested. SIGNIFICANCE: We show inhibition of MAP growth by the thioamides, thiourea and methimazole in culture. These data are compatible with the hypothesis that these thioamides may have anti-prokaryotic in addition to their well-established eukaryotic actions in thyrotoxic individuals.

  5. Aligned Fibrous Scaffold Induced Aligned Growth of Corneal Stroma Cells in vitro Culture

    Institute of Scientific and Technical Information of China (English)

    GAO Yan; YAN Jing; CUI Xue-jun; WANG Hong-yan; WANG Qing


    To investigate the contribution of fibre arrangement to guiding the aligned growth of corneal stroma cells,aligned and randomly oriented fibrous scaffolds of gelatin and poly-L-lactic acid(PLLA) were fabricated by electrospinning.A comparative study of two different systems with corneal stroma cells on randomly organized and aligned fibres were conducted.The efficiency of the scaffolds for inducing the aligned growth of cells was assessed by morphological observation and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT) assay.Results show that the cells cultured on both randomly oriented and aligned scaffolds maintained normal morphology and well spreading as well as long term proliferation.Importantly,corneal stroma cells grew high orderly on the aligned scaffold,while the cells grew disordered on the randomly oriented scaffold.Moreover,the cells exhibited higher viability in aligned scaffold than that in randomly oriented scaffold.These results indcate that electrospinng to prepare aligned fibrous scaffolds has provided an effective approach to the aligned growth of corneal stroma cells in vitro.Our findings that fiber arrangement plays a crucial role in guiding the aligned growth of cells may be helpful to the development of better biomaterials for tissue engineered cornea.

  6. Growth performance of three pejerrey genetic groups in intensive culture system

    Directory of Open Access Journals (Sweden)

    Rafael Aldrighi Tavares


    Full Text Available This study examined the growth performance of Odontesthes bonariensis, O. humensis and hybrid (O. bonariensis X O. humensis in intensive culture system for a period of 180 days, using the body weight (W, total length (Lt, condition factor (CF, allometric coefficient (b, specific growth rate (SGR and estimates of body weight and maximum total length (Wmax, Ltmax as indicators. Fish were randomly selected from three genetic groups (Bonariensis, Humensis and Hybrid, divided into water recirculation system consisting of 12 tanks (50 L. 12 measurements of body weight and total length (0, 2, 7, 14, 21, 30, 45, 60, 75, 90, 120, 180 days were taken by means of a random sample of 10 subjects per replicate. All genetic groups grew in weight and length over the period analyzed, being influenced by the genetics of each species. O. bonariensis showed the best performance in growth and hybrid fish showed intermediate growth and estimated maximum body weight exceeding the pure species. Based on these results, it would be helpful the selective breeding and formation of domesticated strains.

  7. Red ginseng extract promotes the hair growth in cultured human hair follicles. (United States)

    Park, Gyeong-Hun; Park, Ki-young; Cho, Hong-il; Lee, Sang-Min; Han, Ji Su; Won, Chong Hyun; Chang, Sung Eun; Lee, Mi Woo; Choi, Jee Ho; Moon, Kee Chan; Shin, Hyoseung; Kang, Yong Jung; Lee, Dong Hun


    Ginseng has been shown to promote hair growth in several recent studies. However, its effects on human hair follicles and its mechanisms of action have not been sufficiently elucidated. This study aimed to investigate the hair growth-promoting effects of red ginseng extract (RGE) and its ginsenosides. The proliferative activities of cultured human hair follicles treated with RGE and ginsenoside-Rb1 were assessed using Ki-67 immunostaining. Their effects on isolated human dermal papilla cells (hDPCs) were evaluated using cytotoxicity assays, immunoblot analysis of signaling proteins, and the determination of associated growth factors. We examined the ability of RGE and ginsenosides to protect hair matrix keratinocyte proliferation against dihydrotestosterone (DHT)-induced suppression and their effects on the expression of androgen receptor. The in vivo hair growth-promoting effect of RGE was also investigated in C57BL/6 mice. Both RGE and ginsenoside-Rb1 enhanced the proliferation of hair matrix keratinocytes. hDPCs treated with RGE or ginsenoside-Rb1 exhibited substantial cell proliferation and the associated phosphorylation of ERK and AKT. Moreover, RGE, ginsenoside-Rb1, and ginsenoside-Rg3 abrogated the DHT-induced suppression of hair matrix keratinocyte proliferation and the DHT-induced upregulation of the mRNA expression of androgen receptor in hDPCs. Murine experiments revealed that the subcutaneous injection of 3% RGE resulted in more rapid hair growth than the negative control. In conclusion, RGE and its ginsenosides may enhance hDPC proliferation, activate ERK and AKT signaling pathways in hDPCs, upregulate hair matrix keratinocyte proliferation, and inhibit the DHT-induced androgen receptor transcription. These results suggest that red ginseng may promote hair growth in humans.

  8. BatchJobs and BatchExperiments: Abstraction Mechanisms for Using R in Batch Environments

    Directory of Open Access Journals (Sweden)

    Bernd Bischl


    Full Text Available Empirical analysis of statistical algorithms often demands time-consuming experiments. We present two R packages which greatly simplify working in batch computing environments. The package BatchJobs implements the basic objects and procedures to control any batch cluster from within R. It is structured around cluster versions of the well-known higher order functions Map, Reduce and Filter from functional programming. Computations are performed asynchronously and all job states are persistently stored in a database, which can be queried at any point in time. The second package, BatchExperiments, is tailored for the still very general scenario of analyzing arbitrary algorithms on problem instances. It extends package BatchJobs by letting the user define an array of jobs of the kind apply algorithm A to problem instance P and store results. It is possible to associate statistical designs with parameters of problems and algorithms and therefore to systematically study their influence on the results. The packages main features are: (a Convenient usage: All relevant batch system operations are either handled internally or mapped to simple R functions. (b Portability: Both packages use a clear and well-defined interface to the batch system which makes them applicable in most high-performance computing environments. (c Reproducibility: Every computational part has an associated seed to ensure reproducibility even when the underlying batch system changes. (d Abstraction and good software design: The code layers for algorithms, experiment definitions and execution are cleanly separated and enable the writing of readable and maintainable code.

  9. Influence of feeding conditions on clavulanic acid production in fed-batch cultivation with medium containing glycerol. (United States)

    Teodoro, Juliana C; Baptista-Neto, Alvaro; Cruz-Hernández, Isara L; Hokka, Carlos O; Badino, Alberto C


    First, the effect of different levels of nitrogen source on clavulanic acid (CA) production was evaluated in batch cultivations utilizing complex culture medium containing glycerol and three different levels of soy protein isolate (SPI). Cellular growth, evaluated in terms of the rheological parameter K, was highest with a SPI concentration of 30 g.L(-1) (4.42 g.L(-1) N total). However, the highest production of CA (380 mg.L(-1)) was obtained when an intermediate concentration of 20 g.L(-1) of SPI (2.95 g.L(-1) total N) was used. To address this, the influences of volumetric flow rate (F) and glycerol concentration in the complex feed medium (Cs(F)) in fed-batch cultivations were investigated. The best experimental condition for CA production was F=0.01 L.h(-1) and Cs(F)=120 g.L(-1), and under these conditions maximum CA production was practically twice that obtained in the batch cultivation. A single empirical equation was proposed to relate maximum CA production with F and Cs(F) in fed-batch experiments.

  10. Effects of Culture Conditions on Growth and Docosahexaenoic Acid Production from Schizochytrium limacinum

    Institute of Scientific and Technical Information of China (English)


    The effects of temperature, initial pH, salinity of culture medium, and carbon and nitrogen sources on growth and docosahexaenoic acid (C22: 6 n-3, DHA) production from Schizochytrium limacinum OUC88 were investigated in the present study. The results revealed that the optimal temperature, initial pH and salinity level of the medium for DHA production were 23 ℃, 7.0 and 18,respectively. Glucose was proved the best carbon source for the growth and DHA production from S. limacinum. Among the nitrogen sources tested, soybean cake hydrolysate, a cheap by-product, was found to be effective for the accumulation of DHA in S. limacinum cells. In addition, increasing the concentration of carbon sources in the medium caused a significant increase in cell biomass;however, accumulation of DHA in cells was mainly stimulated by the ratio of C/N in the medium. Under the optimal culture conditions, the maximum DHA yield achieved in flasks was 4.08 g L-1 after 5 d of cultivation.

  11. ARPE-19 cell growth and cell functions in euglycemic culture media. (United States)

    Heimsath, Ernest G; Unda, Richard; Vidro, Eileen; Muniz, Albert; Villazana-Espinoza, Elia T; Tsin, Andrew


    Human retinal pigmented epithelial cells (ARPE-19) grown in euglycemic media (5.5 mM) had lower cell number, significantly different cell morphology, and lower levels of vascular endothelial growth factor (VEGF) in the culture media than those grown in hyperglycemic media (18 mM) customarily used for culturing ARPE-19 cells. Although it has been shown that within a 24-hour period, all-trans retinoic acid significantly reduces VEGF secretion by retinal pigmented epithelial cells (grown in 18 mM glucose), such an inhibitory effect was not observed in cells grown in 5.5 mM glucose. Our results suggest that ARPE-19 cells grown in euglycemic media exhibit distinctly different cell growth, cell differentiation, and cell functions in comparison to cells grown in hyperglycemic media. Because euglycemic conditions provide a physiological glucose environment, this glucose concentration must be included in all future investigations of the mechanism of diabetic retinopathy when studying VEGF secretion by ARPE-19 cells.

  12. Expression of a synthetic gene encoding human insulin-like growth factor I in cultured mouse fibroblasts.


    Bayne, M L; Cascieri, M A; Kelder, B; Applebaum, J; Chicchi, G; Shapiro, J A; Pasleau, F.; Kopchick, J. J.


    A synthetic gene encoding human insulin-like growth factor I (hIGF-I) was assembled and inserted into an expression vector containing the cytomegalovirus immediate early (CMV-IE) transcriptional regulatory region and portions of the bovine growth hormone gene. The recombinant plasmid encodes a 97 amino acid fusion protein containing the first 27 amino acids of the bovine growth hormone precursor and the 70 amino acids of hIGF-I. This plasmid, when transiently introduced into cultured mouse fi...

  13. Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base Controlled Ecological Life Support System (CELSS) (United States)

    Venketeswaran, S.


    Experiments to determine whether plant tissue cultures can be grown in the presence of simulated lunar soil (SLS) and the effect of simulated lunar soil on the growth and morphogenesis of such cultures, as well as the effect upon the germination of seeds and the development of seedlings were carried out . Preliminary results on seed germination and seedling growth of rice and calli growth of winged bean and soybean indicate that there is no toxicity or inhibition caused by SLS. SLS can be used as a support medium with supplements of certain major and micro elements.

  14. Alginate as a cell culture substrate for growth and differentiation of human retinal pigment epithelial cells. (United States)

    Heidari, Razeih; Soheili, Zahra-Soheila; Samiei, Shahram; Ahmadieh, Hamid; Davari, Maliheh; Nazemroaya, Fatemeh; Bagheri, Abouzar; Deezagi, Abdolkhalegh


    The purpose of this study was to evaluate retinal pigment epithelium (RPE) cells' behavior in alginate beads that establish 3D environment for cellular growth and mimic extracellular matrix versus the conventional 2D monolayer culture. RPE cells were encapsulated in alginate beads by dripping alginate cell suspension into CaCl2 solution. Beads were suspended in three different media including Dulbecco's modified Eagle's medium (DMEM)/F12 alone, DMEM/F12 supplemented with 10 % fetal bovine serum (FBS), and DMEM/F12 supplemented with 30 % human amniotic fluid (HAF). RPE cells were cultivated on polystyrene under the same conditions as controls. Cell phenotype, cell proliferation, cell death, and MTT assay, immunocytochemistry, and real-time RT-PCR were performed to evaluate the effect of alginate on RPE cells characteristics and integrity. RPE cells can survive and proliferate in alginate matrixes. Immunocytochemistry analysis exhibited Nestin, RPE65, and cytokeratin expressions in a reasonable number of cultured cells in alginate beads. Real-time PCR data demonstrated high levels of Nestin, CHX10, RPE65, and tyrosinase gene expressions in RPE cells immobilized in alginate when compared to 2D monolayer culture systems. The results suggest that alginate can be used as a reliable scaffold for maintenance of RPE cells' integrity and in vitro propagation of human retinal progenitor cells for cell replacement therapies in retinal diseases.

  15. Roles of interleukin-9 in the growth and cholecystokinin-induced intracellular calcium signaling of cultured interstitial cells of Cajal. (United States)

    Gong, Yaoyao; Huang, Lei; Cheng, Wenfang; Li, Xueliang; Lu, Jia; Lin, Lin; Si, Xinmin


    Interstitial cells of Cajal (ICC) are pacemaker cells in the gastrointestinal (GI) tract and loss of ICC is associated with many GI motility disorders. Previous studies have shown that ICC have the capacity to regenerate or restore, and several growth factors are critical to their growth, maintenance or regeneration. The present study aimed to investigate the roles of interleukin-9 (IL-9) in the growth, maintenance and pacemaker functions of cultured ICC. Here, we report that IL-9 promotes proliferation of ICC, and culturing ICC with IL-9 enhances cholecystokinin-8-induced Ca²⁺ transients, which is probably caused by facilitating maintenance of ICC functions under culture condition. We also show co-localizations of cholecystokinin-1 receptor and IL-9 receptor with c-kit by double-immunohistochemical labeling. In conclusion, IL-9 can promote ICC growth and help maintain ICC functions; IL-9 probably performs its functions via IL-9 receptors on ICC.

  16. Effects of medium nutrition on cell growth and isocamptothecin A and B production by suspension cell culture of Camptotheca acuminata

    Institute of Scientific and Technical Information of China (English)

    Zhang Dongyan; Yu Fang; Bai Fengwu; An Lijia


    The effects of initial sucrose concentration, nitrate to ammonium ratio, total N concentration and phosphate concentration in medium on cell growth and isocamptothecin A and B synthesis by suspension cell culture of Camptotheca acuminata were investigated in 250 mL shake flasks. 30 g L-1 sucrose concentration was beneficial to secondary metabolites synthesis. The cell growth and metabolites synthesis were also affected by the ratio of NO-3/NH+4, and nitrate was favourable for cell growth. The maximum dry weight was achieved when nitrate was used as the sole N source. The effect of total initial N on the cell cultures was also investigated with NO-3/NH+4 ratio of 1∶2. The final dry cell weight was similar throughout culture period and 50 mM initial N was favourable for secondary metabolite synthesis. 50 mM initial phosphate concentration facilitated both cell growth and secondary metabolites synthesis.

  17. Roles of interleukin-9 in the growth and cholecystokinin-induced intracellular calcium signaling of cultured interstitial cells of Cajal.

    Directory of Open Access Journals (Sweden)

    Yaoyao Gong

    Full Text Available Interstitial cells of Cajal (ICC are pacemaker cells in the gastrointestinal (GI tract and loss of ICC is associated with many GI motility disorders. Previous studies have shown that ICC have the capacity to regenerate or restore, and several growth factors are critical to their growth, maintenance or regeneration. The present study aimed to investigate the roles of interleukin-9 (IL-9 in the growth, maintenance and pacemaker functions of cultured ICC. Here, we report that IL-9 promotes proliferation of ICC, and culturing ICC with IL-9 enhances cholecystokinin-8-induced Ca²⁺ transients, which is probably caused by facilitating maintenance of ICC functions under culture condition. We also show co-localizations of cholecystokinin-1 receptor and IL-9 receptor with c-kit by double-immunohistochemical labeling. In conclusion, IL-9 can promote ICC growth and help maintain ICC functions; IL-9 probably performs its functions via IL-9 receptors on ICC.

  18. Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

    Institute of Scientific and Technical Information of China (English)

    Zong-ming Wan; Lu Liu; Jian-yu Li; Rui-xin Li; Yong Guo; Hao Li; Jian-ming Zhang; Xi-zheng Zhang


    Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro. Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection. Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

  19. Type I TARPs promote dendritic growth of early postnatal neocortical pyramidal cells in organotypic cultures. (United States)

    Hamad, Mohammad I K; Jack, Alexander; Klatt, Oliver; Lorkowski, Markus; Strasdeit, Tobias; Kott, Sabine; Sager, Charlotte; Hollmann, Michael; Wahle, Petra


    The ionotropic α-amino-3-hydroxy-5-methyl-4-isoxazole propionate glutamate receptors (AMPARs) have been implicated in the establishment of dendritic architecture. The transmembrane AMPA receptor regulatory proteins (TARPs) regulate AMPAR function and trafficking into synaptic membranes. In the current study, we employ type I and type II TARPs to modulate expression levels and function of endogenous AMPARs and investigate in organotypic cultures (OTCs) of rat occipital cortex whether this influences neuronal differentiation. Our results show that in early development [5-10 days in vitro (DIV)] only the type I TARP γ-8 promotes pyramidal cell dendritic growth by increasing spontaneous calcium amplitude and GluA2/3 expression in soma and dendrites. Later in development (10-15 DIV), the type I TARPs γ-2, γ-3 and γ-8 promote dendritic growth, whereas γ-4 reduced dendritic growth. The type II TARPs failed to alter dendritic morphology. The TARP-induced dendritic growth was restricted to the apical dendrites of pyramidal cells and it did not affect interneurons. Moreover, we studied the effects of short hairpin RNA-induced knockdown of endogenous γ-8 and showed a reduction of dendritic complexity and amplitudes of spontaneous calcium transients. In addition, the cytoplasmic tail (CT) of γ-8 was required for dendritic growth. Single-cell calcium imaging showed that the γ-8 CT domain increases amplitude but not frequency of calcium transients, suggesting a regulatory mechanism involving the γ-8 CT domain in the postsynaptic compartment. Indeed, the effect of γ-8 overexpression was reversed by APV, indicating a contribution of NMDA receptors. Our results suggest that selected type I TARPs influence activity-dependent dendritogenesis of immature pyramidal neurons.

  20. Transforming growth factor-beta as a differentiating factor for cultured smooth muscle cells. (United States)

    Gawaziuk, J P; X; Sheikh, F; Cheng, Z-Q; Cattini, P A; Stephens, N L


    The aim of the present study was to determine whether the development of supercontractile smooth muscle cells, contributing to the nonspecific hyperreactivity of airways in asthmatic patients, is due to transforming growth factor (TGF)-beta. In cultured smooth muscle cells starved by removal of 10% foetal bovine serum for 7 days, growth arrest was seen; 30% became elongated and demonstrated super contractility. Study of conditioned medium suggested that the differentiating factor was TGF-beta. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was carried out on conditioned medium from the arrested cells. Two protein bands were identified as matrix metalloproteinase (MMP)-2 and TGF-beta1. To determine second messenger signalling by SMAD2, Western blotting and confocal microscopy were employed. Conditioned medium from arrested cultures showed the presence of MMP-2 and TGF-beta1, as revealed by SDS-PAGE; 68- and 25-kDa bands were seen. Differentiation was confirmed by upregulation of marker proteins, smooth muscle type myosin heavy chain and myosin light chain kinase. Confirmation was obtained by downregulating these proteins with decorin treatment, which reduces the levels of active TGF-beta and an adenoviral dominant-negative vector coding for a mutated type II TGF-beta-receptor. Activation of second messenger signalling was demonstrated immunocytochemically by the presence of phosphorylated SMAD2 and SMAD4. Transforming growth factor-beta is likely to be the differentiating factor responsible for the development of these supercontractile smooth muscle cells. The development of such cells in vivo after cessation of an asthmatic attack could contribute to the nonspecific hyperreactivity of airways seen in patients.

  1. Pure culture of Metarhizium anisopliae LHL07 reprograms soybean to higher growth and mitigates salt stress. (United States)

    Khan, Abdul Latif; Hamayun, Muhammad; Khan, Sumera Afzal; Kang, Sang-Mo; Shinwari, Zabta Khan; Kamran, Muhammad; Ur Rehman, Shafiq; Kim, Jong-Guk; Lee, In-Jung


    Little is known about the role of endophytic fungi against abiotic stresses and isoflavonoids (IF) contents of soybean. In current study, we investigated the role of fungal endophytes on the growth of soybean under salt stress conditions. Pure cultures of nine endophytic fungi were isolated from the roots of field-grown soybean plants, and their culture filtrates were screened on Waito-C and Dongjin-byeo rice cultivars; for identification of plant growth promoting fungal strains. It was observed that fungal isolate GMC-2B significantly promoted the growth of both Waito-C and Dongjin-byeo. GMC-2B was later identified as a new strain of Metarhizium anisopliae LHL07 on the basis of 18S rDNA sequences and phylogenetic analysis. Metarhizium anisopliae LHL07 inoculated soybean plants recorded significantly higher shoot length, shoot fresh and dry biomass, chlorophyll contents, transpiration rate, photosynthetic rate and leaf area; under sodium chloride induced salt stress as compared to non-inoculated control plants. An elevated proline and reduced superoxide dismutase and malondialdehyde contents in M. anisopliae LHL07 inoculated soybean plants demonstrated mitigation of salt induced oxidative stress. Furthermore, reduced abscisic acid and elevated jasmonic acid contents in soybean plants confirmed that lesser stress was convened to M. anisopliae inoculated-plants under salinity stress. We also assessed the role of M. anisopliae interaction on IF biosynthesis of soybean, and found significantly higher IF contents in M. anisopliae inoculated soybean plants. In conclusion, endophytic fungal interactions with soybean can be beneficial to improve soybean quality and quantity under salt affected agricultural systems.

  2. Modelling the effect of lactic acid bacteria from starter- and aroma culture on growth of Listeria monocytogenes in cottage cheese. (United States)

    Østergaard, Nina Bjerre; Eklöw, Annelie; Dalgaard, Paw


    Four mathematical models were developed and validated for simultaneous growth of mesophilic lactic acid bacteria from added cultures and Listeria monocytogenes, during chilled storage of cottage cheese with fresh- or cultured cream dressing. The mathematical models include the effect of temperature, pH, NaCl, lactic- and sorbic acid and the interaction between these environmental factors. Growth models were developed by combining new and existing cardinal parameter values. Subsequently, the reference growth rate parameters (μref at 25°C) were fitted to a total of 52 growth rates from cottage cheese to improve model performance. The inhibiting effect of mesophilic lactic acid bacteria from added cultures on growth of L. monocytogenes was efficiently modelled using the Jameson approach. The new models appropriately predicted the maximum population density of L. monocytogenes in cottage cheese. The developed models were successfully validated by using 25 growth rates for L. monocytogenes, 17 growth rates for lactic acid bacteria and a total of 26 growth curves for simultaneous growth of L. monocytogenes and lactic acid bacteria in cottage cheese. These data were used in combination with bias- and accuracy factors and with the concept of acceptable simulation zone. Evaluation of predicted growth rates of L. monocytogenes in cottage cheese with fresh- or cultured cream dressing resulted in bias-factors (Bf) of 1.07-1.10 with corresponding accuracy factor (Af) values of 1.11 to 1.22. Lactic acid bacteria from added starter culture were on average predicted to grow 16% faster than observed (Bf of 1.16 and Af of 1.32) and growth of the diacetyl producing aroma culture was on average predicted 9% slower than observed (Bf of 0.91 and Af of 1.17). The acceptable simulation zone method showed the new models to successfully predict maximum population density of L. monocytogenes when growing together with lactic acid bacteria in cottage cheese. 11 of 13 simulations of L

  3. Comparison of human dermal fibroblasts (HDFs) growth rate in culture media supplemented with or without basic fibroblast growth factor (bFGF). (United States)

    Abdian, Narges; Ghasemi-Dehkordi, Payam; Hashemzadeh-Chaleshtori, Morteza; Ganji-Arjenaki, Mahbobe; Doosti, Abbas; Amiri, Beheshteh


    Basic fibroblast growth factor (bFGF or FGF-2) is a member of the FGF family secreted by different kinds of cells like HDFs and it is an important nutritional factor for cell growth and differentiation. The HDFs release bFGF in culture media at very low. The present study aims to investigate the HDFs growth rate in culture media supplemented either with or without bFGF. In brief, HDFs were isolated from human foreskin sample and were cultured in vitro in media containing bFGF and lack of this factor. The cells growth rate was calculated by trypan blue. The karyotyping was performed using G-banding to investigate the chromosomal abnormality of HDFs in both groups. Total RNA of each groups were extracted and cDNA samples were synthesized then, real-time Q-PCR was used to measure the expression level of p27kip1 and cyclin D1 genes normalized to internal control gene (GAPDH). The karyotype analysis showed that HDFs cultured in media or without bFGF had normal karyotype (46 chromosomes, XY) and chromosomal abnormalities were not observed. The cell growth rates in both groups were normal with proliferated exponentially but the slope of growth curve in HDFs cultured in media containing bFGF was increased. Karyotyp test showed that bFGF does not affect on cytogenetic stability of cells. The survey of p27kip1 and cyclin D1 genes by real-time Q-PCR showed that the expression level of these genes were up-regulated when adding bFGF in culture media (p media with growth factor like bFGF could enhance the proliferation and differentiation capacity of cells and improve cells growth rate. Similarly, fibroblast growth factors did not induce any chromosomal abnormality in cells. Furthermore, in HDFs cultured in bFGF supplemented media, the p27kip1 and cyclin D1 genes were up-regulated and suggesting an important role for bFGF in cell-cycle regulation and progression and fibroblast division stimulation. It also suggests that the effects of bFGF on different cell types with

  4. Growth-promoting effect on iron-sulfur proteins on axenic cultures of Entamoeba dispar

    Directory of Open Access Journals (Sweden)

    Khalifa S.A.M.


    Full Text Available A growth-promoting factor (GPF that promotes the growth of Entamoeba dispar under axenic culture conditions was found in fractions of mitochondria (Mt, hydrogenosomes (Hg and chloroplasts (Cp obtained from cells of six different protozoan, mammalian and plant species. We were able to extract the GPF from the Cp-rich leaf cells of a plant (spiderwort: Commelina communis L. in an acetone-soluble fraction as a complex of chlorophyll with low molecular weight proteins (molecular weight [MW] approximately 4,600. We also found that on treatment with 0.6 % complexes of 2-mercapthoethanol (2ME, complexes of chlorophyll-a with iron-sulphur (Fe-S proteins (e.g., ferredoxins [Fd] from spinach and Clostridium pasteurianum and noncomplex rubredoxin (Rd from C. pasteurianum have a growth-promoting effect on E. dispar. These findings suggest that E. dispar may lack a sufficient quantity of some essential components of Fe-S proteins, such as Fe-S center.

  5. [Characteristic of Particulate Emissions from Concrete Batching in Beijing]. (United States)

    Xue, Yi-feng; Zhou, Zhen; Zhong, Lian-hong; Yan, Jing; Qu, Song; Huang, Yu-hu; Tian, He- zhong; Pan, Tao


    With the economic development and population growth in Beijing, there is a strong need for construction and housing, which leads to the increase of the construction areas. Meanwhile, as a local provided material, the production of concrete has been raised. In the process of concrete production by concrete batching, there are numerous particulates emitted, which have large effect on the atmospheric environment, however, systematic study about the tempo-spatial characteristics of pollutant emission from concrete batching is still rare. In this study, we estimated the emission of particulates from concrete batching from 1991 to 2012 using emission factor method, analyzed the tempo-spatial characteristics of pollutant emission, established the uncertainty range by adopting Monte-Carlo method, and predicted the future emission in 2020 based on the relative environmental and economical policies. The results showed that: (1) the emissions of particulates from concrete batching showed a trend of "first increase and then decrease", reaching the maximum in 2005, and then decreased due to stricter emission standard and enhanced environmental management. (2) according to spatial distribution, the emission of particulates from concrete batch mainly concentrated in the urban area with more human activities, and the area between the fifth ring and the sixth ring contributed the most. (3) through scenarios analysis, for further reducing the emission from concrete batching in 2020, more stricter standard for green production as well as powerful supervision is needed.

  6. BatchJS: Implementing Batches in JavaScript

    NARCIS (Netherlands)

    Kasemier, D.


    None of our popular programming languages know how to handle distribution well. Yet our programs interact more and more with each other and our data resorts in databases and web services. Batches are a new addition to languages that can finally bring native support for distribution to our favourite

  7. Analysis of Adiabatic Batch Reactor

    Directory of Open Access Journals (Sweden)

    Erald Gjonaj


    Full Text Available A mixture of acetic anhydride is reacted with excess water in an adiabatic batch reactor to form an exothermic reaction. The concentration of acetic anhydride and the temperature inside the adiabatic batch reactor are calculated with an initial temperature of 20°C, an initial temperature of 30°C, and with a cooling jacket maintaining the temperature at a constant of 20°C. The graphs of the three different scenarios show that the highest temperatures will cause the reaction to occur faster.

  8. Release and degradation of Microcystins during the batch culture of Microcystis aeruginosa and grazing with and without microanimals; Microcystis aeruginosa no baiyo katei oyobi bisho dobutsu no sesshoku katei ni okeru microcystin no yoshutsu to bunkai

    Energy Technology Data Exchange (ETDEWEB)

    Park, H.; Ryu, J. [National Institute of Environmental Research, Seoul (Korea, Republic of); Inamori, Y. [National Inst. for Environmental Studies, Tsukuba (Japan); Jin, I. [Kyungbuk National University, Daegu (Korea, Republic of)


    In order to investigate growth of Microcystis aeruginose, a representative toxic blue algae, and dynamics involved in microcystin degradation, the samples collected in Korean fresh water were cultured. Microcystin is an intracellar toxin, and is retained in a cell as long as it is alive. It is eluted out into surrounding water, when the cell which retains it is dead, and cannot be removed by a general purification treatment. It is therefore essential to remove the algae, when it is abnormally grown, to grasp the dynamics involved in transfer of the toxin resulting from cell breakdown, and to develop the adequate treatment method, to secure safe water. A. hemprichi and M. guttula as a protozoan show high ability of predating the M. aeruginose. Of the systems grazed with microanimals, those with A. hemprichi, P. erythrophthalma and M. guttula show higher degradation ability than the others. It is also found that microcystin is eluted out into surrounding water, when M. aeruginose is abnormally grown in a natural water area and self-degraded, and possibly persists for extended periods. 17 refs., 7 figs., 2 tabs.

  9. Improved growth media and culture techniques for genetic analysis and assessment of biomass utilization by Caldicellulosiruptor bescii. (United States)

    Farkas, Joel; Chung, Daehwan; Cha, Minseok; Copeland, Jennifer; Grayeski, Philip; Westpheling, Janet


    Methods for efficient growth and manipulation of relatively uncharacterized bacteria facilitate their study and are essential for genetic manipulation. We report new growth media and culture techniques for Caldicellulosiruptor bescii, the most thermophilic cellulolytic bacterium known. A low osmolarity defined growth medium (LOD) was developed that avoids problems associated with precipitates that form in previously reported media allowing the monitoring of culture density by optical density at 680 nm (OD(680)) and more efficient DNA transformation by electroporation. This is a defined minimal medium and does not support growth when a carbon source is omitted, making it suitable for selection of nutritional markers as well as the study of biomass utilization by C. bescii. A low osmolarity complex growth medium (LOC) was developed that dramatically improves growth and culture viability during storage, making it a better medium for routine growth and passaging of C. bescii. Both media contain significantly lower solute concentration than previously published media, allowing for flexibility in developing more specialized media types while avoiding the issues of growth inhibition and cell lysis due to osmotic stress. Plating on LOD medium solidified by agar results in ~1,000-fold greater plating efficiency than previously reported and allows the isolation of discrete colonies. These new media represent a significant advance for both genetic manipulation and the study of biomass utilization in C. bescii, and may be applied broadly across the Caldicellulosiruptor genus.

  10. Influence of rat substrain and growth conditions on the characteristics of primary cultures of adult rat spinal cord astrocytes. (United States)

    Codeluppi, Simone; Gregory, Ebba Norsted; Kjell, Jacob; Wigerblad, Gustaf; Olson, Lars; Svensson, Camilla I


    Primary astrocyte cell cultures have become a valuable tool for studies of signaling pathways that regulate astrocyte physiology, reactivity, and function; however, differences in culture preparation affect data reproducibility. The aim of this work was to define optimal conditions for obtaining primary astrocytes from adult rat spinal cord with an expression profile most similar to adult human spinal cord astrocytes. Hence, we examined whether different Sprague-Dawley substrains and culture conditions affect astrocyte culture quality. Medium supplemented with fetal bovine serum from three sources (Sigma, Gibco, Hyclone) or a medium with defined composition (AM medium) was used to culture astrocytes isolated from spinal cords of adult Harlan and Charles River Spraque-Dawley rats. Purity was significantly different between cultures established in media with different sera. No microglia were detected in AM or Hyclone cultures. Gene expression was also affected, with AM cultures expressing the highest level of glutamine synthetase, connexin-43, and glutamate transporter-1. Interestingly, cell response to starvation was substrain dependent. Charles River-derived cultures responded the least, while astrocytes derived from Harlan rats showed a greater decrease in Gfap and glutamine synthetase, suggesting a more quiescent phenotype. Human and Harlan astrocytes cultured in AM media responded similarly to starvation. Taken together, this study shows that rat substrain and growth medium composition affect purity, expression profile and response to starvation of primary astrocytes suggesting that cultures of Harlan rats in AM media have optimal astrocyte characteristics, purity, and similarity to human astrocytes.

  11. [Effects of Ulva pertusa and Gracilaria lemaneiformis on growth of Heterosigma akashiwo (Raphidophyceae) in co-culture]. (United States)

    Wang, You; Yu, Zhi-ming; Song, Xiu-xian; Zhang, Shan-dong


    We studied the effects of fresh tissue and culture medium filtrate of two species of macroalgae, Ulva pertusa (Chlorophyta) and Gracilaria lemaneiformis (Rhodophyta) on growth of Heterosigma akashiwo (Raphidophyceae) in co-culture. Both U. pertusa and G. lemaneiformis, and especially their fresh tissues, significantly impede the growth of H. akashiwo. Carbonate limitations and the presence of environment bacteria are not necessary for the negative effects of macroalgal on H. akashiwo. The simultaneous nutrient assays show that nitrate and phosphate are almost exhausted in the G. lemaneiformis co-culture system, but remain at acceptable levels in the U. pertusa system, when all cells of H. akashiwo are completely dead. When f/2 medium is supplied daily to G. lemaneiformis culture, the growth of H. akashiwo is greatly inhibited but not completely terminated. Furthermore, different amounts of fresh seaweed tissue, and culture medium filtrate prepared from different macroalgal concentrations are analyzed to determine their effects on the growth of H. akashiwo. The results show a positive correlation between the initial macroalgal concentration and the negative effects they exert on the co-cultured microalgae. Results suggest that the allelopathic effects of U. pertusa may be essential for negative effects on H. akashiwo; however, the combined roles of allelopathy and nutrient competition may be responsible for the negative effect of G. lemaneiformis the release of allelochemicals by U. pertusa.

  12. Time dependent physiological characterization of yeast oxidative stress response and growth modulation of protein kinase/phosphatase mutants

    DEFF Research Database (Denmark)

    Altintas, Ali; Workman, Christopher


    The objective of the project was to investigate the time-dependent batch growth effects of oxidative environmental conditions on protein ki nase (PK) and phosphatase (PP) deletion mutants and relevant wild type strains of Saccharomyces cerevisiae . To achieve this goal, 44 different PK and PP.......25, 0.50 and 1.0 mM). Understanding the growth physiology of S. cerevisiae allows us to estimate the link between genotype and stress-response phenotype. Growth physiology parameters, such as growth rate, diauxic shift times, stress-induced stasis times, were measured in fermentative batch cultures...

  13. Cross-cultural correlations of childhood growth and adult breast cancer. (United States)

    Micozzi, M S


    International differences in breast cancer incidence and mortality, and studies on Japanese migrants to the United States, point to the importance of environmental factors, including diet and nutrition, in the etiology of breast cancer. Some studies have suggested that dietary patterns in early life are important to the long-term risk of breast cancer. Given that human growth is partially a function of early dietary intake, cross-cultural correlations between breast cancer rates and anthropometric variables measured at different times in childhood provide additional information about the association of early nutrition and cancer. In this study, the associations between food consumption and anthropometric variables, and childhood growth patterns (attained size at age) and adult breast cancer rates, were considered. Data from cross-sectional growth studies conducted during the years 1956-1971 on children aged 6-18 years were obtained for age-specific stature, sitting height, weight, triceps skinfold thickness, arm and chest circumferences, and biacromial and biiliac diameters. National food consumption data were obtained from the United Nations Food and Agriculture Organization (FAO) and socioeconomic status indicators from the United Nations Children's Fund (UNICEF). Cancer incidence data for the years 1972-1977 were obtained from regional cancer registries reported by the International Agency for Research on Cancer (IARC), and mortality data for 1978 were obtained from national cancer registries around the world. Significant correlations were seen between national food consumption data and childhood growth (attained size at age); between cancer incidence and age-specific stature (r = 0.68), weight (r = 0.59), triceps skinfold thickness (r = 0.78), and biacromial width (r = 0.84); and between mortality and age-specific stature (r = 0.77), weight (r = 0.75), and biacromial width (r = 0.78). In general, the correlation coefficients of the observed anthropometric

  14. Studies on Saccharomyces cerevisiae carrying the plasmid pCYG4 related with ammonia assimilation. Batch experiments. (United States)

    Lima Filho, J L; Ledingham, W M


    Batch culture experiments of three different strains of Saccharomyces cerevisiae have been carried out. The first strain was transformed by a plasmid pCYG4, which carries the glutamate dehydrogenase (NADP-GDH, E.C. 1.4.14) gene conferring an 11-fold increase in activity. The second was transformed by the same plasmid, but without NADP-GDH, and the third was the wild type. The specific growth rates of the two recombinant DNA strains were below that of the wild type, which can be related to extra plasmid protein production.

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

    Directory of Open Access Journals (Sweden)

    Manju Pathak


    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

  16. The role of specific retinoid receptors in sebocyte growth and differentiation in culture. (United States)

    Kim, M J; Ciletti, N; Michel, S; Reichert, U; Rosenfield, R L


    Retinoic acid derivatives (retinoids) exert their pleiotropic effects on cell development through specific nuclear receptors, the retinoic acid receptors and retinoid X receptors. Despite recent progress in understanding the cellular and molecular mechanisms of retinoid activity, it is unknown which of the retinoid receptor pathways are involved in the specific processes of sebocyte growth and development. In this study, we investigated the roles of specific retinoid receptors in sebocyte growth and differentiation, by testing the effects of selective retinoic acid receptor and retinoid X receptor ligands at concentrations between 10-10 M and 10-6 M in a primary rat preputial cell monolayer culture system. Cell growth was determined by number of cells and colonies, and cell differentiation by analysis of lipid-forming colonies. All-trans retinoic acid and selective retinoic acid receptor agonists (CD271 = adapalene, an RAR-beta,gamma agonist; CD2043 = retinoic acid receptor pan-agonist; and CD336 = Am580, an RAR-alpha agonist) caused significant decreases in numbers of cells, colonies, and lipid-forming colonies, but with an exception at high doses of all-trans retinoic acid (10-6 M), with which only a small number of colonies grew but they became twice as differentiated as controls (42.2 +/- 4.0% vs 22.6 +/- 2.7%, mean +/- SEM, lipid-forming colonies, p < 0.01). Furthermore, the RAR-beta,gamma antagonist CD2665 antagonized the suppressive effects of all-trans retinoic acid, adapalene, and CD2043 on both cell growth and differentiation. In contrast, the retinoid X receptor agonist CD2809 increased cell growth slightly and lipid-forming colonies dramatically in a clear dose-related manner to a maximum of 73.7% +/- 6.7% at 10-6 M (p < 0. 001). Our data suggest that retinoic acid receptors and retinoid X receptors differ in their roles in sebocyte growth and differentiation: (i) retinoic acid receptors, especially the beta and/or gamma subtypes, mediate both the

  17. Growth inhibition of cultured marine phytoplankton by toxic algal-derived polyunsaturated aldehydes. (United States)

    Ribalet, François; Berges, John A; Ianora, Adrianna; Casotti, Raffaella


    Several marine diatoms produce polyunsaturated aldehydes (PUAs) that have been shown to be toxic to a wide variety of model organisms, from bacteria to invertebrates. However, very little information is available on their effect on phytoplankton. Here, we expand previous studies to six species of marine phytoplankton, belonging to different taxonomic groups that are well represented in marine plankton. The effect of three PUAs, 2E,4E-decadienal, 2E,4E-octadienal and 2E,4E-heptadienal, was assessed on growth, cell membrane permeability, flow cytometric properties and morphology. A concentration-dependent reduction in the growth rate was observed for all cultures exposed to PUAs with longer-chained aldehydes having stronger effects on growth than shorter-chained aldehydes. Clear differences were observed among the different species. The prymnesiophyte Isochrysis galbana was the most sensitive species to PUA exposure with a lower threshold for an observed effect triggered by mean concentrations of 0.10 micromol L(-1) for 2E,4E-decadienal, 1.86 micromol L(-1) for 2E,4E-octadienal and 3.06 micromol L(-1) for 2E,4E-heptadienal, and a 50% growth inhibition (EC(50)) with respect to the control at 0.99, 2.25 and 5.90 micromol L(-1) for the three PUAs, respectively. Alternatively, the chlorophyte Tetraselmis suecica and the diatom Skeletonema marinoi (formerly S. costatum) were the most resistant species with 50% growth inhibition occurring at concentrations at least two to three times higher than I. galbana. In all species, the three PUAs caused changes in flow cytometric measures of cell size and cell granulosity and increased membrane permeability, assessed using the viability stain SYTOX Green. For example, after 48 h 51.6+/-2.6% of I. galbana cells and 15.0+/-1.8% of S. marinoi cells were not viable. Chromatin fragmentation was observed in the dinoflagellate Amphidinium carterae while clear DNA degradation was observed in the chlorophyte Dunaliella tertiolecta

  18. Regulation of water quality and growth characteristics of indoor raceway culture of Litopenaeus vannamei

    Institute of Scientific and Technical Information of China (English)

    ZANG Weiling; YANG Ming; DAI Xilin; HOU Wenjie; LIU Yongshi; DING Fujiang


    growth data of shrimps throughout the culture period.

  19. Physicochemical Characteristics of Transferon™ Batches. (United States)

    Medina-Rivero, Emilio; Vallejo-Castillo, Luis; Vázquez-Leyva, Said; Pérez-Sánchez, Gilberto; Favari, Liliana; Velasco-Velázquez, Marco; Estrada-Parra, Sergio; Pavón, Lenin; Pérez-Tapia, Sonia Mayra


    Transferon, a biotherapeutic agent that has been used for the past 2 decades for diseases with an inflammatory component, has been approved by regulatory authorities in Mexico (COFEPRIS) for the treatment of patients with herpes infection. The active pharmaceutical ingredient (API) of Transferon is based on polydispersion of peptides that have been extracted from lysed human leukocytes by a dialysis process and a subsequent ultrafiltration step to select molecules below 10 kDa. To physicochemically characterize the drug product, we developed chromatographic methods and an SDS-PAGE approach to analyze the composition and the overall variability of Transferon. Reversed-phase chromatographic profiles of peptide populations demonstrated batch-to-batch consistency from 10 representative batches that harbored 4 primary peaks with a relative standard deviation (RSD) of less than 7%. Aminogram profiles exhibited 17 proteinogenic amino acids and showed that glycine was the most abundant amino acid, with a relative content of approximately 18%. Further, based on their electrophoretic migration, the peptide populations exhibited a molecular mass of about 10 kDa. Finally, we determined the Transferon fingerprint using a mass spectrometry tool. Because each batch was produced from independent pooled buffy coat samples from healthy donors, supplied by a local blood bank, our results support the consistency of the production of Transferon and reveal its peptide identity with regard to its physicochemical attributes.

  20. NDA BATCH 2002-02

    Energy Technology Data Exchange (ETDEWEB)

    Lawrence Livermore National Laboratory


    QC sample results (daily background checks, 20-gram and 100-gram SGS drum checks) were within acceptable criteria established by WIPP's Quality Assurance Objectives for TRU Waste Characterization. Replicate runs were performed on 5 drums with IDs LL85101099TRU, LL85801147TRU, LL85801109TRU, LL85300999TRU and LL85500979TRU. All replicate measurement results are identical at the 95% confidence level as established by WIPP criteria. Note that the batch covered 5 weeks of SGS measurements from 23-Jan-2002 through 22-Feb-2002. Data packet for SGS Batch 2002-02 generated using gamma spectroscopy with the Pu Facility SGS unit is technically reasonable. All QC samples are in compliance with established control limits. The batch data packet has been reviewed for correctness, completeness, consistency and compliance with WIPP's Quality Assurance Objectives and determined to be acceptable. An Expert Review was performed on the data packet between 28-Feb-02 and 09-Jul-02 to check for potential U-235, Np-237 and Am-241 interferences and address drum cases where specific scan segments showed Se gamma ray transmissions for the 136-keV gamma to be below 0.1 %. Two drums in the batch showed Pu-238 at a relative mass ratio more than 2% of all the Pu isotopes.

  1. Physicochemical Characteristics of Transferon™ Batches

    Directory of Open Access Journals (Sweden)

    Emilio Medina-Rivero


    Full Text Available Transferon, a biotherapeutic agent that has been used for the past 2 decades for diseases with an inflammatory component, has been approved by regulatory authorities in Mexico (COFEPRIS for the treatment of patients with herpes infection. The active pharmaceutical ingredient (API of Transferon is based on polydispersion of peptides that have been extracted from lysed human leukocytes by a dialysis process and a subsequent ultrafiltration step to select molecules below 10 kDa. To physicochemically characterize the drug product, we developed chromatographic methods and an SDS-PAGE approach to analyze the composition and the overall variability of Transferon. Reversed-phase chromatographic profiles of peptide populations demonstrated batch-to-batch consistency from 10 representative batches that harbored 4 primary peaks with a relative standard deviation (RSD of less than 7%. Aminogram profiles exhibited 17 proteinogenic amino acids and showed that glycine was the most abundant amino acid, with a relative content of approximately 18%. Further, based on their electrophoretic migration, the peptide populations exhibited a molecular mass of about 10 kDa. Finally, we determined the Transferon fingerprint using a mass spectrometry tool. Because each batch was produced from independent pooled buffy coat samples from healthy donors, supplied by a local blood bank, our results support the consistency of the production of Transferon and reveal its peptide identity with regard to its physicochemical attributes.

  2. Physicochemical Characteristics of Transferon™ Batches (United States)

    Pérez-Sánchez, Gilberto; Favari, Liliana; Estrada-Parra, Sergio


    Transferon, a biotherapeutic agent that has been used for the past 2 decades for diseases with an inflammatory component, has been approved by regulatory authorities in Mexico (COFEPRIS) for the treatment of patients with herpes infection. The active pharmaceutical ingredient (API) of Transferon is based on polydispersion of peptides that have been extracted from lysed human leukocytes by a dialysis process and a subsequent ultrafiltration step to select molecules below 10 kDa. To physicochemically characterize the drug product, we developed chromatographic methods and an SDS-PAGE approach to analyze the composition and the overall variability of Transferon. Reversed-phase chromatographic profiles of peptide populations demonstrated batch-to-batch consistency from 10 representative batches that harbored 4 primary peaks with a relative standard deviation (RSD) of less than 7%. Aminogram profiles exhibited 17 proteinogenic amino acids and showed that glycine was the most abundant amino acid, with a relative content of approximately 18%. Further, based on their electrophoretic migration, the peptide populations exhibited a molecular mass of about 10 kDa. Finally, we determined the Transferon fingerprint using a mass spectrometry tool. Because each batch was produced from independent pooled buffy coat samples from healthy donors, supplied by a local blood bank, our results support the consistency of the production of Transferon and reveal its peptide identity with regard to its physicochemical attributes. PMID:27525277

  3. Batching System for Superior Service (United States)


    Veridian's Portable Batch System (PBS) was the recipient of the 1997 NASA Space Act Award for outstanding software. A batch system is a set of processes for managing queues and jobs. Without a batch system, it is difficult to manage the workload of a computer system. By bundling the enterprise's computing resources, the PBS technology offers users a single coherent interface, resulting in efficient management of the batch services. Users choose which information to package into "containers" for system-wide use. PBS also provides detailed system usage data, a procedure not easily executed without this software. PBS operates on networked, multi-platform UNIX environments. Veridian's new version, PBS Pro,TM has additional features and enhancements, including support for additional operating systems. Veridian distributes the original version of PBS as Open Source software via the PBS website. Customers can register and download the software at no cost. PBS Pro is also available via the web and offers additional features such as increased stability, reliability, and fault tolerance.A company using PBS can expect a significant increase in the effective management of its computing resources. Tangible benefits include increased utilization of costly resources and enhanced understanding of computational requirements and user needs.

  4. Relation of spontaneous transformation in cell culture to adaptive growth and clonal heterogeneity. (United States)

    Rubin, A L; Yao, A; Rubin, H


    Cell transformation in culture is marked by the appearance of morphologically altered cells that continue to multiply to form discrete foci in confluent sheets when the surrounding cells are inhibited. These foci occur spontaneously in early-passage NIH 3T3 cells grown to confluency in 10% calf serum (CS) but are not seen in cultures grown to confluency in 2% CS. However, repeated passage of the cells at low density in 2% CS gives rise to an adapted population that grows to increasingly higher saturation densities and produces large numbers of foci in 2% CS. The increased saturation density of the adapted population in 2% CS is retained upon repeated passage in 10% CS, but the number and size of the foci produced in 2% CS gradually decrease under this regime. Clonal analysis confirms that the focus-forming potential of most if not all of the cells in a population increases in response to a continuously applied growth constraint, although only a small fraction of the population may actually form foci in a given assay. The acquired capacity for focus formation varies widely in clones derived from the adapted population and changes in diverse ways upon further passage of the clones. We propose that the adaptive changes result from progressive selection of successive phenotypic variations in growth capacity that occur spontaneously. The process designated progressive state selection resolves the apparent dichotomy between spontaneous mutation with selection on the one hand and induction on the other, by introducing selection among fluctuating states or metabolic patterns rather than among genetically altered cells.

  5. Low Doses of Celecoxib Stimulate Human Endometrium Growth in A Three-Dimensional Culture Model

    Directory of Open Access Journals (Sweden)

    Mohammad Rasool Khazaei


    Full Text Available Background: The endometrium plays a pivotal role in implantation and pregnancy. CyclooxygenaseII (COX-2 has an important function in biological processes such as cellproliferation and inflammation. Celecoxib is a selective inhibitor of COX-2 with numerouspharmacologic functions. The aim of present study is to investigate the effects ofcelecoxib on the human endometrium in a three-dimensional (3D culture model.Materials and Methods: In this experimental study, normal human endometria (n=10obtained from reproductive age women were cut into 1×1 mm sections. Endometrialexplants were placed between two layers of fibrin gel. To create the fibrin gel, we poureda thin layer of fibrinogen solution [3 mg/ml in medium 199 (M199] into each well of a24-well culture dish and added thrombin enzyme. Endometrial fragments were placed inthe center of each well and covered with a second layer of fibrinogen solution. M199 supplementedwith L-glutamine, fetal bovine serum (FBS, 5% and antibiotics were addedto each well. The media in each experimental well contained either1, 10 or 50 μM ofcelecoxib. At the end of the study, we calculated endometrial tissue growth changes byscoring methods and determined the percentage of angiogenesis. Data were analyzed bythe Kruskal-Wallis method. P<0.05 was considered significant.Results: The growth scores were as follows: control (1.37 ± 0.16, 1 μM (1.96 ± 0.28,10 μM (2.01 ± 0.25, and 50 μM (1.17 ± 0.14 celecoxib, all of which were significantlydifferent. The angiogenesis percentages were: 25.56 ± 6.72% (control, 31.98 ± 6.18% (1μM, 42.67 ± 7.27% (10 μM and 23.44 ± 4.03% (50 μM, which were not significantlydifferent from each other.

  6. Growth inhibition of phytoplankton populations cultured in disphotic zone water by insufficient amounts of dissolved organic carbon

    Energy Technology Data Exchange (ETDEWEB)

    Toyota, T. [Japan Marine Science and Technology Center, Kanagawa (Japan)


    Phytoplankton cultures in disphotic zone water were performed, to examine dissolved organic carbon (DOC) as a controlling agent of the initial lag period and growth rate. Culture media with various concentrations of DOC were prepared by mixing low DOC disphotic zone water with high DOC surface water. Natural phytoplankton populations exhibited strong correlations to the lag period with DOC concentrations and to the population growth rate. Similar tendencies were also confirmed as to a marine diatom dominating in the disphotic zone by culture experiments. By reducing DOC concentrations in seawater samples through pretreatments with ultraviolet radiation, charcoal adsorption and Amberlite resin adsorption, lag periods of diatom increased. Consequently, it was found that the lag period is prolonged in low DOC water. It was suggested that the essential substance to shorten lag periods of phytoplankton cultured in disphotic zone water is a portion of dissolved organic matter. 35 refs., 6 figs., 5 tabs.

  7. The cultural divide: exponential growth in classical 2D and metabolic equilibrium in 3D environments.

    Directory of Open Access Journals (Sweden)

    Krzysztof Wrzesinski

    Full Text Available INTRODUCTION: Cellular metabolism can be considered to have two extremes: one is characterized by exponential growth (in 2D cultures and the other by a dynamic equilibrium (in 3D cultures. We have analyzed the proteome and cellular architecture at these two extremes and found that they are dramatically different. RESULTS: Structurally, actin organization is changed, microtubules are increased and keratins 8 and 18 decreased. Metabolically, glycolysis, fatty acid metabolism and the pentose phosphate shunt are increased while TCA cycle and oxidative phosphorylation is unchanged. Enzymes involved in cholesterol and urea synthesis are increased consistent with the attainment of cholesterol and urea production rates seen in vivo. DNA repair enzymes are increased even though cells are predominantly in Go. Transport around the cell--along the microtubules, through the nuclear pore and in various types of vesicles has been prioritized. There are numerous coherent changes in transcription, splicing, translation, protein folding and degradation. The amount of individual proteins within complexes is shown to be highly coordinated. Typically subunits which initiate a particular function are present in increased amounts compared to other subunits of the same complex. SUMMARY: We have previously demonstrated that cells at dynamic equilibrium can match the physiological performance of cells in tissues in vivo. Here we describe the multitude of protein changes necessary to achieve this performance.


    Directory of Open Access Journals (Sweden)



    Full Text Available The possibility of physiological age and growth regulators affecting callus browning ofcoconut endosperm was investigated. Solid endosperm explants of four coconut fruits fromsame brunches of two coconut cultivars “Samoan Dwarf ” were grown on modified Murashigeand Skoog (MS formula with addition of 10 mg l putresine, 2.50 g l activated charcoal (AC,1.70 g l phytagel, 0, 10 , 10 , 10 , 10 M 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram combined with 10 M 6-benzylaminopurine (BA.Callogenesis occurred on 98.83% of explants. Callus browning between different physiologicalages (antipodal and micropylar tissues of coconut endosperm at 9, 26 and 31 weeks of culture(WOC was significantly different, but not at 16 and 21 WOC. Auxins of 2,4-D and Picloramdid not affect significantly callus browning of endosperm cultures. Auxin doses at 10 , 10 , and10 M decreased significantly callus browning at 9 and 16 WOC, respectively, but at 10 Mbrowning was less significant compared to other doses at 21 WOC. Auxin dose at 10 M causedless significant browning compared to other doses at 31 WOC. The addition of BA decreasedsignificantly callus browning at 9 WOC, but did not affect callus browning thereafter.

  9. Nerve growth factor modulate proliferation of cultured rabbit corneal endothelial cells and epithelial cells. (United States)

    Li, Xinyu; Li, Zhongguo; Qiu, Liangxiu; Zhao, Changsong; Hu, Zhulin


    In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF. MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570 nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner. 50 U/mL and 500 U/mL NGF got more increase of proliferation than that of 5 U/mL NGF did. Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different.

  10. Erythrocytic malaria growth or invasion inhibition assays with emphasis on suspension culture GIA. (United States)

    Haynes, J David; Moch, J Kathleen; Smoot, Douglas S


    Erythrocytic cycle malaria parasite growth or invasion inhibition assays (GIA) compare the effects of various test and control substances on malaria parasite growth in erythrocytes or invasion into erythrocytes in vitro. Although inhibitions by antimalarial drugs in vitro correlate well with drug protective levels required in vivo, as yet there are too few data to know how well inhibitions by antibodies in vitro correlate with the types and degrees of immune protection in vivo. Antibody-mediated GIA is frequently complicated by parasite strain-specific inhibitions, as well as nonspecific inhibitory factors generated in sera collected or stored under nonoptimal conditions. In this chapter, we describe methods for collecting and processing sera, for using different strains of parasite, and a simplified method for staining parasite DNA with Hoechst dye 33342 before quantitating parasites using ultraviolet (UV)-excited flow cytometry. We also describe a new type of GIA using suspension cultures in a 48-well plate. Critical to this method is enclosing the plate in a gassed, heat-sealed plastic bag, which, being low mass, can easily be rested at a 13.5 degrees angle on a rotor platform (114 rpm with 1-in. displacement) to produce gentle pulsatile waves of media in each well. The suspension GIA, which, relative to the static GIA, increased inhibition by one antibody and decreased inhibition by another (Table 1), may better simulate in vivo blood flow and may thus better predict in vivo efficacy.

  11. Effect of Lanthanum on Rice Growth and Physiological Parameters with Split-Root Nutrient Solution Culture

    Institute of Scientific and Technical Information of China (English)

    谢祖彬; 朱建国; 褚海燕; 张雅丽; 高人; 曾青; 曹志洪


    Split-root solution culture was used to study the promoting effect of lanthanum on rice (Oryza sativa) growth and its physiological mechanisms. Results show that low concentration (0.05~1.5 mg*L-1) increases rice yield and grain numbers. High concentration depresses grain formation (9~30 mg*L-1) and root elongation (1.5~30 mg*L-1). No significant influence on straw dry weight was found over the whole concentration range except the 0.05 mg*L-1 treatment. With the increase of La concentration from 0.05 to 0.75 mg*L-1, catalase (CAT) activity in the first fully expandeing leaves and roots decreases. When La concentration is greater than 0.75 mg*L-1 or less than 9 mg*L-1, it significantly decreases superoxide dismutase activity (SOD) in the leaves and roots. No significant effects were found on chlorophyll, protein and malondialdehyde (MDA) content. Possible mechanisms of La′s promoting effect on rice growth and reduction effect of *O-2 were discussed.

  12. Winter Growth of Carps under Different Semi-Intensive Culture Conditions

    Directory of Open Access Journals (Sweden)

    Nadia Nazish* and Abdul Mateen


    Full Text Available The experiment was planned to observe the influence of different semi intensive culture conditions i.e. organic and inorganic fertilizer with rice polish on the growth of carps during winter season. Two earthen ponds were selected and each pond was stocked with Silver carp (Hypophthalmichthys molitrix, Rohu (Labeo rohita and Mori (Cirrhinus mrigala at the ratio of 1:2:1 respectively with a total number of 44 fishes. Pond 1 was treated with poultry dropping and urea while pond 2 was treated with poultry dropping, urea and rice polish. The ponds were treated with at the rate of 0.2 g N/100g of wet body weight of fish. Fertilizers were added on weekly basis while rice polish was added daily. Total net fish production of pond 1 and pond 2 was remained 797.3 and 1033.0 kg/ha/year. The pond treated with fertilizer and artificial feed (rice polish showed 3.6% more fish production than the pond treated only with fertilizer. The physico-chemical parameters were measured on weekly basis. Temperature, light penetration, pH and planktonic biomass showed non-significant difference in both ponds. Pond 2 which was treated with poultry dropping, urea and rice polish showed 1.26 times greater fish growth than pond 1 which was treated with poultry dropping and urea.

  13. Inhibition of the Growth of Plasmodium falciparum in Culture by Stearylamine-Phosphatidylcholine Liposomes

    Directory of Open Access Journals (Sweden)

    Gulam Mustafa Hasan


    Full Text Available We have examined the effect of stearylamine (SA in liposomes on the viability of Plasmodium falciparum in culture by studying the inhibition of incorporation of [3H]-hypoxanthine in the nucleic acid of parasites. Stearylamine in liposomes significantly inhibits the growth of the parasites depending on the phospholipids composition. The maximum inhibition was observed when SA was delivered through Soya phosphatidylcholine (SPC liposomes. The chain length of alkyl group and density of SA in liposomes play a significant role in inhibiting the growth of the parasites. Incorporation of either cholesterol or Distearylphosphatidylethanolamine−Methoxy-Polyethylene glycol-2000 (DSPE-mPEG-2000 in Soya phosphatidylcholine-stearylamine (SPC-SA liposomes improves the efficacy. Intraerythrocytic entry of intact SPC-SA liposomes into infected erythrocytes was visualized using fluorescent microscopy. No hemolysis was observed in uninfected erythrocytes, and slight hemolysis was noted in infected erythrocytes at high concentrations of SPC-SA liposomes. Overall, our data suggested SA in SPC-liposomes might have potential application in malaria chemotherapy.

  14. In vitro development of bovine secondary follicles in two- and three-dimensional culture systems using vascular endothelial growth factor, insulin-like growth factor-1, and growth hormone. (United States)

    Araújo, V R; Gastal, M O; Wischral, A; Figueiredo, J R; Gastal, E L


    The aim of this study was to evaluate the development and estradiol production of isolated bovine secondary follicles in two-dimensional (2D, experiment 1) and three-dimensional (3D using alginate, experiment 2) long-term culture systems in the absence (control group; only α-MEM(+)) or presence of vascular endothelial growth factor (VEGF), insulin-like growth factor-1, or GH alone, or a combination of all. A total of 363 isolated secondary follicles were cultured individually for 32 days at 38.5 °C in 5% CO2 in a humidified incubator with addition of medium (5 μL) every other day. In 2D culture system, follicular growth and antrum formation rates were higher (P 0.05). In summary, this study demonstrated that the benefits of using a certain type of medium supplement depended on the culture system (2D vs. 3D). Vascular endothelial growth factor was an effective supplement for the in vitro culture of bovine secondary follicles when the 2D culture system was used, whereas GH only affected estradiol production using the 3D culture system. This study sheds light on advancements in methodology to facilitate subsequent studies on bovine preantral follicle development.

  15. Bile salts inhibit growth and induce apoptosis of culture human normal esophageal mucosal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Ru Zhang; Jun Gong; Hui Wang; Li Wang


    AIM: To investigate the effect of six bile salts:glycocholate (GC), glycochenodeoxycholate (GCDC),glycodeoxycholate (GDC), taurocholate (TC),taurochenodeoxycholate (TCDC), taurodeoxycholate (TDC), and their mixture on cultured human normal esophageal mucosal epithelial cells.METHODS: Human normal esophageal mucosal epithelial cells were cultured with serum-free keratinocyte medium. 3-[4,5-Dimethylthiaolyl]-2,5-diphenyl-tetrazolium bromide assay was applied to the detection of cell proliferation. Apoptotic morphology was observed by phase-contrast video microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Sub-G1 DNA fragmentations and early apoptotic cells were assayed by flow cytometry (FCM) with propidium iodide (PI) staining and annexin V-FITC conjugated with PI staining.Apoptotic DNA ladders on agarose gel electrophoresis were observed.RESULTS: Except for GC, GCDC, GDC, TC, TCDC, TDC and their mixture could initiate growth inhibition of esophageal mucosal epithelial cells in a dose- and time-dependent manner. TUNEL and FCM assays demonstrated that the bile salts at 500 μmol/L and their mixture at 1 500 μmol/L induced apoptosis except for GC. The percentage of sub-G1 detected by FCM with PI staining was 83.5% in cells treated with 500μmol/L TC for 2 h, and 19.8%, 20.4%, 25.6%, 13.5%, and 75.8% in cells treated with 500 μmol/L GCDC, TCDC, GDC,TDC, and 1 500 μmol/L mixture for 24 h, respectively,which were higher than that of the control (1.5%). The percentage was 1.4% in cells with 500 μmol/L GC for 24 h.DNA ladders on agarose gel electrophoresis were seen in cells treated with 500 μmol/L TC for 2 h and 1 500 μmol/Lmixture for 24 h.CONCLUSION: All GCDC, GDC, TC, TCDC, TDC and their mixture can inhibit growth and induce apoptosis of cultured human normal esophageal mucosal epithelial cells, but GC is well tolerated by the cells.

  16. Changes in auxin level in the course of growth of a sunflower crown-gall suspension culture

    Directory of Open Access Journals (Sweden)

    Zofia Chirek


    Full Text Available The auxin level in the cell mass and culture medium was determined by means of the Avena straight caleoptile test in various periods of the suspension culture cycle of the sunflower crown-gall tumour. The investigations were performed in the course of the zero passage (PO and first one (Pl, differing in their time of duration of maximum growth and its intensity. In both passages the intra- and extra-cellular auxin levels reach values of the same order. At the beginning of the maximal growth phase the activity corresponding to IAA in the cells prevails over that of the other auxin-like compounds. This disproportion diminishes with further development of the culture, and with the beginning of the stationary phase the cellular IAA level is lower than that of the remaining auxin-like compounds. The short phase of maximal growth (PO occurs with an auxin level decreasing in the cell mass and increasing in the medium, and towards the end of the cycle these levels become equal. During the long phase of maximal growth (Pl the total amount of auxins in the cells increases and is 2-3 times higher than in the medium, whereas IAA in the cells remains at a constant level. These results suggest that the participation of IAA in the intracellular pool of auxin-like substances is decisive for the mitotic activity of the cells and maintenance of growth in the culture.

  17. NGBAuth - Next Generation Batch Authentication for long running batch jobs.

    CERN Document Server

    Juto, Zakarias


    This document describes the prototyping of a new solution for the CERN batch authentication of long running jobs. While the job submission requires valid user credentials, these have to be renewed due to long queuing and execution times. Described within is a new system which will guarantee a similar level of security as the old LSFAuth while simplifying the implementation and the overall architecture. The new system is being built on solid, streamlined and tested components (notably OpenSSL) and a priority has been to make it more generic in order to facilitate the evolution of the current system such as for the expected migration from LSF to Condor as backend batch system.

  18. Effects of High Ammonium Concentration on Growth and Nutrient Uptake of Lettuce Plants with Solution Culture

    Institute of Scientific and Technical Information of China (English)


    A nutrition solution experiment was conducted over two months to investigate the response of vegetable crops to high concentrations of ammonium, using lettuce (Lactuca sativa L. cv. Angustana Irish) as a test crop. Ammonium concentrations were designed in 5 levels, ranging from 12 mmol N L-1 to 22 mmol N L-1 and local tap water was used as water source. At the first culture stage (0-9 days), lettuce plants maintained normal growth while the lettuce roots were increasingly impaired. During the subsequent three stages the root structure was greatly damaged, and roots became brown or black through continuous supply of high concentration of ammonium. However, there was no obvious reduction of the aboveground biomass of the plants in the high ammonium treatments compared to those supplied with nitrate alone. In contrast to results obtained in another experiment from us with distilled water, the detrimental effect of high ammonium concentration on lettuce growth was greatly alleviated. Based on the results, it was postulated that the small amount of nitrate and the higher amount of bicarbonate existed in the tap water might mitigate the adverse effects of high ammonium N. The higher bicarbonate content in water and soil has usually been regarded as a major constraint factor limiting plant growth in calcareous soil areas. However, the reaction of bicarbonate to ammonium might produce positively interactive effect on reduction of both damages. The lettuce plants grown in ammonium solutions took up less P, K, Fe, Mn and Cu and more Ca than those grown in the nitrate nutrient solution. In conclusion, the results indicated that the N form imposed an obvious influence on absorption of cations and anions. Supplying ammonium-N stimulated transport of Ca, Mg and Mn to shoots of lettuce.

  19. Batch-related sterile endophthalmitis following intravitreal injection of bevacizumab

    Directory of Open Access Journals (Sweden)

    Morteza Entezari


    Full Text Available Background: To report a series of patients with sterile endophthalmitis after intravitreal bevacizumab (IVB injection from 2 different batches of bevacizumab. Materials and Methods: Records of 11 eyes with severe inflammation after IVB injections from two different batches (7 eyes from one and 4 from the other on two separate days were evaluated. Fifteen eyes of 15 patients in one day were treated with one batch and 18 eyes of 17 patients were treated another day using another batch injected for different retinal diseases. Each batch was opened on the day of injection. We used commercially available bevacizumab (100 mg/4 ml kept at 4°C. Severe cases with hypopyon were admitted to the ward and underwent anterior chamber and vitreous tap for direct smear and culture. Results: Pain, redness and decreased vision began after 11-17 days. All had anterior chamber and vitreous reactions and 5 had hypopyon. Antibiotics and corticosteroids were initiated immediately, but the antibiotics were discontinued after negative culture results. Visual acuity returned to pre-injection levels in 10 eyes after 1 month and only in one eye pars plana vitrectomy was performed. Mean VA at the time of presentation with inflammation (1.76 ± 0.78 logMAR decreased significantly (P = 0.008 compared to the initial mean corrected VA (1.18 ± 0.55 logMAR; however, final mean corrected VA (1.02 ± 0.48 logMAR improved in comparison with the baseline but not to a significant level (P = 0.159. Conclusions: We report a cluster of sterile endophthalmitis following intravitreal injection of bevacizumab from the same batch of bevacizumab that has a favorable prognosis.

  20. Trickle water and feeding system in plant culture and light-dark cycle effects on plant growth (United States)

    Takano, T.; Inada, K.; Takanashi, J.

    Rockwool, as an inert medium covered or bagged with polyethylene film, can be effectively used for plant culture in space station. The most important machine is the pump adjusting the dripping rate in the feeding system. Hydro-aeroponics may be adaptable to a space laboratory. The shortening of the light-dark cycles inhibits plant growth and induces an abnormal morphogenesis. A photoperiod of 12-hr-dark may be needed for plant growth.

  1. Mixed-culture transcriptome analysis reveals the molecular basis of mixed-culture growth in Streptococcus thermophilus and Lactobacillus bulgaricus

    NARCIS (Netherlands)

    Sieuwerts, S.; Molenaar, D.; Hijum, van S.A.F.T.; Beerthuyzen, M.; Stevens, M.J.A.; Janssen, P.W.; Ingham, C.J.; Bok, de F.A.M.; Vos, de W.M.; Hylckama Vlieg, van J.E.T.


    Many food fermentations are performed using mixed cultures of lactic acid bacteria. Interactions between strains are of key importance for the performance of these fermentations. Yogurt fermentation by Streptococcus thermophilus and Lactobacillus bulgaricus (basonym, Lactobacillus delbrueckii subsp.

  2. Versatile modeling and optimization of fed batch processes for the production of secreted heterologous proteins with Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Gasser Brigitte


    Full Text Available Abstract Background Secretion of heterologous proteins depends both on biomass concentration and on the specific product secretion rate, which in turn is not constant at varying specific growth rates. As fed batch processes usually do not maintain a steady state throughout the feed phase, it is not trivial to model and optimize such a process by mathematical means. Results We have developed a model for product accumulation in fed batch based on iterative calculation in Microsoft Excel spreadsheets, and used the Solver software to optimize the time course of the media feed in order to maximize the volumetric productivity. The optimum feed phase consisted of an exponential feed at maximum specific growth rate, followed by a phase with linearly increasing feed rate and consequently steadily decreasing specific growth rate. The latter phase could be modeled also by exact mathematical treatment by the calculus of variations, yielding the explicit shape of the growth function, however, with certain indeterminate parameters. To evaluate the latter, one needs a numerical optimum search algorithm. The explicit shape of the growth function provides additional evidence that the Excel model results in correct data. Experimental evaluation in two independent fed batch cultures resulted in a good correlation to the optimized model data, and a 2.2 fold improvement of the volumetric productivity. Conclusion The advantages of the procedure we describe here are the ease of use and the flexibility, applying software familiar to every scientist and engineer, and rapid calculation which makes predictions extremely easy, so that many options can be tested in silico quickly. Additional options like further biological and technological constraints or different functions for specific productivity and biomass yield can easily be integrated.

  3. Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base CELSS (Controlled Ecological Life Support System) (United States)

    Venketeswaran, S.


    Experiments were carried out on plant tissue cultures, seed germination, seedling development and plants grown on Simulated Lunar Soil to evaluate the potential of future development of lunar based agriculture. The studies done to determine the effect of the placement of SLS on tissue cultures showed no adverse effect of SLS on tissue cultures. Although statistically insignificant, SLS in suspension showed a comparatively higher growth rate. Observations indicate the SLS, itself cannot support calli growth but was able to show a positive effect on growth rate of calli when supplemented with MS salts. This positive effect related to nutritive value of the SLS was found to have improved at high pH levels, than at the recommended low pH levels for standard media. Results from seed germination indicated that there is neither inhibitory, toxicity nor stimulatory effect of SLS, even though SLS contains high amounts of aluminum compounds compared to earth soil. Analysis of seeding development and growth data showed significant reduction in growth rate indicating that, SLS was a poor growth medium for plant life. This was confirmed by the studies done with embryos and direct plant growth on SLS. Further observations attributed this poor quality of SLS is due to it's lack of essential mineral elements needed for plant growth. By changing the pH of the soil, to more basic conditions, the quality of SLS for plant growth could be improved up to a significant level. Also it was found that the quality of SLS could be improved by almost twice, by external supply of major mineral elements, directly to SLS.

  4. Batch-oriented software appliances

    CERN Document Server

    Murri, Riccardo


    This paper presents AppPot, a system for creating Linux software appliances. AppPot can be run as a regular batch or grid job and executed in user space, and requires no special virtualization support in the infrastructure. The main design goal of AppPot is to bring the benefits of a virtualization-based IaaS cloud to existing batch-oriented computing infrastructures. In particular, AppPot addresses the application deployment and configuration on large heterogeneous computing infrastructures: users are enabled to prepare their own customized virtual appliance for providing a safe execution environment for their applications. These appliances can then be executed on virtually any computing infrastructure being in a private or public cloud as well as any batch-controlled computing clusters the user may have access to. We give an overview of AppPot and its features, the technology that makes it possible, and report on experiences running it in production use within the Swiss National Grid infrastructure SMSCG.

  5. NDA Batch 2002-13

    Energy Technology Data Exchange (ETDEWEB)

    Hollister, R


    QC sample results (daily background check drum and 100-gram SGS check drum) were within acceptance criteria established by WIPP's Quality Assurance Objectives for TRU Waste Characterization. Replicate runs were performed on drum LL85501243TRU. Replicate measurement results are identical at the 95% confidence level as established by WIPP criteria. HWM NCAR No. 02-1000168 issued on 17-Oct-2002 regarding a partially dislodged Cd sheet filter on the HPGe coaxial detector. This physical geometry occurred on 01-Oct-2002 and was not corrected until 10-Oct-2002, during which period is inclusive of the present batch run of drums. Per discussions among the Independent Technical Reviewer, Expert Reviewer and the Technical QA Supervisor, as well as in consultation with John Fleissner, Technical Point of Contact from Canberra, the analytical results are technically reliable. All QC standard runs during this period were in control. Data packet for SGS Batch 2002-13 generated using passive gamma-ray spectroscopy with the Pu Facility SGS unit is technically reasonable. All QC samples are in compliance with establiShed control limits. The batch data packet has been reviewed for correctness, completeness, consistency and compliance with WIPP's Quality Assurance Objectives and determined to be acceptable.

  6. Growth performance, fillet quality, and reproductive maturity of rainbow trout (Oncorhynchus mykiss) cultured to 5 kilograms within freshwater recirculating systems (United States)

    Rainbow trout are commonly cultured within aquaculture systems to one pound or less and marketed as pan-sized fillets. Production of larger rainbow trout provides a distinguishable product. Research that describes the growth performance and fillet quality of large rainbow trout is limited, particula...

  7. Effects of Culture Conditions, Carbon Source and Regulators on Saffron Callus Growth and Crocin Accumulation in the Callus

    Institute of Scientific and Technical Information of China (English)

    刘雪; 郭志刚; 刘瑞芝


    There are many factors influencing the growth and secondary metabolites of callus and saffron callus. In this paper, the effects of culture conditions, including culture temperatures, light levels, the carbon source and its concentration, and the preserve of regulators (mainly hormones), are studied for callus cultures. All the experiments used Murashige and Skoog (MS) solid medium as the basic medium with 10 g/L agar, pH 5.75.8. Saffron callus was cultured at 20℃ in the dark, with a sucrose concentration of 45 g/L (or starchy hydrolysate concentration of 40 g/L), but 30 g/L sucrose was best for the synthesis of crocin (for starchy hydrolysate the concentration can range from 20 to 40 g/L). To promote callus growth, the best auxin was α-naphthaleneacetic acid (NAA) and the optimum ratio of NAA (mg/L) to benzylaminopurine (BA) (mg/L) was and uniconazole (S-07) (1.25 mg/L) increased the crocin content remarkably as analyzed by high performance liquid chromatography (HPLC). NAA (2 mg/L) promoted the growth of saffron callus but had no benefit and may inhibit crocin synthesis while S-07 (1.25 mg/L) had the opposite effect. GA3 promoted both growth and synthesis.

  8. Discoordinate regulation of different K channels in cultured rat skeletal muscle by nerve growth factor. (United States)

    Vigdor-Alboim, S; Rothman, C; Braiman, L; Bak, A; Langzam, L; Yosef, O; Sterengarz, B B; Nawrath, H; Brodie, C; Sampson, S R


    We investigated the effects of nerve growth factor (NGF) on expression of K+ channels in cultured skeletal muscle. The channels studied were (1) charybdotoxin (ChTx)-sensitive channels by using a polyclonal antibody raised in rabbits against ChTx, (2) Kv1.5 voltage-sensitive channels, and (3) apamin-sensitive (afterhyperpolarization) channels. Crude homogenates were prepared from cultures made from limb muscles of 1-2-day-old rat pups for identification of ChTx-sensitive and Kv1.5 channels by Western blotting techniques. Apamin-sensitive K+ channels were studied by measurement of specific [125I]-apamin binding by whole cell preparations. ChTx-sensitive channels display a fusion-related increase in expression, and NGF downregulates these channels in both myoblasts and myotubes. Voltage-dependent Kv1.5 channel expression is low in myoblasts and increases dramatically with fusion; NGF induces early expression of these channels and causes expression after fusion to increase even further. NGF downregulates apamin-sensitive channels. NGF increases the rate of fall of the action potential recorded intracellularly from single myotubes with intracellular microelectrodes. The results confirm and extend those of previous studies in showing a functional role for NGF in the regulation of membrane properties of skeletal muscle. Moreover, the findings demonstrate that the different K+ channels in this preparation are regulated in a discoordinate manner. The divergent effects of NGF on expression of different K+ channels, however, do not appear sufficient to explain the NGF-induced increase in the rate of fall of the action potential. The changes during the falling phase may rather be due to increases in channel properties or may result from an increased driving force on the membrane potential secondary to the NGF-induced hyperpolarization.

  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


    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. 间歇结晶过程的分批优化%Batch-to-batch Optimization of Batch Crystallization Processes

    Institute of Scientific and Technical Information of China (English)

    Woranee Paengjuntuek; Paisan Kittisupakorn; Amornchai Arpornwichanop


    It is the fact that several process parameters are either unknown or uncertain. Therefore, an optimal control profile calculated with developed process models with respect to such process parameters may not give an optimal performance when implemented to real processes. This study proposes a batch-to-batch optimization strat-egy for the estimation of uncertain kinetic parameters in a batch crystallization process of potassium sulfate produc-tion. The knowledge of a crystal size distribution of the product at the end of batch operation is used in the proposedmethodology. The updated kinetic parameters are applied for determining an optimal operating temperature policy for the next batch run.

  11. Managing the perfect storm: The role of founders in creating organizational culture in high-technology, high-growth startup companies


    Paajanen, Nils


    Culture is an important factor in the performance of organizations. The role of culture is magnified in high-tech, high growth startup companies due their operating environment as defined by extreme uncertainty. Culture in this context is seen to provide stability ensuring the organizations external ability to survive and ensuring internal cohesion. Organizational culture is also affected by path-dependency. This is why the role of founders in moulding organizational culture at the very outse...

  12. Algal growth and community structure in a mixed-culture system using coal seam gas water as the water source. (United States)

    Buchanan, Jessica J; Slater, Frances R; Bai, Xue; Pratt, Steven


    Coal seam gas (CSG) is being touted as a transition fuel as the world moves towards low-carbon economies. However, the development of CSG reserves will generate enormous volumes of saline water. In this work, we investigate the potential of using this saline water to support mass algae production. Water and brine from a CSG water treatment facility (1.6 and 11.6 g total dissolved solids per litre (TDS L(-1)) respectively) were inoculated with algal biomass from freshwater and seawater environments and supplemented with nutrients in open, fed-batch reactors. Significant algal growth was recorded, with maximum specific growth rates in CSG water and CSG brine of 0.20 +/- 0.05 d(-1) and 0.26 +/- 0.04 d(-1) respectively. These maximum specific growth rates were equal to or greater than specific growth rates in deionized water and seawater diluted to the same salinity. However, algal growth lag time in CSG brine was between 7 and 9 times longer than in other waters. Microscopy and terminal-restriction fragment length polymorphism (T-RFLP) were used to monitor community structure in the reactors. The same few algal species dominated all of the reactors, except for the CSG brine reactor at day 15. This result indicates that conditions in CSG brine select for different species of algae compared to seawater of the same salinity and other waters tested. The findings suggest that mass algae production in CSG water is feasible but algae community composition may be a function of CSG water chemistry. This has implications for the downstream use of algae.

  13. Effect of Bifidobacterium upon Clostridium difficile growth and toxicity when co-cultured in different prebiotic substrates

    Directory of Open Access Journals (Sweden)

    Lorena Valdés Varela


    Full Text Available The intestinal overgrowth of Clostridium difficile, often after disturbance of the gut microbiota by antibiotic treatment, leads to C. difficile infection (CDI which manifestation ranges from mild diarrhoea to life-threatening conditions. The increasing CDI incidence, not only in compromised subjects but also in traditionally considered low-risk populations, together with the frequent relapses of the disease, has attracted the interest for prevention/therapeutic options. Among these, probiotics, prebiotics or synbiotics constitute a promising approach. In this study we determined the potential of selected Bifidobacterium strains for the inhibition of C. difficile growth and toxicity in different carbon sources. We conducted co-cultures of the toxigenic strain C. difficile LMG21717 with four Bifidobacterium strains (Bifidobacterium longum IPLA20022, Bifidobacterium breve IPLA20006, Bifidobacterium bifidum IPLA20015, and Bifidobacterium animalis subsp. lactis Bb12 in the presence of various prebiotic substrates (Inulin, Synergy and Actilight or glucose, and compared the results with those obtained for the corresponding mono-cultures. C. difficile and bifidobacteria levels were quantified by qPCR; the pH and the production of short chain fatty acids was also determined. Moreover, supernatants of the cultures were collected to evaluate their toxicity using a recently developed model. Results showed that co-culture with B. longum IPLA20022 and B. breve IPLA20006 in the presence of short-chain fructooligosaccharides, but not of Inulin, as carbon source significantly reduced the growth of the pathogen. With the sole exception of B. animalis Bb12, whose growth was enhanced, the presence of C. difficile did not show major effects upon the growth of the bifidobacteria. In accordance with the growth data, B. longum and B. breve were the strains showing higher reduction in the toxicity of the co-culture supernatants.

  14. Effects of photoperiod, plant growth regulators and culture media on in vitro growth of seedlings of Cyrtochilum loxense (Lindl. Kraenzl. an endemic and endangered orchid from Ecuador

    Directory of Open Access Journals (Sweden)

    Yadira González


    Full Text Available Cyrtochilum loxense (Lindl. Kraenzl. is an endemic and seriously endangered orchid species endemic in the Loja Province (Southern Ecuador. The main goals of this research were to analyze how culture media, plant growth regulators and photoperiod affect the growth of C. loxense. Eight month old plants (approximate 1 – 1.5 cm in height obtained by in vitro germination, were cultivated on MS media or Knudson C; MS with three levels of naphthalene acetic acid (NAA and 6-benzylaminopurine (BAP (2/0.5; 1/0.5 y 0.5/ 0.5 mg-1L; and three photoperiodic regimes (24/0, 16/8, 8/16 h on MS with and without plant growth regulators. No significant differences of shoot induction were observed on media with or without plant growth regulators, and all tested photoperiods. The highest growth (1.2 cm was observed in plantlets cultivated on growth regulator-free media with a 16/8 photoperiod. Also the shoot and root formation was better in this species in absence of plant growth regulators. Probably this response is due to the endogenous hormone levels in the tissues or due to the kind and concentrations of PGRs used were too low to induce positive morphogenetic responses.


    directed to HeLa cells . The following results were obtained: Of those cultures that were susceptible, three different patterns of dose response were...obtained that were interpreted as different possible manifestations of interference. HeLa cells , which were among the least sensitive to low infection

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

    Directory of Open Access Journals (Sweden)

    Williams Sylvain


    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.

  17. Growth response and survival of Heterobranchus longifilis cultured at different water levels in outdoor concrete tanks

    Directory of Open Access Journals (Sweden)

    Anthony A. Nlewadim


    Full Text Available Thirteen-day-old hatchery-raised fry obtained from hormonally-induced spawns of mature African catfish H. longifilis brood stock were introduced to three different water levels (0.35, 0.50 and 0.65m in four replicates in 12 units of 2x2x1m3 outdoor concrete tanks. The fry were similarly stocked initially at 50 fry m-2 and later thinned down to 5 fish m-2 and cultured for 6 months. Fish were fed twice daily with commercial pellet feeds (Coppens™ while adjusting the feeding rate from 10 to 4% body weight and pellet size from 0.2 to 4.5 mm. The effects of pond water levels were evaluated in growth responses and survival. Water quality variables were similar (p > 0.05 in all compartments. Temperature, dissolved oxygen and pH were at the optimum level for fish. The results reveal significant (P H. longifilis from fry to sub-adult and from the grow-out/fattening of sub-adult to adult, respectively, in outdoor concrete tanks.

  18. Organic and inorganic lead inhibit neurite growth in vertebrate and invertebrate neurons in culture. (United States)

    Audesirk, G; Shugarts, D; Nelson, G; Przekwas, J


    Neurons from brains of chick embryos and pond snails (Lymnaea stagnalis) were cultured for 3 to 4 d in the presence of no toxins, inorganic lead (PbCl2), or organic lead (triethyl lead chloride). In chick neurons, inorganic lead reduced the percentage of cells that grew neurites (IC50 = 270 microM total lead, approximately 70 nM free Pb2+) but did not reduce the number of neurites per cell or the mean neurite length. Triethyl lead reduced the percentage of cells that grew neurites (IC50 = 0.24 microM) and the mean neurite length (extrapolated IC50 = 3.6 microM) but did not reduce the number of neurites per cell. In Lymnaea neurons, inorganic lead reduced the percentage of cells that grew neurites (IC50 = 13 microM total lead; approximately 10 nM free Pb2+). Triethyl lead reduced the percentage of cells that grew neurites (IC50 = 0.4 microM) and exerted significant toxicity at 0.2 microM. The two forms of lead affected neurite growth in qualitatively different ways, which suggests that their mechanisms of action are different.

  19. Induction of vascular endothelial growth factor by nitric oxide in cultured human articular chondrocytes. (United States)

    Turpaev, K; Litvinov, D; Dubovaya, V; Panasyuk, A; Ivanov, D; Prassolov, V


    We investigated the role of nitric oxide (NO) in the control of vascular endothelial growth factor A (VEGF) gene expression in cultured human articular chondrocytes. Cell treatment with the NO-generating compound nitrosoglutathione (GSNO) caused a significant accumulation of 4.4 kb VEGF mRNA, a major VEGF mRNA isoform expressing in chondrocytes. This is the first demonstration that NO can induce VEGF mRNA expression in chondrocytes. VEGF mRNA level was not affected in cells exposed to dibutyryl cGMP, a non-hydrolyzable analog of cGMP, suggesting that the cGMP system is not involved in NO-dependent transcriptional activation of VEGF gene. The GSNO-stimulated induction of VEGF mRNA was slightly attenuated by MAP protein kinase inhibitors PD98058 and SB203580, but was completely blocked in cells incubated with GSNO in the presence of catalase and superoxide dismutase, enzymes scavenging reactive oxygen species (ROS), or in the presence of thiol-containing antioxidants, N-acetyl cysteine and reduced glutathione. These results suggest that in articular chondrocytes the GSNO-induced VEGF gene transcriptional activation is dependent on endogenous ROS production and oxidative thiol modifications.

  20. Impact of salinity and growth phase on alkenone distributions in coastal haptophytes

    NARCIS (Netherlands)

    Chivall, D.; M'Boule, D.; Sinke-Schoen, D.; Sinninghe Damsté, J.S.; Schouten, S.; van der Meer, M.T.J.


    Batch cultures of Isochrysis galbana (strain CCMP 1323) and Chrysotila lamellosa (strain CCMP 1307) were grown at salinity values of ca. 10 to ca. 35 and the alkenone distributions determined for different growth phases. U-37(K ') values decreased slightly with salinity for C. lamellosa but were lar

  1. Growth-expectations among women entrepreneurs: embedded in networks and culture in Algeria, Morocco, Tunisia and in Belgium and France

    DEFF Research Database (Denmark)

    Cheraghi, Maryam; Setti, Zakia; Schøtt, Thomas


    and secular-rational culture differ in roles for women, which influence women entrepreneurs' networking and expectations. The design compares cultures, with data from three traditional societies, Algeria, Morocco and Tunisia and two secular-rational societies, France and Belgium, surveyed in the Global...... Entrepreneurship Monitor, randomly sampling 39,336 women, including 2,306 entrepreneurs. Analyses show that women entrepreneurs have growth-expectations based on their background and increased by their competence and opportunity-motive, which also promote business networks around their firms. Formation......An entrepreneur usually has an expectation for the firm, expecting expansion, stability or contraction. Expectation is influenced by the entrepreneur's attributes, but expectation is also embedded in the micro-environment of networking and the macro-environment of culture. Traditional culture...

  2. Artificial neural network-based model for the prediction of optimal growth and culture conditions for maximum biomass accumulation in multiple shoot cultures of Centella asiatica. (United States)

    Prasad, Archana; Prakash, Om; Mehrotra, Shakti; Khan, Feroz; Mathur, Ajay Kumar; Mathur, Archana


    An artificial neural network (ANN)-based modelling approach is used to determine the synergistic effect of five major components of growth medium (Mg, Cu, Zn, nitrate and sucrose) on improved in vitro biomass yield in multiple shoot cultures of Centella asiatica. The back propagation neural network (BPNN) was employed to predict optimal biomass accumulation in terms of growth index over a defined culture duration of 35 days. The four variable concentrations of five media components, i.e. MgSO4 (0, 0.75, 1.5, 3.0 mM), ZnSO4 (0, 15, 30, 60 μM), CuSO4 (0, 0.05, 0.1, 0.2 μM), NO3 (20, 30, 40, 60 mM) and sucrose (1, 3, 5, 7 %, w/v) were taken as inputs for the ANN model. The designed model was evaluated by performing three different sets of validation experiments that indicated a greater similarity between the target and predicted dataset. The results of the modelling experiment suggested that 1.5 mM Mg, 30 μM Zn, 0.1 μM Cu, 40 mM NO3 and 6 % (w/v) sucrose were the respective optimal concentrations of the tested medium components for achieving maximum growth index of 1654.46 with high centelloside yield (62.37 mg DW/culture) in the cultured multiple shoots. This study can facilitate the generation of higher biomass of uniform, clean, good quality C. asiatica herb that can efficiently be utilized by pharmaceutical industries.

  3. Modeling of olive oil degradation and oleic acid inhibition during chemostat and batch cultivation of Bacillus thermoleovorans IHI-91. (United States)

    Becker, P; Märkl, H


    Olive oil degradation by the thermophilic lipolytic strain Bacillus thermoleovorans IHI-91 in chemostat and batch culture was modeled to obtain a general understanding of the underlying principles and limitations of the process and to quantify its stoichiometry. Chemostat experiments with olive oil as the sole carbon source were successfully described using the Monod chemostat model extended by terms for maintenance requirements and wall growth. Maintenance requirements and biomass yield coefficients were in the range reported for mesophiles. For a chemostat experiment at D = 0.3 h(-1) the model was validated up to an olive oil feed concentration of about 3.0 g L(-1) above which an inhibitory effect occurred. Further analysis showed that the liberated oleic acid is the main cause for this inhibition. Using steady-state oleic acid concentrations measured in chemostat experiments with olive oil as substrate it was possible to derive a kinetic expression for oleic acid utilization, showing that a concentration of 430 mg L(-1) leads to a complete growth inhibition. Oleic acid accumulation observed during batch fermentations can be predicted using a model involving growth-associated lipase production and olive oil hydrolysis. Simulations confirmed that this accumulation is the cause for the sudden growth cessation occurring in batch fermentations with higher olive oil start concentrations. Further, an oscillatory behavior, as observed in some chemostat experiments, can also be predicted using the latter model. This work clearly demonstrates that thermophilic lipid degradation by Bacillus thermoleovorans IHI-91 is limited by long-chain fatty acid beta-oxidation rather than oil hydrolysis.

  4. Growth hormone-releasing factor regulates growth hormone mRNA in primary cultures of rat pituitary cells.


    Gick, G G; Zeytin, F N; BRAZEAU, P.; Ling, N C; Esch, F S; Bancroft, C


    A peptide with high intrinsic activity for specifically stimulating the secretion of immunoreactive growth hormone (GH; somatotropin) has been characterized and reproduced by total synthesis. This peptide, human pancreatic growth hormone-releasing factor, 44-amino-acid form (hpGRF1-44-NH2), was isolated from a tumor localized in the pancreas of a patient with acromegaly. We report here the effect of this growth hormone-releasing factor (GRF) on GH release and the GH mRNA levels in monolayer c...

  5. Application of simple fed-batch technique to high-level secretory production of insulin precursor using Pichia pastoris with subsequent purification and conversion to human insulin

    Directory of Open Access Journals (Sweden)

    Chugh Dipti


    Full Text Available Abstract Background The prevalence of diabetes is predicted to rise significantly in the coming decades. A recent analysis projects that by the year 2030 there will be ~366 million diabetics around the world, leading to an increased demand for inexpensive insulin to make this life-saving drug also affordable for resource poor countries. Results A synthetic insulin precursor (IP-encoding gene, codon-optimized for expression in P. pastoris, was cloned in frame with the Saccharomyces cerevisiae α-factor secretory signal and integrated into the genome of P. pastoris strain X-33. The strain was grown to high-cell density in a batch procedure using a defined medium with low salt and high glycerol concentrations. Following batch growth, production of IP was carried out at methanol concentrations of 2 g L-1, which were kept constant throughout the remaining production phase. This robust feeding strategy led to the secretion of ~3 gram IP per liter of culture broth (corresponding to almost 4 gram IP per liter of cell-free culture supernatant. Using immobilized metal ion affinity chromatography (IMAC as a novel approach for IP purification, 95% of the secreted product was recovered with a purity of 96% from the clarified culture supernatant. Finally, the purified IP was trypsin digested, transpeptidated, deprotected and further purified leading to ~1.5 g of 99% pure recombinant human insulin per liter of culture broth. Conclusions A simple two-phase cultivation process composed of a glycerol batch and a constant methanol fed-batch phase recently developed for the intracellular production of the Hepatitis B surface antigen was adapted to secretory IP production. Compared to the highest previously reported value, this approach resulted in an ~2 fold enhancement of IP production using Pichia based expression systems, thus significantly increasing the efficiency of insulin manufacture.

  6. Screening and assessment of performance and molecule quality attributes of industrial cell lines across different fed-batch systems. (United States)

    Rouiller, Yolande; Bielser, Jean-Marc; Brühlmann, David; Jordan, Martin; Broly, Hervé; Stettler, Matthieu


    The major challenge in the selection process of recombinant cell lines for the production of biologics is the choice, early in development, of a clonal cell line presenting a high productivity and optimal cell growth. Most importantly, the selected candidate needs to generate a product quality profile which is adequate with respect to safety and efficacy and which is preserved across cell culture scales. We developed a high-throughput screening and selection strategy of recombinant cell lines, based on their productivity in shaking 96-deepwell plates operated in fed-batch mode, which enables the identification of cell lines maintaining their high productivity at larger scales. Twelve recombinant cell lines expressing the same antibody with different productivities were selected out of 470 clonal cell lines in 96-deepwell plate fed-batch culture. They were tested under the same conditions in 50 mL vented shake tubes, microscale and lab-scale bioreactors in order to confirm the maintenance of their performance at larger scales. The use of a feeding protocol and culture conditions which are essentially the same across the different scales was essential to maintain productivity and product quality profiles across scales. Compared to currently used approaches, this strategy has the advantage of speeding up the selection process and increases the number of screened clones for getting high-producing recombinant cell lines at manufacturing scale with the desired performance and quality.

  7. Specific Growth Rate Determines the Sensitivity of Escherichia coli to Lactic Acid Stress: Implications for Predictive Microbiology

    Directory of Open Access Journals (Sweden)

    Roland Lindqvist


    Full Text Available This study tested the hypothesis that sensitivity of Escherichia coli to lactic acid at concentrations relevant for fermented sausages (pH 4.6, 150 mM lactic acid, aw=0.92, temperature = 20 or 27°C increases with increasing growth rate. For E. coli strain 683 cultured in TSB in chemostat or batch, subsequent inactivation rates when exposed to lactic acid stress increased with increasing growth rate at harvest. A linear relationship between growth rate at harvest and inactivation rate was found to describe both batch and chemostat cultures. The maximum difference in T90, the estimated times for a one-log reduction, was 10 hours between bacteria harvested during the first 3 hours of batch culture, that is, at different growth rates. A 10-hour difference in T90 would correspond to measuring inactivation at 33°C or 45°C instead of 37°C based on relationships between temperature and inactivation. At similar harvest growth rates, inactivation rates were lower for bacteria cultured at 37°C than at 15–20°C. As demonstrated for E. coli 683, culture conditions leading to variable growth rates may contribute to variable lactic acid inactivation rates. Findings emphasize the use and reporting of standardised culture conditions and can have implications for the interpretation of data when developing inactivation models.

  8. Automated detection of micro-organisms in blood cultures by means of the Malthus Microbiological Growth Analyser. (United States)

    Brown, D F; Warner, M; Taylor, C E; Warren, R E


    A prototype Malthus Microbiological Growth Analyser was compared with conventional methods for examining blood cultures in a trial of 651 cultures mostly from patients with haematological malignancy or undergoing haemodialysis or renal transplantation. Of 100 significantly positive cultures, organisms from 82 grew in the conventional aerobic (+ CO2) bottle, 78 in the conventional anaerobic bottle and 71 in the Malthus bottle. The differences were not statistically significant (p greater than 0.05). The Malthus system detected 83.6% of significantly positive cultures earlier than the comparable conventional bottles while 7.3% positive cultures were detected earlier by the conventional system. When use of the Malthus system was restricted to the hours of 09.00 to 17.30 daily 27.3% positive cultures were detected earlier by the Malthus system and 16.4% were detected earlier by the conventional system. One of the organisms which grew in the Malthus bottle, a contaminating Staphylococcus epidermidis, was not detected by the Malthus system. Instability of electrodes resulted in 26.9% false positive cultures with the prototype Malthus system. Contamination rates in both the Malthus and conventional anaerobic bottles were lower than in the aerobic bottles.

  9. Polyamine levels as related to growth, differentiation and senescence in protoplast-derived cultures of Vigna aconitifolia and Avena sativa (United States)

    Kaur Sawhney, R.; Shekhawat, N. S.; Galston, A. W.


    We have previously reported that aseptically cultured mesophyll protoplasts of Vigna divide rapidly and regenerate into complete plants, while mesophyll protoplasts of Avena divide only sporadically and senesce rapidly after isolation. We measured polyamine titers in such cultures of Vigna and Avena, to study possible correlations between polyamines and cellular behavior. We also deliberately altered polyamine titer by the use of selective inhibitors of polyamine biosynthesis, noting the effects on internal polyamine titer, cell division activity and regenerative events. In Vigna cultures, levels of free and bound putrescine and spermidine increased dramatically as cell division and differentiation progressed. The increase in bound polyamines was largest in embryoid-forming callus tissue while free polyamine titer was highest in root-forming callus. In Avena cultures, the levels of total polyamines decreased as the protoplast senesced. The presence of the inhibitors alpha-difluoromethyl-arginine (specific inhibitor of arginine decarboxylase), alpha-difluoromethylornithine (specific inhibitor of ornithine decarboxylase) and dicyclohexylamine (inhibitor of spermidine synthase) reduced cell division and organogenesis in Vigna cultures. Addition of low concentration of polyamines to such cultures containing inhibitors or removal of inhibitors from the culture medium restored the progress of growth and differentiation with concomitant increase in polyamine levels.

  10. In vitro production of a caprine embryo from a preantral follicle cultured in media supplemented with growth hormone. (United States)

    Magalhães, D M; Duarte, A B G; Araújo, V R; Brito, I R; Soares, T G; Lima, I M T; Lopes, C A P; Campello, C C; Rodrigues, A P R; Figueiredo, J R


    The objective was to evaluate the effects of growth hormone (GH) on the survival, growth, maturation, and fertilization of oocytes derived from caprine preantral ovarian follicles cultured in vitro. Preantral follicles were isolated from the cortex of caprine ovaries and individually cultured for 18 d in the absence (control) or presence of bovine GH at concentrations of 10 or 50 ng/mL (GH10 and GH50, respectively). Follicle development was evaluated on the basis of survival, antral cavity formation, diameter increase, and the presence of healthy cumulus-oocyte complexes and mature oocytes. After culture, oocytes were subjected to in vitro maturation (IVM) and in vitro fertilization (IVF). The rate of antrum formation after Day 6 of culture was higher in both GH10 and GH50 than in the control (81.0, 92.7, and 47.6%, respectively, P produced mature oocytes, and enabled production of an embryo after IVF than in the control group (0.0%; P growth and maturation of goat preantral follicle oocytes and enabled production of an embryo. Furthermore, this study was apparently the first to produce a caprine embryo by in vitro fertilization of oocytes derived from preantral follicles grown in vitro.

  11. Growth rate analysis and protein identification of Kappaphycus alvarezii (Rhodophyta, Gigartinales under pH induced stress culture

    Directory of Open Access Journals (Sweden)

    Mian Zi Tee


    Full Text Available Environmental pH is one of the factors contributing to abiotic stress which in turn influences the growth and development of macroalgae. This study was conducted in order to assess the growth and physiological changes in Kappaphycus alvarezii under different pH conditions: pHs 6, ∼8.4 (control and 9. K. alvarezii explants exhibited a difference in the daily growth rate (DGR among the different pH treatments (p ≤ 0.05. The highest DGR was observed in control culture with pH ∼8.4 followed by alkaline (pH 9 and acidic (pH 6 induced stress cultures. Protein expression profile was generated from different pH induced K. alvarezii cultures using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE followed by protein identification and analysis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF-MS and Mascot software. Ribulose bisphosphate carboxylase (Rubisco large chain was identified to be up-regulated under acidic (pH 6 condition during the second and fourth week of culture. The findings indicated that Rubisco can be employed as a biomarker for pH induced abiotic stress. Further study on the association between the expression levels of Rubisco large chain and their underlying mechanisms under pH stress conditions is recommended.

  12. Mitogenic properties of insulin-like growth factors I and II, insulin-like growth factor binding protein-3 and epidermal growth factor on human breast stromal cells in primary culture. (United States)

    Strange, Karen S; Wilkinson, Darcy; Edin, Glenn; Emerman, Joanne T


    Insulin-like growth factors I and II (IGF-I and IGF-II) are growth factors implicated in both normal mammary gland development and breast cancer. We have previously reported on the effects of components of the IGF system on breast epithelial cells. Since data suggests that stromal-epithelial interactions play a crucial role in breast cancer, we have now investigated the mitogenic properties of IGF-I, IGF-II, insulin-like growth factor binding protein-3 (IGFBP-3) and epidermal growth factor (EGF) on human breast stromal cells in primary culture. We show that, under serum-free conditions, stromal cells are stimulated to grow in response to IGF-I and IGF-II in a dose-dependent manner. IGF-I and EGF, a potent stimulator of human breast epithelial cell growth in primary culture and also associated with breast cancer, appear to stimulate stromal cell growth in a synergistic manner. IGFBP-3 does not inhibit the stimulation of growth by IGF-I, or IGF-I plus EGF. However, IGFBP-3 does inhibit the stimulation of growth by IGF-II. In contrast to our previous results with human breast epithelial cells, IGFBP-3 does not have an IGF-independent inhibitory effect on stromal cell growth. This study is the first to address the effects of IGF-I, IGF-II and IGFBP-3 alone and in combination with EGF on human breast stromal cell growth in primary culture. Characterizing the role of the IGF system in both normal breast epithelial cells and stromal cells will aid in our understanding of the mechanisms behind the role of the IGF system in breast cancer.

  13. Counselor Development in the Process of Mastering Cultural Competence: A Study of Professional Growth Experiences (United States)

    Wakefield, Marie A.


    Grounded theory methodology was employed to explore the experiences of counseling professionals as they work to develop a higher level of cultural competence. Three key findings support the core theme, navigating change toward cultural competent practices: (1) environmental awareness; (2) dispositions toward the development in cultural competency…

  14. Influence of different species of Penicillium and their culture filtrates on seed germination and seedling growth of sorghum

    Directory of Open Access Journals (Sweden)

    Koteswara Rao Vankudoth


    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 The present investigations influence of Penicillium species and their culture filtrates on the seed germination and seedling growth of sorghum was studied. Culture filtrate of P. nordicum, P. verrucosum, P. chrysogenum, P. commune, P. citrinum, P.camemberti and P. digitatum were comparatively more toxic and significantly inhibited the seed germination and seedling growth varied with the species. However, pathogenicity of different inoculum of Penicillium species also exhibited varying seedling growth inhibition with P. citrinum, P. chrysogenum, P. commune, P. italicum, P. verrucosum and P. expansum assayed by water agar method. A significant phytotoxicity of these species inhibited coleoptile 32-70%, radicle 10-87% and leaf growth 20-86% with correlation coefficient 0.65, 0.67 and 0.79%, and were observed respectively. In-vitro mycotoxin production was assayed by culture filtrates of major mycotoxigenic strains revealed production of ochratoxin A (OTA, cyclopiazonic acid (CPA, rubratoxin B (RTB, griseofulvin (GRI, citrinin (CIT, patulin (PAT, penitrem A (PENA and mycophenolic acid (MPA screened by TLC/HPLC. Toxicity of species of Penicillium on seed germination, coleoptile, radicle, and leaf elongation inhibition may be attributed to the toxin-chemotypes produced by the species of Penicillium. However, even non-toxigenic strains of Penicillium also caused mild inhibition which may be attributed to the presence of other toxin-chemotypes.

  15. The chalcone butein from Rhus verniciflua Stokes inhibits clonogenic growth of human breast cancer cells co-cultured with fibroblasts

    Directory of Open Access Journals (Sweden)

    Tan Jenny


    Full Text Available Abstract Background Butein (3,4,2',4'-tetrahydroxychalone, a plant polyphenol, is a major biologically active component of the stems of Rhus verniciflua Stokes. It has long been used as a food additive in Korea and as an herbal medicine throughout Asia. Recently, butein has been shown to suppress the functions of fibroblasts. Because fibroblasts are believed to play an important role in promoting the growth of breast cancer cells, we investigated the ability of butein to inhibit the clonogenic growth of small numbers of breast cancer cells co-cultured with fibroblasts in vitro. Methods We first measured the clonogenic growth of small numbers of the UACC-812 human breast cancer cell line co-cultured on monolayers of serum-activated, human fibroblasts in the presence of butein (2 μg/mL or various other modulators of fibroblast function (troglitazone-1 μg/mL; GW9662-1 μM; meloxican-1 μM; and 3,4 dehydroproline-10 μg/mL. In a subsequent experiment, we measured the dose-response effect on the clonogenic growth of UACC-812 breast cancer cells by pre-incubating the fibroblasts with varying concentrations of butein (10 μg/ml-1.25 μg/mL. Finally, we measured the clonogenic growth of primary breast cancer cells obtained from 5 clinical specimens with normal fibroblasts and with fibroblasts that had been pre-treated with a fixed dose of butein (2.5 μg/mL. Results Of the five modulators of fibroblast function that we tested, butein was by far the most potent inhibitor of clonogenic growth of UACC-812 breast cancer cells co-cultured with fibroblasts. Pre-treatment of fibroblasts with concentrations of butein as low as 2.5 μg/mL nearly abolished subsequent clonogenic growth of UACC-812 breast cancer cells co-cultured with the fibroblasts. A similar dose of butein had no effect on the clonogenic growth of breast cancer cells cultured in the absence of fibroblasts. Significantly, clonogenic growth of the primary breast cancer cells was also

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

    Directory of Open Access Journals (Sweden)

    José Soares do Nascimento


    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

  17. Feasibility, density and growth of culture of freshwater crayfish (Decapoda: Cherax Quadricarinatus in Sinaloa, México

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    Juan Francisco Arzola González


    Full Text Available Feasibility, density and growth of culture of freshwater crayfish (Cherax quadricarinatus in Sinaloa were analyzed between September, 2005 and January, 2006. The densities were 7 (reservoir 1 and 16 (reservoir 2 org/m-2. Applied a method not parametrico (Kruskal-Wallis, the variables of response were density, weight and sex, the experimental unit the reservoirs. Samplings were weekly and 40 lobsters were captured in tubes of PVC. Total and cefalotorácica length ((LT y LC in mm and total weight (PT in g of the organisms were recorded, the biometric correlations were realized and von Bertalanffy estimated the growth for the model of von Bertallanfy. The major average was obtained in 41.2 g to a density of 7 org/m, though her k (growth rate coincided with the reservoirs with 0.30, in addition, the lobsters they presented a growth of isometric type. From the first year, usefulness of $3,419,165.00 obtained, that represented 26.8 % of the total of income, keeping a growth like that supported until the fifth year (2011. The financial evaluation determined that the investment is profitable economically, his analysis of VAN was $9,060,554.00, and in agreement to the financial indicators, this value was ideal for the accomplishment of the project. As indicator of the profitability, the TIR resulted in 66 %, being major that the minimal acceptable rate of performance and concluding the profitability of the culture in 59 %. Likewise, Sinaloa's condition, for his geographical location and the results ofthe growth, the heights, the performance of the lobster and of the financial analysis of the culture, indicated to C. quadricarinatusas an alternative of development in the region, being the lobster of interest in acuiculture of new species in tropical areas, thoughin Sinaloa the investigations are practically void with ends of commercial production.

  18. Arbuscular mycorrhizae enhance metal lead uptake and growth of host plants under a sand culture experiment. (United States)

    Chen, Xin; Wu, Chunhua; Tang, Jianjun; Hu, Shuijin


    A sand culture experiment was conducted to investigate whether mycorrhizal colonization and mycorrhizal fungal vesicular numbers were influenced by metal lead, and whether mycorrhizae enhance host plants tolerance to metal lead. Metal lead was applied as Pb(NO3)2 in solution at three levels (0, 300 and 600 mg kg(-1) sand). Five mycorrhizal host plant species, Kummerowia striata (Thunb.) Schindl, Ixeris denticulate L., Lolium perenne L., Trifolium repens L. and Echinochloa crusgalli var. mitis were used to examine Pb-mycorrhizal interactions. The arbuscular mycorrhizal inoculum consisted of mixed spores of mycorrhizal fungal species directly isolated from orchard soil. Compared to the untreated control, both Pb concentrations reduced mycorrhizal colonization by 3.8-70.4%. Numbers of AM fungal vesicles increased by 13.2-51.5% in 300 mg Pb kg(-1) sand but decreased by 9.4-50.9% in 600 mg Pb kg(-1) sand. Mycorrhizae significantly enhanced Pb accumulation both in shoot by 10.2-85.5% and in root by 9.3-118.4%. Mycorrhizae also enhanced shoot biomass and shoot P concentration under both Pb concentrations. Root/shoot ratios of Pb concentration were higher in highly mycorrhizal plant species (K.striata, I. denticulate, and E. crusgalli var. mitis) than that in poorly mycorrhizal ones (L. perenne and T. repens,). Mycorrhizal inoculation increased the root/shoot ratio of Pb concentration of highly mycorrhizal plant species by 7.6-57.2% but did not affect the poorly mycorrhizal ones. In the treatments with 300 Pb mg kg(-1) sand, plant species with higher vesicular numbers tended to show higher root/shoot ratios of the Pb concentration. We suggest that under an elevated Pb condition, mycorrhizae could promote plant growth by increasing P uptake and mitigate Pb toxicity by sequestrating more Pb in roots.

  19. Insulin receptors mediate growth effects in cultured fetal neurons. I. Rapid stimulation of protein synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Heidenreich, K.A.; Toledo, S.P. (Univ. of California-San Diego, La Jolla (USA))


    In this study we have examined the effects of insulin on protein synthesis in cultured fetal chick neurons. Protein synthesis was monitored by measuring the incorporation of (3H)leucine (3H-leu) into trichloroacetic acid (TCA)-precipitable protein. Upon addition of 3H-leu, there was a 5-min lag before radioactivity occurred in protein. During this period cell-associated radioactivity reached equilibrium and was totally recovered in the TCA-soluble fraction. After 5 min, the incorporation of 3H-leu into protein was linear for 2 h and was inhibited (98%) by the inclusion of 10 micrograms/ml cycloheximide. After 24 h of serum deprivation, insulin increased 3H-leu incorporation into protein by approximately 2-fold. The stimulation of protein synthesis by insulin was dose dependent (ED50 = 70 pM) and seen within 30 min. Proinsulin was approximately 10-fold less potent than insulin on a molar basis in stimulating neuronal protein synthesis. Insulin had no effect on the TCA-soluble fraction of 3H-leu at any time and did not influence the uptake of (3H)aminoisobutyric acid into neurons. The isotope ratio of 3H-leu/14C-leu in the leucyl tRNA pool was the same in control and insulin-treated neurons. Analysis of newly synthesized proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that insulin uniformly increased the incorporation of 14C-leu into all of the resolved neuronal proteins. We conclude from these data that (1) insulin rapidly stimulates overall protein synthesis in fetal neurons independent of amino acid uptake and aminoacyl tRNA precursor pools; (2) stimulation of protein synthesis is mediated by the brain subtype of insulin receptor; and (3) insulin is potentially an important in vivo growth factor for fetal central nervous system neurons.

  20. Influence of nickel nutrition and nitrogen source on growth and yield of lettuce in hydroponic culture

    Directory of Open Access Journals (Sweden)

    F. Hosseini


    Full Text Available Nickel (Ni is the most recently discovered essential element for higher plants. But there is limited information about the effect of this element on yield and nitrogen (N metabolism of different plants. In this research, the interaction of Ni supplement and N source was studied on nitrate accumulation and growth of lettuce (Lactuca sativa L. cv. Baker in solution culture. In a greenhouse experiment, lettuce plants were exposed to two Ni levels (0 and 0.04 µM in the form of NiCl2 and three N levels (5, 10, and 20 µM in the form of urea and ammonium nitrate. The plants were harvested 6 weeks after transplanting and the fresh weight of shoots and roots, total N concentration, nitrate concentration and Fe content of shoots were determined. The results indicated that shoot and root fresh weight of nitrate-fed plants were higher than urea-fed plants. Ni nutrition significantly enhanced the yield of urea-fed plants. The shoot nitrate concentration in nitrate-fed plants was significantly greater in comparison with urea-fed plants. On the other hand, nitrate concentration in the shoots of lettuce plants decreased significantly with Ni nutrition. Ni supplementation in urea-fed plants increased the shoot Fe content. The shoot concentration of total-N in the urea-fed plants increased with the Ni supplement. While, Ni had no significant effect on shoot concentration of total-N in the nitrate-fed plants. Addition of low levels of Ni to the nutrient solution, particularly to the urea-containing solution, improved the yield of lettuce. In addition, these plant leaves are safer for human consumption because the shoot nitrate content is significantly reduced by application of Ni.

  1. Increase in the Synthesis of Polyfructan in the Cultures of Chicory “Hairy Roots” with Plant Natural Growth Regulators

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    Victoria А. Tsygankova, PhD¹


    Full Text Available Experiments were conducted to study the benefit of using the new plant growth regulators (PGRs Ivin, Emistim, Biolan and Charkor in nutrient medium ½ MS for intensification of biomass growth and the increased synthesis of polyfructan (PF in the cultures of chicory “hairy roots” (Cichorium intybus L., obtained by Аgrobacterium rhizogenes-mediated transformation,. The best indexes of increased specific quantities of PF are observed after using Biolan at a concentration of 5.0 μL/L (up to 130 mg/g dry mass of roots and Emistim at a concentration 2.5 µL/L (up to 220 mg/g dry mass of roots. The greatest stimulation of root growth activity was expressed on using the growth regulators Emistim, Ivin and Charkor, in concentrations between 2.5 and 10.0 µL/L, considerably raising the total quantity of PF: compared with the control the use of regulator Emistim showed a rise of up to 35 times, regulator Ivin showed an increase of up to 28 times and regulator Charkor showed an increase up to 7.0-7.5 times. The results thus obtained definitely prove the benefit of applying these regulators to increase the biomass growth and PF synthesis in the culture of chicory «hairy roots».

  2. Fed-batch cultivation of Arthrospira and Chlorella in ammonia-rich wastewater: Optimization of nutrient removal and biomass production. (United States)

    Markou, Giorgos


    In the present work the cyanobacterium Arthrospira platensis and the microalga Chlorella vulgaris were fed-batch cultivated in ammonia-rich wastewater derived from the anaerobic digestion of poultry litter. Aim of the study was to maximize the biomass production along with the nutrient removal aiming to wastewater treatment. Ammonia and phosphorus removals were very high (>95%) for all cultures investigated. Both microorganisms were able to remove volatile fatty acids to an extent of >90%, indicating that they were capable of mixotrophic growth. Chemical oxygen demand and proteins were also removed in various degrees. In contrast, in all cultures carbohydrate concentration was increased. The biochemical composition of the microorganisms varied greatly and was influenced by the indicate that the nutrient availability. A. platensis accumulated carbohydrates (≈ 40%), while C. vulgaris accumulated lipids (≈ 50%), rendering them interesting for biofuel production.

  3. Leishmania (Viannia braziliensis growth in vitro culture relies more on folic acid availability than Leihsmania (Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Andrea Niño


    Full Text Available We compared the in vitro growth of promastigotes from two Leishmania species in TC-100 and Schneider media. Leishmania (Leishmania amazonensis replication rates were similar in both tissue culture media and reached maximum rates by 48 h. In contrast Leishmania (Viannia braziliensis growth was significantly greater in TC-100 but maximum rates were achieved by 96 h. Folic acid appears to be the limiting factor and supplementation of Schneider media with this nutrient improved L. (V. braziliensis replication rates and decreased the time of maximum replication to 48 h.

  4. Increase in the Synthesis of Polyfructan in the Cultures of Chicory “Hairy Roots” with Plant Natural Growth Regulators


    Victoria А. Tsygankova, PhD¹; Alla I. Yemets, PhD, ScD²; Sergey P. Ponomarenko, PhD, ScD³; Nadezhda A. Matvieieva, PhD; Sergey E. Chapkevich"; Nikolay V. Kuchuk, PhD, ScD"


    Experiments were conducted to study the benefit of using the new plant growth regulators (PGRs) Ivin, Emistim, Biolan and Charkor in nutrient medium ½ MS for intensification of biomass growth and the increased synthesis of polyfructan (PF) in the cultures of chicory “hairy roots” (Cichorium intybus L.), obtained by Аgrobacterium rhizogenes-mediated transformation,. The best indexes of increased specific quantities of PF are observed after using Biolan at a concentration of 5.0 μL/L (up to 130...

  5. Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture. (United States)

    Imsoonthornruksa, Sumeth; Pruksananonda, Kamthorn; Parnpai, Rangsun; Rungsiwiwut, Ruttachuk; Ketudat-Cairns, Mariena


    To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins.

  6. Quick adaptation of Ralstonia Solanacearum to copper stress to recover culturability and growth in water and soil

    Directory of Open Access Journals (Sweden)

    Sergio Daniel Moreira Ascarrunz


    Full Text Available Cells of Ralstonia solanacearum were exposed to Cu in distilled water, and the resulting Cu-stressed non-culturable cells were inoculated to natural (non-pasteurized and pasteurized soils in order to examine their culturability and recovery. Exposing the cells to 20 µM CuSO4 produced transitory non-culturable cells, which exhibited a remarkable recovery in culturability after incubation in the solution for 36 h, reaching a density near the initial level by 108 h. To determine whether such non-culturable cells actually "resuscitated" or multiplied after adapting to Cu toxicity, growth curves were constructed in order to contrast the rates of increase in culturable cell numbers between Cu-stressed or non-stressed inocula. Additionally, fresh non-stressed cells were exposed to CuSO4 in the presence of nalidixic acid by adding the antibiotic at different times after the onset of Cu stress to verify any cell multiplication during the population increase. The results revealed that the non-culturable cells surviving Cu toxicity adapted very quickly to Cu and began multiplying within 12 h, because only the Cu-stressed cells that were increasing in the exponential growth phase, but not those in the stationary phase, were killed by the antibiotic. Such cells exhibited an apparent tolerance to this metal when inoculated to a freshly prepared solution of CuSO4, and also detoxified the ion in the solution in which they grew. The presence of nutrients greatly counteracted the effect of Cu in water microcosms, since culturable cells were detected and increased in number even when exposed to 40 µM CuSO4. In contrast, when inoculated to non-pasteurized soil, Cu-stressed cells showed no such recoveries. However, when the soil was pasteurized before inoculation or added with nutrients, culturable cells were recovered and increased in number. This indicates that increased nutrient availability in soil allows Cu-stressed cells to quickly overcome the stress and

  7. Identifying conditions for inducible protein production in E. coli: combining a fed-batch and multiple induction approach

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    Choi Young J


    Full Text Available Abstract Background In the interest of generating large amounts of recombinant protein, inducible systems have been studied to maximize both the growth of the culture and the production of foreign proteins. Even though thermo-inducible systems were developed in the late 1970's, the number of studies that focus on strategies for the implementation at bioreactor scale is limited. In this work, the bacteriophage lambda PL promoter is once again investigated as an inducible element but for the production of green fluorescent protein (GFP. Culture temperature, induction point, induction duration and number of inductions were considered as factors to maximize GFP production in a 20-L bioreactor. Results It was found that cultures carried out at 37°C resulted in a growth-associated production of GFP without the need of an induction at 42°C. Specific production was similar to what was achieved when separating the growth and production phases. Shake flask cultures were used to screen for desirable operating conditions. It was found that multiple inductions increased the production of GFP. Induction decreased the growth rate and substrate yield coefficients; therefore, two time domains (before and after induction having different kinetic parameters were created to fit a model to the data collected. Conclusion Based on two batch runs and the simulation of culture dynamics, a pre-defined feeding and induction strategy was developed to increase the volumetric yield of a temperature regulated expression system and was successfully implemented in a 20-L bioreactor. An overall cell density of 5.95 g DW l-1 was achieved without detriment to the cell specific production of GFP; however, the production of GFP was underestimated in the simulations due to a significant contribution of non-growth associated product formation under limiting nutrient conditions.

  8. Growth Characteristics of Pearl Oyster Pinctada Maxiam Seeds with Broodstocks Introduced from Indonesia Cultured in the Xuwen Sea of Zhanjiang%广东徐闻海域印尼产大珠母贝的生长特性

    Institute of Scientific and Technical Information of China (English)

    梁飞龙; 谢绍河; 符韶; 邓岳文


    为将印尼产的大珠母贝作为我国海水珍珠养殖的新品种,分别于2010年6月4日、7月20日和10月28日,利用从印度尼西亚引进的大珠母贝作为亲本培育了3批苗种,在广东湛江徐闻承梧海区进行了养殖试验,实验期间水温14~32℃,盐度28.4~36.9.结果表明:除了2010年10月28日出池的第3批苗,养殖11个月,成活率为1.3±0.5%外.经过12个月养殖,第一、二批苗的贝体平均壳高、壳长、体质量和成活率分别为59.56±10.2mm,60.39±10.5mm,25.62±6.5g,10.6%±1.8%和54.84±6.1mm,55.96±12.4mm,18.40±6.1g,25.10%±2.6%.%During the period of June 2010 to September 2011, three batches of pearl oyster Pinctada maxima were obtained with breeder introduced from Indonesia and then cultured in the Chengwu sea area of Xuwen, Zhanjiang City of Guangdong Province. Growth of the three groups was evaluated on the basis of shell height, shell length and total weight measurement at ages. During the experimental period, temperature was 14.0-32.0℃ and salinity of seawater was 28.4-36.9, respectively. Except for the third batch spat of pearl oyster Pinctada maxima, which started to be cultured at low temperature in winter, the survival rate was 1.3%±0.5% after culture for llmonths. After culturing in the sea area for 12 months, the mean value of shell height, shell length , body weight and survival rate of the first and second batch spat was 59.56 mm, 60.39 mm, 25.62 g and 10.6%; 54.84 mm, 55.96 mm, 18.40 g and 25.10%, respectively. The results showed that low temperature in winter had significant effect on the growth rate and survival rate of small oysters of P. Maxima.

  9. In vitro propagation of plant virus using different forms of plant tissue culture and modes of culture operation. (United States)

    Shih, Sharon M-H; Doran, Pauline M


    Plant virus accumulation was investigated in vitro using three different forms of plant tissue culture. Suspended cells, hairy roots and shooty teratomas of Nicotiana benthamiana were infected with tobacco mosaic virus (TMV) using the same initial virus:biomass ratio. Viral infection did not affect tissue growth or morphology in any of the three culture systems. Average maximum virus concentrations in hairy roots and shooty teratomas were similar and about an order of magnitude higher than in suspended cells. Hairy roots were considered the preferred host because of their morphological stability in liquid medium and relative ease of culture. The average maximum virus concentration in the hairy roots was 0.82+/-0.14 mg g(-1) dry weight; viral coat protein represented a maximum of approximately 6% of total soluble protein in the biomass. Virus accumulation in hairy roots was investigated further using different modes of semi-continuous culture operation aimed at prolonging the root growth phase and providing nutrient supplementation; however, virus concentrations in the roots were not enhanced compared with simple batch culture. The relative infectivity of virus in the biomass declined by 80-90% during all the cultures tested, irrespective of the form of plant tissue used or mode of culture operation. Hairy root cultures inoculated with a transgenic TMV-based vector in batch culture accumulated green fluorescent protein (GFP); however, maximum GFP concentrations in the biomass were relatively low at 39 microg g(-1) dry weight, probably due to genetic instability of the vector. This work highlights the advantages of using hairy roots for in vitro propagation of TMV compared with shooty teratomas and suspended plant cells, and demonstrates that batch root culture is more effective than semi-continuous operations for accumulation of high virus concentrations in the biomass.

  10. The Research of Growth Capability Intensification and Lipolytic Activity of Fat Splitting Microbial Cultures by Influence of Natural Mineral Components

    Directory of Open Access Journals (Sweden)

    Olga V. Kolotova


    Full Text Available 23 bacterial strains with lipase activity were separated from the waste water of the meat-processing enterprises and Volgograd water-handling facilities aeration tank. There were studied their morphological, cultural and biochemical properties and their animal and vegetable fat splitting ability. Comparative analysis of fat splitting ability has been carried out for searched strains and devices of enzyme activity increasing and growing process intensity of searched microorganisms were investigated. Mineral nature ingredients – the lake Elton brine, the bischofite, the Dead Sea salt – were added in the culture mediums for increasing of biomass accumulation. It has been investigated that addition of the brine into the culture medium in 5 % concentration increases the growth rate of producer lipase on 293 % in comparison with growth of the control medium without the brine. Addition of 0,25 % the Dead Sea salt concentration intensifies the growth on 317 % in comparison with the control medium. Laboratory simulation of fat waste water purification process has been realized with one of separated activity bacterial strains and it has been showed that addition of the 0,5 % bishofite into the fermenter stimulates lipase activity of the searched microorganisms.

  11. The role of basic fibroblast growth factor in glioblastoma multiforme and glioblastoma stem cells and in their in vitro culture. (United States)

    Haley, Elizabeth M; Kim, Yonghyun


    Glioblastoma multiforme (GBM) is the most malignant form of central nervous system tumor, and current therapies are largely ineffective at treating the cancer. Developing a more complete understanding of the mechanisms controlling the tumor is important in order to explore new possible treatment options. It is speculated that the presence of glioblastoma stem or stem-like cells (GSCs), a rare type of pluripotent cancer cell that possesses the ability to self-renew and generate tumors, could be an important factor contributing to the resistance to treatment and deadliness of the cancer. A comprehensive knowledge of the mechanisms controlling the expression and properties of GSCs is currently lacking, and one promising area for further exploration is in the influence of basic fibroblast growth factor (FGF-2) on GSCs. Recent studies reveal that FGF-2 plays a significant part in regulating GBM, and the growth factor is commonly included as a supplement in media used to culture GSCs in vitro. However, the particular role that FGF-2 plays in GSCs has not been as extensively explored. Therefore, understanding how FGF-2 is involved in GSCs and in GBMs could be an important step towards a more complete comprehension of the managing the disease. In this review, we look at the structure, signaling pathways, and specific role of FGF-2 in GBM and GSCs. In addition, we explore the use of FGF-2 in cell culture and using its synthetic analogs as a potential alternative to the growth factor in culture medium.

  12. Cross-scale predictive modeling of CHO cell culture growth and metabolites using Raman spectroscopy and multivariate analysis. (United States)

    Berry, Brandon; Moretto, Justin; Matthews, Thomas; Smelko, John; Wiltberger, Kelly


    Multi-component, multi-scale Raman spectroscopy modeling results from a monoclonal antibody producing CHO cell culture process including data from two development scales (3 L, 200 L) and a clinical manufacturing scale environment (2,000 L) are presented. Multivariate analysis principles are a critical component to partial least squares (PLS) modeling but can quickly turn into an overly iterative process, thus a simplified protocol is proposed for addressing necessary steps including spectral preprocessing, spectral region selection, and outlier removal to create models exclusively from cell culture process data without the inclusion of spectral data from chemically defined nutrient solutions or targeted component spiking studies. An array of single-scale and combination-scale modeling iterations were generated to evaluate technology capabilities and model scalability. Analysis of prediction errors across models suggests that glucose, lactate, and osmolality are well modeled. Model strength was confirmed via predictive validation and by examining performance similarity across single-scale and combination-scale models. Additionally, accurate predictive models were attained in most cases for viable cell density and total cell density; however, these components exhibited some scale-dependencies that hindered model quality in cross-scale predictions where only development data was used in calibration. Glutamate and ammonium models were also able to achieve accurate predictions in most cases. However, there are differences in the absolute concentration ranges of these components across the datasets of individual bioreactor scales. Thus, glutamate and ammonium PLS models were forced to extrapolate in cases where models were derived from small scale data only but used in cross-scale applications predicting against manufacturing scale batches.

  13. Effects of extracellular matrix proteins on macrophage differentiation, growth, and function: comparison of liquid and agar culture systems (United States)

    Armstrong, J. W.; Chapes, S. K.; Spooner, B. S. (Principal Investigator)


    Both spaceflight and skeletal unloading suppress the haematopoietic differentiation of macrophages (Sonnenfeld et al., Aviat. Space Environ. Med., 61:648-653, 1990; Armstrong et al., J. Appl. Physiol., 75:2734-2739, 1993). The mechanism behind this reduction in haematopoiesis has yet to be elucidated. However, changes in bone marrow extracellular matrix (ECM) may be involved. To further understand the role of ECM products in macrophage differentiation, we have performed experiments evaluating the effects of fibronectin, laminin, collagen type I, and collagen type IV on macrophage development and function. Bone marrow-derived macrophages cultured on four different ECM substrates in liquid culture medium showed less growth than those cultured on plastic. Significant morphological differences were seen on each of the substrates used. Phenotypically and functionally, as measured by class II major histocompatibility molecule (MHCII) expression, MAC-2 expression, and the secretion of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), these macrophages were similar. In contrast, bone marrow-derived macrophages cultured in suspension, using agar, showed no difference in growth when exposed to ECM proteins. However, IL-6 and TNF-alpha secretion was affected by fibronectin, laminin, collagen type I, and collagen type IV in a concentration-dependent manner. We conclude that the ECM products fibronectin, laminin, collagen type I, and collagen type IV have profound effects on macrophage development and function. Additionally, we suggest that an ECM-supplemented agar culture system provides an environment more analogous to in vivo bone marrow than does a traditional liquid culture system.

  14. Dynamics of algae growth and nutrients in experimental enclosures culturing bighead carp and common carp:Phosphorus dynamics

    Institute of Scientific and Technical Information of China (English)

    Suiliang Huang; Baoyan Liu; Shaoming Wang; Yang Luo; Adam Szymkiewicz; Romuald Szymkiewicz; Min Wu; Changjuan Zang; Shenglan Du; Joseph Domagalski; Magdalena Gajewska; Feng Gao; Chao Lin; Yong Guo


    This is the third paper of the series about “Dynamics of algae growth and nutrients in experimental enclosures culturing bighead carp and common carp”. In this paper, phosphorus dynamics were inves-tigated under the condition of culturing bighead carp and common carp with added fish food (nitrogen dynamics is discussed in the second paper because their behaviors are so different from each other). Nearly fifty days’ observation results indicated that the reservoir water was typical of “phosphorus limited” water, and soluble reactive phosphorus (SRP) was the main constituent of measured total phosphorus (TP). The presence of fish food resulted in significantly higher SRP, dissolved total phos-phorus (DTP) and TP concentrations in contrast with the reservoir water. Moreover, continuous supply of fish food led to the decline of total nitrogen to total phosphorus (TN:TP) from more than 100 to less than 5. Variations in the ratio of TN to TP favored the growth of blue-green algae. Fish species affected phosphorus concentrations, and culturing bait-eating common carp contributed more to reducing the SRP, DTP and TP concentrations than culturing planktivorous bighead carp. 0.5%, 4.1%and 3.1%TP can be removed in enclosures with culturing bighead carp, common carp and mixed bighead carp and common carp, respectively. Abundant phosphorus in the fish culturing activities may be present as the uneaten food, algae cells, and within the water column and sediment, which should be taken into serious con-sideration for the target of future water eutrophication prevention and safety of the drinking water supply.

  15. A candidate gene association study for growth performance in an improved giant freshwater prawn (Macrobrachium rosenbergii ) culture line. (United States)

    Jung, Hyungtaek; Lyons, Russell E; Li, Yutao; Thanh, Nguyen Minh; Dinh, Hung; Hurwood, David A; Salin, Krishna R; Mather, Peter B


    A candidate gene approach using type I single nucleotide polymorphism (SNP) markers can provide an effective method for detecting genes and gene regions that underlie phenotypic variation in adaptively significant traits. In the absence of available genomic data resources, transcriptomes were recently generated in Macrobrachium rosenbergii to identify candidate genes and markers potentially associated with growth. The characterisation of 47 candidate loci by ABI re-sequencing of four cultured and eight wild samples revealed 342 putative SNPs. Among these, 28 SNPs were selected in 23 growth-related candidate genes to genotype in 200 animals selected for improved growth performance in an experimental GFP culture line in Vietnam. The associations between SNP markers and individual growth performance were then examined. For additive and dominant effects, a total of three exonic SNPs in glycogen phosphorylase (additive), heat shock protein 90 (additive and dominant) and peroxidasin (additive), and a total of six intronic SNPs in ankyrin repeats-like protein (additive and dominant), rolling pebbles (dominant), transforming growth factor-β induced precursor (dominant), and UTP-glucose-1-phosphate uridylyltransferase 2 (dominant) genes showed significant associations with the estimated breeding values in the experimental animals (P =0.001-0.031). Individually, they explained 2.6-4.8 % of the genetic variance (R²=0.026-0.048). This is the first large set of SNP markers reported for M. rosenbergii and will be useful for confirmation of associations in other samples or culture lines as well as having applications in marker-assisted selection in future breeding programs.

  16. Transcriptional response of P. pastoris in fed-batch cultivations to Rhizopus oryzae lipase production reveals UPR induction

    Directory of Open Access Journals (Sweden)

    Valero Francisco


    Full Text Available Abstract Background The analysis of transcriptional levels of the genes involved in protein synthesis and secretion is a key factor to understand the host organism's responses to recombinant protein production, as well as their interaction with the cultivation conditions. Novel techniques such as the sandwich hybridization allow monitoring quantitatively the dynamic changes of specific RNAs. In this study, the transcriptional levels of some genes related to the unfolded protein response (UPR and central metabolism of Pichia pastoris were analysed during batch and fed-batch cultivations using an X-33-derived strain expressing a Rhizopus oryzae lipase under control of the formaldehyde dehydrogenase promoter (FLD1, namely the alcohol oxidase gene AOX1, the formaldehyde dehydrogenase FLD1, the protein disulfide isomerase PDI, the KAR2 gene coding for the BiP chaperone, the 26S rRNA and the R. oryzae lipase gene ROL. Results The transcriptional levels of the selected set of genes were first analysed in P. pastoris cells growing in shake flask cultures containing different carbon and nitrogen sources combinations, glycerol + ammonium, methanol + methylamine and sorbitol + methylamine. The transcriptional levels of the AOX1 and FLD1 genes were coherent with the known regulatory mechanism of C1 substrates in P. pastoris, whereas ROL induction lead to the up-regulation of KAR2 and PDI transcriptional levels, thus suggesting that ROL overexpression triggers the UPR. This was further confirmed in fed-batch cultivations performed at different growth rates. Transcriptional levels of the analysed set of genes were generally higher at higher growth rates. Nevertheless, when ROL was overexpressed in a strain having the UPR constitutively activated, significantly lower relative induction levels of these marker genes were detected. Conclusion The bead-based sandwich hybridization assay has shown its potential as a reliable instrument for quantification of

  17. Molecular identification and culture trials of Eutreptiella gymnastica (Eutreptiales, Euglenophyceae)

    Institute of Scientific and Technical Information of China (English)

    XU Na; PANG Shaojun; SHAN Tifeng; LIU Feng; ZHAO Xiaobo; GAO Suqin


    During a survey for marine microalgal resources,we isolated a rare marine euglenoid from the coastal waters of Qingdao,China in 2009,and established a pure culture.Electron microscopic and molecular phylogenetic (18S rDNA and 16S rDNA sequences) analyses revealed a close affinity with Eutreptiella gymnastica,a bloom-forming species.Different culture conditions were monitored to understand optimal E.gymnastica growth characteristics.The optimal growth conditions in a batch culture of this isolate were 20℃,160 μmol photons/(m2·s) of white light,and a salinity of 10-31.Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 μmol/L.Understanding the effect of culture conditions on E.gymnastica may help understanding the blooming mechanism of this alga in its natural environment.

  18. Quantification of nutrient-replete growth rates in five-ion hyperspace for Chlorella vulgaris (Trebouxiophyceae) and Peridinium cinctum (Dinophyceae). (United States)

    The effects of five ions, NO3-, PO43-, K+, Na+ and Cl- on growth rates and cell densities were quantified for Chlorella vulgaris (Chlorophycea) and Peridinium cinctum (Dinophycea) in batch cultures. A five dimensional experimental design, the five component mixture design projected across a total i...

  19. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz


    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.

  20. International WIL Placements: Their Influence on Student Professional Development, Personal Growth and Cultural Competence (United States)

    Gribble, Nigel; Dender, Alma; Lawrence, Emma; Manning, Kirrily; Falkmer, Torbjorn


    In the increasingly global world, skills in cultural competence now form part of the minimum standards of practice required for allied health professionals. During an international work-integrated learning (WIL) placement, allied health students' cultural competence is expected to be enhanced. The present study scrutinized reflective journals of…

  1. Culturally Responsive Teaching: Awareness and Professional Growth through a School-University Collaboration (United States)

    McCormick, Theresa M.; Eick, Charles J.; Womack, Janet S.


    Preparing in-service and pre-service teachers to effectively work with culturally diverse students is an ongoing challenge for schools and universities alike. This article reports on a University-Professional Development School (PDS) initiative designed to enhance an awareness of culturally responsive pedagogy. This article describes a yearlong…

  2. Impact of lactate on growth of cultures of cecal bacteria from commercial broilers (United States)

    Cultures of beneficial bacteria used in probiotics produce and utilize organic acids that may play a role in the ability of the cultures to inhibit colonization of poultry by enteropathogens. Cecal contents of adult poultry contain many of these beneficial bacteria, and earlier experiments showed th...

  3. Food additives reduce lactic acid bacterial growth in culture medium and in meat products, increasing product shelf life

    Directory of Open Access Journals (Sweden)

    Cleonice Mendes Pereira Sarmento


    Full Text Available The uncontrolled growth of lactic acid bacteria (LAB in meat and meat products leads to product spoilage, and thus shortens product shelf life. Although food additives are known to decrease LAB growth, this effect has not been analyzed in detail. Here, a detailed analysis was performed of the effects of sodium chloride, sodium polyphosphate, sodium lactate, sodium nitrite/nitrate, and garlic on the growth of the Lactobacillus plantarum in culture medium. The results were used to design and test experimental formulations of meat products. Initially, the effect of food additives on L. plantarum was evaluated using a Fractional Factorial Design (FFD, followed by a Central Composite Rotatable Design (CCRD. The Modified Gompertz Model was adjusted to the growth curves to determine the Kinetic parameters of bacterial growth (logarithmic increase in the population, specific growth rate, and lag phase extension. Higher sodium lactate and sodium chloride levels had a negative impact on L. plantarum growth parameters (p?0.05. Therefore, we designed experimental formulations of mortadella and smoked pork sausages containing 4% sodium lactate (w w-1 and 2.4-3.5% sodium chloride (w w-1, and determined LAB growth from samples of stored products produced according to these formulations, in order to determine product shelf life. There was an increased lag phase of LAB growth for most experimental formulations. Also, the experimental smoked pork sausages had a longer shelf life, which was increased by at least 22 days, suggesting that the proposed formulation, with higher than standard lactate concentration, increased the product’s shelf life.

  4. Growth characteristics and Ha-ras mutations of cell cultures isolated from chemically induced mouse liver tumours. (United States)

    Pedrick, M S; Rumsby, P C; Wright, V; Phillimore, H E; Butler, W H; Evans, J G


    Cells have been isolated from liver tumours that have arisen in control C3H/He mice, in mice given 10 micrograms diethylnitrosamine (DEN) during the neonatal period or in mice given a diet containing phenobarbitone (PB) to allow a daily intake of 85 mg/kg/day. The cells were grown to the 8 degrees subculture when their growth characteristics were investigated in monolayer culture and following suspension in soft agar and on transplantation into nude mice. In addition, DNA was isolated from the cultures and from tumours that grew in nude mice and analysed for mutations at codon 61 of the Ha-ras oncogene. All cells derived from DEN-induced hepatocellular carcinomas (HCC) demonstrated a lack of density inhibition of growth in monolayer culture, grew in soft agar and formed tumours in nude mice with an average mean latency of 29 days. Three of the seven lines showed mutations in Ha-ras: two were CAA-->AAA transversions and one showed a CAA-->CTA transversion. In contrast, cells isolated from eosinophilic nodules in mice given PB showed inhibition of growth at confluence, did not grow in soft agar and only four of eight formed tumours in nude mice with a mean average latent period of 181 days. Cells grown from HCC in mice given PB showed a lack of density inhibition of growth, however, they did not grow in soft agar nor did they form tumours in nude mice. A single spontaneous HCC from a control mouse showed a similar growth pattern to HCC cells isolated from mice given PB. Cells from a basophilic nodule, taken from a control untreated mouse grew vigorously in culture and in soft agar and formed tumours in nude mice with a latency of 6 days. None of the cells isolated from control mice or from mice given PB showed evidence of mutations at codon 61 of Ha-ras. These data confirm that there are fundamental differences in the biology of cells grown from tumours that develop in mice under different treatment regimes. These studies also demonstrate the utility of cell culture

  5. Growth of Trichomycterus punctulatus “Life” in three population densities in intensive culture system with recirculation

    Directory of Open Access Journals (Sweden)

    Segundo Juan López Cubas


    Full Text Available The present research was conducted in order to determine the effect of population density on the growth of Trichomycterus punctulatus, in an intensive culture system with recirculation, to which was applied Increasing Stimulus Experimental Design with three treatments without repetition: 2.21 lifes / liter, 2.94 lifes / liter and 3.68 lifes / liter. The fish were fed a diet of 32% protein and were sampled monthly growth as well as physical and chemical parameters of the water. We applied ANOVA and Duncan test. Growth of T. punctulatus was affected by population density in direct, with a higher density of 3.68 lifes / liter: 90.90 mm and 5.95 g; also the overall production per cubic meter and observed the same trend. The physicochemical parameters of the water were similar among treatments and were within the ranges of good development for fish.

  6. What the growth of a space tourism industry could contribute to employment, economic growth, environmental protection, education, culture and world peace (United States)

    Collins, Patrick; Autino, Adriano


    The authors argue that the creation of a popular new industry of passenger space travel could be economically and socially very beneficial in creating new employment in aerospace and related fields in order to supply these services. In doing so, the application of nearly a half-century of technological development that has yet to be used commercially could create many new aerospace engineering business opportunities. In addition, by growing to large scale, space tourism has unique potential to reduce the cost of space travel sharply, thereby making many other activities in space feasible and profitable. The paper discusses the scope for new employment, stimulating economic growth, reducing environmental damage, sustaining education particularly in the sciences, stimulating cultural growth, and preserving peace by eliminating any need for "resource wars".

  7. A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations (United States)

    Han, Jichang; Wang, Song; Zhang, Lin; Yang, Guanpin; Zhao, Lu; Pan, Kehou


    Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin (each at a concentration of 600 mg/L) for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains of Nannochloropsis sp. (Eustigmatophyceae), two strains of Cylindrotheca sp. (Bacillariophyceae), two strains of Tetraselmis sp. (Chlorodendrophyceae) and one strain of Amphikrikos sp. (Trebouxiophyceae) were purified successfully. The method shows promise for batch-purifying microalgal cultures.

  8. Growth Phase, Oxygen, Temperature and Starvation Affect the Development of Viable but Non-Culturable State of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Bin eWu


    Full Text Available AbstractVibrio cholerae can enter into a viable but non-culturable (VBNC state in order to survive in unfavourable environments. In this study, we studied the roles of five physicochemical and microbiological factors or states, namely, different strains, growth phases, oxygen, temperature, and starvation, on the development of VBNC of V. cholerae in artificial sea water (ASW. Different strains of the organism, the growth phase, and oxygen levels affected the progress of VBNC development. It was found that the VBNC state was induced faster in V. cholerae serogroup O1 classical biotype strain O395 than in O1 El Tor biotype strains C6706 and N16961. When cells in different growth phases were used for VBNC induction, stationary-phase cells lost their culturability more quickly than exponential-phase cells, while induction of a totally non-culturable state took longer to achieve for stationary-phase cells in all three strains, suggesting that heterogeneity of cells should be considered. Aeration strongly accelerated the loss of culturability. During the development of the VBNC state, the culturable cell count under aeration conditions was almost 106-fold lower than under oxygen-limited conditions for all three strains. The other two factors, temperature and nutrients-rich environment, may prevent the induction of VBNC cells. At 22°C or 37°C in ASW, most of the cells rapidly died and the culturable cell count reduced from about 108 CFU/mL to 106–105 CFU/mL. The total cell counts showed that cells that lost viability were decomposed, and the viable cell counts were the same as culturable cell counts, indicating that the cells did not reach the VBNC state. VBNC state development was blocked when ASW was supplied with Luria-Bertani broth (LB, but it was not affected in ASW with M9, suggesting that specific nutrients in LB may prevent the development of VBNC state. These results revealed that the five factors evaluated in this study had different

  9. Effects of Natural Bioactive Products on the Growth and Ginsenoside Contents of Panax ginseng Cultured in an Aeroponic System (United States)

    Kim, Geum-Soog; Lee, Seung-Eun; Noh, Hyung-Jun; Kwon, Hyuck; Lee, Sung-Woo; Kim, Seung-Yu; Kim, Yong-Bum


    This study was conducted to evaluate the effects of natural bioactive products such as Manda enzyme (T1), Yangmyeongwon (T2), effective microorganisms (T3), and Kelpak (T4) on the growth and ginsenoside contents of Panax ginseng cultured in an aeroponic system using a two-layer vertical type of nutrient bath under natural light conditions. The growth of ginseng plants showed specific characteristics according to the positions in which they were cultured due to the difference of light transmittance and temperature in the upper and lower layers during aeroponic culture in a two-layer vertical type of system. The growth of the aerial part of the leaves and stems of ginseng plants cultured in the lower layer (4,000 to 6,000 lx, 23℃ to 26℃) of the nutrient bath was observed to be superior to that of the ginseng plants cultured in the upper layer (12,000 to 15,000 lx, 25℃ to 28℃). The leaf area was significantly larger in the treatment of T2 and T4 (46.70 cm2) than with other treatments. Conversely, the values of the root weight and root diameter were higher in ginseng plants cultured in the upper layer of the nutrient bath. The root weight was significantly heavier in the treatment of T4 (6.46 g) and T3 (6.26 g) than with other treatments. The total ginsenoside content in the leaves and roots was highest in the ginseng plants cultured by the treatment of T1, at 16.20%, while the total ginsenoside content obtained by other treatments decreased in the order of T4, T5 (control), T2, and T3, at 13.21%, 12.30%, 14.84%, and 14.86%, respectively. The total ginsenoside content of the ginseng leaves was found to be significantly higher in the treatment of T1 in the lower layer of the nutrient bath, at 15.30%, while the content of the ginseng roots in the treatments of T3 and T4, at 1.27% and 1.23%, respectively, was significantly higher than in other treatments in the upper layer of the nutrient bath. PMID:23717147

  10. Mitogenic properties of insulin-like growth factors I and II, insulin-like growth factor binding protein-3 and epidermal growth factor on human breast epithelial cells in primary culture. (United States)

    Strange, Karen S; Wilkinson, Darcy; Emerman, Joanne T


    Insulin-like growth factor-I (IGF-I) and insulin-like growth factor-II (IGF-II) are growth factors implicated in mammary gland development and are believed to be involved in breast cancer. However, the interactions between components of the IGF system and breast epithelial cells, which give rise to breast cancer, are not well understood. We have investigated the mitogenic properties of IGF-I, IGF-II, IGF binding protein-3 (IGFBP-3) and epidermal growth factor (EGF) on human breast epithelial cells (HBEC) in primary culture. We show that, under serum-free conditions, HBEC are stimulated to grow in response to IGF-I and IGF-II in a dose-dependent manner. IGF-I and EGF, a potent stimulator of HBEC growth in primary culture and also associated with breast cancer, appear to stimulate HBEC in a synergistic manner. IGFBP-3 inhibits the stimulation by IGF-I, IGF-II and IGF-I plus EGE In addition, it appears that IGFBP-3 has an inhibitory effect on HBEC growth that is IGF-independent. This study is the first to address the effects of IGF-I, IGF-II and IGFBP-3 alone and in combination with EGF on HBEC growth in primary culture. Characterizing the role of the IGF system in normal breast biology is significant because the system has been implicated in breast cancer and a number of the anti-estrogens used in treatment are believed to function through the IGF system.

  11. The metabolism of neonicotinoid insecticide thiamethoxam by soil enrichment cultures, and the bacterial diversity and plant growth-promoting properties of the cultured isolates. (United States)

    Zhou, Guang-Can; Wang, Ying; Ma, Yuan; Zhai, Shan; Zhou, Ling-Yan; Dai, Yi-Jun; Yuan, Sheng


    A soil enrichment culture (SEC) rapidly degraded 96% of 200 mg L(-1) neonicotinoid insecticide thiamethoxam (TMX) in MSM broth within 30 d; therefore, its metabolic pathway of TMX, bacterial diversity and plant growth-promoting rhizobacteria (PGPR) activities of the cultured isolates were studied. The SEC transformed TMX via the nitro reduction pathway to form nitrso, urea metabolites and via cleavage of the oxadiazine cycle to form a new metabolite, hydroxyl CLO-tri. In addition, 16S rRNA gene-denaturing gradient gel electrophoresis analysis revealed that uncultured rhizobacteria are predominant in the SEC broth and that 77.8% of the identified bacteria belonged to uncultured bacteria. A total of 31 cultured bacterial strains including six genera (Achromobacter, Agromyces, Ensifer, Mesorhizobium, Microbacterium and Pseudoxanthomonas) were isolated from the SEC broth. The 12 strains of Ensifer adhaerens have the ability to degrade TMX. All six selected bacteria showed PGPR activities. E. adhaerens TMX-23 and Agromyces mediolanus TMX-25 produced indole-3-acetic acid, whereas E. adhaerens TMX-23 and Mesorhizobium alhagi TMX-36 are N2-fixing bacteria. The six-isolated microbes were tolerant to 200 mg L(-1) TMX, and the growth of E. adhaerens was significantly enhanced by TMX, whereas that of Achromobacter sp. TMX-5 and Microbacterium sp.TMX-6 were enhanced slightly. The present study will help to explain the fate of TMX in the environment and its microbial degradation mechanism, as well as to facilitate future investigations of the mechanism through which TMX enhances plant vigor.

  12. Establishment of a 2-week canine skin organ culture model and its pharmacological modulation by epidermal growth factor and dexamethasone. (United States)

    Abramo, Francesca; Pirone, Andrea; Lenzi, Carla; Vannozzi, Iacopo; Della Valle, Maria Federica; Miragliotta, Vincenzo


    Although canine skin models are already available as either monocellular or organotypic cultures, they only partly recapitulate normal skin morphological features and function. The objective of this study was to establish a canine serum-free skin organ culture model and verify whether dexamethasone could rescue epidermal growth factor-induced changes. The study of morphological changes as a response to pharmacological substances may indeed help to investigate skin physiology and pathology. Normal skin was obtained from five client-owned dogs subjected to surgical procedures unrelated to dermatological conditions. Two experimental designs were performed: (i) two-week viability of the skin culture; (ii) dexamethasone (DMS) inhibition of epidermal growth factor (EGF)-induced effects. Serum-free submerged organ cultures were established in Williams' E medium supplemented with penicillin-streptomycin, insulin, hydrocortisone and l-glutamine. General morphological features of skin anatomical structures were well maintained up to day 14, scattered pyknotic nuclei were visible in the epidermis from day 7. Normal keratinocyte differentiation was confirmed by cytokeratin (K) 10, K14 and loricrin immunostaining. Epidermal thickness did not decrease throughout the study. A decrease in keratinocyte proliferation was observed at day 7 and 14. Treatment with EGF induced both keratinocyte proliferation and thickening of the epidermis; both responses were counteracted by DMS. Treatment with EGF increased the length of epithelial tongues at the edge of the skin explants; this effect was further enhanced by DMS supplementation. Our findings demonstrate the potential use of a full-thickness canine skin organ culture model for the study of skin physiology and pharmacological response to exogenous compounds, especially in the field of re-epithelialisation and keratinization disorders.

  13. Growth performance and proximate composition of mixed cultures of marine microalgae (Nannochloropsis sp. & Tetraselmis sp. with monocultures

    Directory of Open Access Journals (Sweden)

    Wasana Arkronrat


    Full Text Available The growth performance and proximate composition of two marine microalgae, Nannochloropsis sp. and Tetraselmis sp., in monoculture and mixed culture of both species under laboratory conditions were compared. Microalgae were cultured in enriched seawater (28 ppt in 8 L plastic bottles using Conway medium. Microalgae were grown in a temperature-controlled room (25°C under continuous illumination at a light intensity of about 3,000 Lux for 240 hours (10 days. Results showed that maximum cell density and growth rate of both species (45.06±2.67×105 cells ml-1 and 0.43±0.02 day-1, reps. in mixed culture had a significantly higher than Tetraselmis sp. (5.09±0.99×105 cells ml-1 and 0.16±0.03 day-1, reps., but not significantly different with Nannochloropsis sp. (29.25±22.70×105 cells ml-1 and 0.37±0.14 day-1, reps. in monoculture. For proximate composition, protein content of both species in mixed culture and Nannochloropsis sp. in monoculture was significantly higher than for Tetraselmis sp. in monoculture after 120 and 240 hours of cultivation. Carbohydrate and lipid contents of each treatment were found to be significantly different after 120 hours of cultivation but not significantly different after 240 hours of cultivation. Therefore, this study indicates that an alternative option for commercial microalgal production is to use a mixed culture to obtain higher production.

  14. Influence of Temperature on Growth and Production of Pectenotoxin-2 by a Monoclonal Culture of Dinophysis caudata. (United States)

    Basti, Leila; Uchida, Hajime; Matsushima, Ryoji; Watanabe, Ryuichi; Suzuki, Toshiyuki; Yamatogi, Toshifumi; Nagai, Satoshi


    The effects of temperature on growth and production of Lipophilic Toxins (LT) by a monoclonal culture of Dinophysis caudata was studied. The cell density of D. caudata increased significantly with increasing temperature, and was the highest under 27, 30, and 32.5 °C. Temperature affected the average specific growth rate (µ) during the exponential growth phase (EG), which increased from 15 °C to 30 °C, and then decreased at 32.5 °C. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that this strain of D. caudata produced only pectenotoxin-2 (PTX-2) whose concentration increased significantly with incubation period, except at 32.5 °C. It was significantly different between temperatures ≤18 °C, ≥21 °C, and 32.5 °C. The cellular toxin production (CTP, pg·cell(-1)·day(-1)) showed variation with growth phase and temperature, except at 32.5 °C. The average net toxin production (Rtox) was not affected by temperature. During EG, the average specific toxin production rate (µtox) increased significantly with increase in temperature, reaching a peak of 0.66 ± 0.01 day(-1) at 30 °C, and then decreased. Over the entire growth span, µtox was significantly correlated to µ, and this correlation was most significant at 27 and 30 °C. During EG, µtox was affected by both temperature and growth. This study shows that temperature affects growth and toxin production of this strain of D. caudata during EG. In addition, a positive correlation was found between toxin production and growth.

  15. A cellular automata model for simulating fed-batch penicillin fermentation process

    Institute of Scientific and Technical Information of China (English)

    Yu Naigong; Ruan Xiaogang


    A cellular automata model to simulate penicillin fed-batch fermentation process(CAPFM)was established in this study,based on a morphologically structured dynamic penicillin production model,that is in turn based on the growth mechanism of penicillin producing microorganisms and the characteristics of penicillin fed-batch fermentation.CAPFM uses the three-dimensional cellular automata as a growth space,and a Moore-type neighborhood as the cellular neighborhood.The transition roles of CAPFM are designed based on mechanical and structural kinetic models of penicillin batch-fed fermentation processes.Every cell of CAPFM represents a single or specific number of penicillin producing microorganisms,and has various state.The simulation experimental results show that CAPFM replicates the evolutionary behavior of penicillin batch-fed fermentation processes described by the structured penicillin production kinetic model accordingly.

  16. Automatic endpoint determination for batch tea dryers

    NARCIS (Netherlands)

    Temple, S.J.; Boxtel, van A.J.B.


    A laboratory batch fluid-bed dryer was developed for handling small samples of tea for experimental batch manufacture, and this dryer required a means of stopping drying when the process was complete. A control system was devised which requires only the initial weight of the sample to be entered int

  17. Automatic Endpoint Determination for Batch Tea Dryers

    NARCIS (Netherlands)

    Temple, S.J.; Boxtel, van A.J.B.


    Agricultural Engineering and Physics, Wageningen University, Bomenweg 4, Wageningen, 6703 HD, The Netherlands Abstract: A laboratory batch fluid-bed dryer was developed for handling small samples of tea for experimental batch manufacture, and this dryer required a means of stopping drying when the p

  18. Supervision of Fed-Batch Fermentations

    DEFF Research Database (Denmark)

    Gregersen, Lars; Jørgensen, Sten Bay


    Process faults may be detected on-line using existing measurements based upon modelling that is entirely data driven. A multivariate statistical model is developed and used for fault diagnosis of an industrial fed-batch fermentation process. Data from several (25) batches are used to develop a mo...

  19. Following an Optimal Batch Bioreactor Operations Model

    DEFF Research Database (Denmark)

    Ibarra-Junquera, V.; Jørgensen, Sten Bay; Virgen-Ortíz, J.J.;


    The problem of following an optimal batch operation model for a bioreactor in the presence of uncertainties is studied. The optimal batch bioreactor operation model (OBBOM) refers to the bioreactor trajectory for nominal cultivation to be optimal. A multiple-variable dynamic optimization of fed-b...

  20. Family based dispatching with batch availability

    NARCIS (Netherlands)

    van der Zee, D.J.


    Family based dispatching rules seek to lower set-up frequencies by grouping (batching) similar types of jobs for joint processing. Hence shop flow times may be improved, as less time is spent on set-ups. Motivated by an industrial project we study the control of machines with batch availability, i.e

  1. Norton's theorem for batch routing queueing networks

    NARCIS (Netherlands)

    Bause, Falko; Boucherie, Richard J.; Buchholz, Peter


    This paper shows that the aggregation and decomposition result known as Norton’s theorem for queueing networks can be extended to a general class of batch routing queueing networks with product-form solution that allows for multiple components to simultaneously release and receive (batches of) custo

  2. The synthetic inhibitor of Fibroblast Growth Factor Receptor PD166866 controls negatively the growth of tumor cells in culture

    Directory of Open Access Journals (Sweden)

    Castelli Mauro


    Full Text Available Abstract Background Many experimental data evidence that over-expression of various growth factors cause disorders in cell proliferation. The role of the Fibroblast Growth Factors (FGF in growth control is indisputable: in particular, FGF1 and its tyrosine kinase receptor (FGFR1 act through a very complex network of mechanisms and pathways. In this work we have evaluated the antiproliferative activity effect of PD166866, a synthetic molecule inhibiting the tyrosin kinase action of FGFR1. Methods Cells were routinely grown in Dulbecco Modified Eagle's medium supplemented with newborn serum and a penicillin-streptomycin mixture. Cell viability was evaluated by Mosmann assay and by trypan blue staining. DNA damage was assessed by in situ fluorescent staining with Terminal Deoxynucleotidyl Transferase dUTP nick end labeling (TUNEL assay. Assessment of oxidative stress at membrane level was measured by quantitative analysis of the intra-cellular formation of malonyl-dialdheyde (MDA deriving from the decomposition of poly-unsaturated fatty acids. The expression of Poly-ADP-Ribose-Polymerase (PARP, consequent to DNA fragmentation, was evidenced by immuno-histochemistry utilizing an antibody directed against an N-terminal fragment of the enzyme. Results The bioactivity of the drug was investigated on Hela cells. Cytoxicity was assessed by the Mosmann assay and by vital staining with trypan blue. The target of the molecule is most likely the cell membrane as shown by the significant increase of the intracellular concentration of malonyl-dihaldheyde. The increase of this compound, as a consequence of the treatment with PD166866, is suggestive of membrane lipoperoxidation. The TUNEL assay gave a qualitative, though clear, indication of DNA damage. Furthermore we demonstrate intracellular accumulation of poly-ADP-ribose polymerase I. This enzyme is a sensor of nicks on the DNA strands and this supports the idea that treatment with the drug induces cell

  3. Waste Water Derived Electroactive Microbial Biofilms: Growth, Maintenance, and Basic Characterization



    The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup controlled by a potentiostat. Thereby the use of potentiostats allows an exact adjustment of the electrode potential and ensures reproducible microbial culturing conditions. During growth the current production is monitored using chronoamperometry (CA). Based on these data the maximum current density (j max) and the coulombic efficiency (CE) are d...

  4. Dynamic behavior of Yarrowia lipolytica in response to pH perturbations: dependence of the stress response on the culture mode. (United States)

    Timoumi, Asma; Cléret, Mégane; Bideaux, Carine; Guillouet, Stéphane E; Allouche, Yohan; Molina-Jouve, Carole; Fillaudeau, Luc; Gorret, Nathalie


    Yarrowia lipolytica, a non-conventional yeast with a promising biotechnological potential, is able to undergo metabolic and morphological changes in response to environmental conditions. The effect of pH perturbations of different types (pulses, Heaviside) on the dynamic behavior of Y. lipolytica W29 strain was characterized under two modes of culture: batch and continuous. In batch cultures, different pH (4.5, 5.6 (optimal condition), and 7) were investigated in order to identify the pH inducing a stress response (metabolic and/or morphologic) in Y. lipolytica. Macroscopic behavior (kinetic parameters, yields, viability) of the yeast was slightly affected by pH. However, contrary to the culture at pH 5.6, a filamentous growth was induced in batch experiments at pH 4.5 and 7. Proportions of the filamentous subpopulation reached 84 and 93 % (v/v) under acidic and neutral conditions, respectively. Given the significant impact of neutral pH on morphology, pH perturbations from 5.6 to 7 were subsequently assayed in batch and continuous bioreactors. For both process modes, the growth dynamics remained fundamentally unaltered during exposure to stress. Nevertheless, morphological behavior of the yeast was dependent on the culture mode. Specifically, in batch bioreactors where cells proliferated at their maximum growth rate, mycelia were mainly formed. Whereas, in continuous cultures at controlled growth rates (from 0.03 to 0.20 h(-1)) even closed to the maximum growth rate of the stain (0.24 h(-1)), yeast-like forms predominated. This pointed out differences in the kinetic behavior of filamentous and yeast subpopulations, cell age distribution, and pH adaptive mechanisms between both modes of culture.

  5. Making sense of institutional change in China: The cultural dimension of economic growth and modernization


    Herrmann-Pillath, Carsten


    Building on a new model of institutions proposed by Aoki and the systemic approach to economic civilizations outlined by Kuran, this paper attempts an analysis of the cultural foundations of recent Chinese economic development. I argue that the cultural impact needs to be conceived as a creative process that involves linguistic entities and other public social items in order to provide integrative meaning to economic interactions and identities to different agents involved. I focus on three p...

  6. Consequence Identification for Maloperation in Batch Process

    Institute of Scientific and Technical Information of China (English)

    张玉良; 张贝克; 马昕; 曹柳林; 吴重光


    Batch processes are important in chemical industry, in which operators usually play a major role and hazards may arise by their inadvertent acts. In this paper, based on hazard and operability study and concept of qualitative simulation, an automatic method for adverse consequence identification for potential maloperation is proposed. The qualitative model for production process is expressed by a novel directed graph. Possible operation deviations from normal operating procedure are identified systematically by using a group of guidewords. The pro-posed algorithm is used for qualitative simulation of batch processes to identify the effects of maloperations. The method is illustrated with a simple batch process and a batch reaction process. The results show that batch processes can be simulated qualitatively and hazards can be identified for operating procedures including maloperations. After analysis for possible plant maloperations, some measures can be taken to avoid maloperations or reduce losses re-sulted from maloperations.